Allogeneic dermal progenitor fibroblasts constitute cytotherapeutic contenders for modern cutaneous regenerative medicine. Based on advancements in the relevant scientific, technical, and regulatory fields, translational developments have slowly yet steadily led to the clinical application of such biologicals and derivatives. To set the appropriate general context, the first aim of this study was to provide a current global overview of approved cell and gene therapy products, with an emphasis on cytotherapies for cutaneous application. Notable advances were shown for North America, Europe, Iran, Japan, and Korea. Then, the second and main aim of this study was to perform a retrospective analysis on the various applications of dermal progenitor fibroblasts and derivatives, as clinically used under the Swiss progenitor cell transplantation program for the past three decades. Therein, the focus was set on the extent and versatility of use of the therapies under consideration, their safety parameters, as well as formulation options for topical application. Quantitative and illustrative data were summarized and reported for over 300 patients treated with various cell-based or cell-derived preparations  in Lausanne since 1992. Overall, this study shows the strong current interest in biological-based approaches to cutaneous regenerative medicine from a global developmental perspective, as well as the consolidated local clinical experience gathered with a specific and safe allogeneic cytotherapeutic approach. Taken together, these current and historical elements may serve as tangible working bases for the further optimization of local and modern translational pathways for the provision of topical cytotherapeutic care. Topical cytotherapies designed for cutaneous reconstructive surgery or regenerative medicine protocols were among the first to be developed and clinically applied on large scales in the late 20th century. The pioneer works of Reinwald and Green on cultured autologous keratinocytes for the management of burn wounds constitute notable examples ,2. DurinWhile generally beneficial effects have resulted from the application of recent quality norms  and regulatory requirements in the cell therapy field, several constrictive elements have often limited new technologies’ and applications’ progression from the scientific proof-of-concept state to phase I/II clinical trials ,7,8,9. TThe previously exposed factors and drivers that have shaped the global cell therapy ecosystem have also applied to the clinical use of allogeneic dermal progenitor fibroblasts, which notably constitute cytotherapeutic contenders in Swiss cutaneous regenerative medicine ,16. BaseThe overarching goal of this study was to present three decades of acquired clinical hindsight and current perspectives on the use of progenitor dermal fibroblasts and derivatives in Swiss cutaneous regenerative medicine. The retrospective analysis was based on selected internal factors  and several external factors  in the interventions under consideration. To set the appropriate context, the first aim of this study was to provide a global overview of approved cell and gene therapy products, with an emphasis on cytotherapies for cutaneous application. Then, to provide a specific point of discussion, the second and main aim of this study was to perform a retrospective analysis on the various applications of dermal progenitor fibroblasts and derivatives, as clinically used under the Swiss progenitor cell transplantation program for three decades. Therein, the focus was set on the extent and versatility of use of the therapies under consideration, their safety parameters, as well as formulation options for simple topical application. Overall, this study firstly demonstrates the strong current interest in biological-based approaches to cutaneous regenerative medicine from a global developmental perspective. Secondly, the subject is discussed more deeply around the example of dermal progenitor cells for clinical use, where important consolidated local clinical experience has been gathered with a safe and versatile allogeneic cytotherapeutic approach.In order to provide an updated and summarized overview of the global cytotherapeutic ecosystem , multiple sources were cross-referenced. First, several recent scientific literature reviews dealing with regulatory processing and market approvals for cell and gene therapy products were compiled. Then, the relevant national or supra-national online public registries were consulted for the retained products or therapies and for newly approved or retracted products. Finally, the relevant websites of the various product manufacturers or clinical sponsors were reviewed. In cases where conflicting information between available sources was collected, the contents of the official public healthcare agency registries were retained. Data presentation for this part of the study was performed in graphical form, with listing both of the various product classes and of the individual products. The geographical distribution of the various countries of approval for given products and therapies was included as well.The acquisition of previously reported studies  and of the original unpublished data for the present work was performed by the systematic and comprehensive compilation of research, manufacturing, regulatory, and clinical files generated under the Swiss progenitor cell transplantation program since 1991. In particular, the necessary documentary elements were retrieved appropriately from the archives of the Plastic and Reconstructive Surgery Service (CPR) and the Burn Center of the CHUV . In detail, the general information, specificities, and data related to cell therapy manufacturing and to related clinical work were mainly synthesized from available regulatory documentation , from manufacturing records, or from treated patient files. With regards to primary patient data, appropriate information anonymization and data codification or security protocols were used at all times during the study.To set the appropriate general context for the present work, a summarized global overview of currently approved cell and gene therapy products was prepared, with an emphasis on cytotherapies for cutaneous application. A condensed worldwide overview of the currently approved advanced therapy medicinal products (ATMP), distributed into product sub-categories and classified by therapeutic indication, is presented in Importantly, a consideration of global ATMP approval statistics reveals that, for cell therapy medicinal products (CTMP), the most important therapeutic indication with regards to product or therapy numbers are “skin and soft tissue disorders” , the performed cytotherapy ecosystem analysis also took into account some historical elements which predated the ATMP nomenclature and regulatory classification system. Therein, many landmarks and pioneer preparations based on autologous or allogeneic skin cells have been marketed under an array of FDA procedures , PMA, orphan drug, etc.) ,21. WhilIn this domain, distinctions are made between acellular and cellular skin substitutes designed for cutaneous wound healing promotion. While acellular products  may be effectively used as early and temporary coverage solutions in burn centers, deep and/or extensive burns often require the use of specific cytotherapies ,22. The In addition to the developments of novel cell carriers and in vitro biological material processing techniques, it is worth mentioning alternatives to classical split-thickness skin biopsy harvest techniques , such as those performed in fractional epidermal grafting or epidermal blister grafting ,24. SuchOverall consideration of the presented data and specificities on the global cytotherapeutic ecosystem indicates that topical cell-based preparations have always been and continue to be key drivers in recent scientific, technical, regulatory, and commercial developments. Specifically, it was shown that, despite the current existence of well-defined product or therapy classifications and related regulatory guidelines, most of the pioneer registration work in the topical cytotherapy field dates back to the 1990s and early 2000s . Such elAs previously mentioned, a restricted number of countries or regions  have historically been implicated in the development and clinical use of cytotherapies for cutaneous application B. Aside Such undertakings were motivated by the high unmet needs of severe burn victims, as well as the high therapeutic potential and technical simplicity of primary progenitor dermal fibroblast cell banking. Specifically, a main component of this converging evolution in therapeutic approaches has been the general homogenization of quality standards for cell therapy manufacture and use, starting with good manufacturing practice (GMP) requirements. Therefore, several groups around the globe have made tangible progress around the use of allogeneic dermal progenitor cells up to the clinical phase for burns and donor site wounds, as summarized hereafter in several relevant randomized clinical trial examples .Firstly, the Tehran University of Medical Sciences  has recently completed a phase I clinical trial  for burn patient donor-site wounds (DSW) using prenatal-derived cellular materials . ThereinBoth of the mentioned clinical trial examples  are similar to the clinical work performed under the Swiss progenitor cell transplantation program, albeit with several technical and clinical adaptations ,26. SpecOverall, the fact that multiple clinical groups around the world have been investigating similar dermal progenitor cell sources for a certain number of years  points toward the consolidated safety aspects of the interventions under consideration. Indeed, when reviewing the available information about the clinical work mentioned, no treatment-related adverse outcome or event has to date warranted a study interruption for safety reasons ,32,33,34At a time when cell therapies were prescribed and administered to patients as magistral preparations in Switzerland , much leeway was leveraged for the translational development and clinical implementation of novel topical cytotherapeutics ,30. SuchIn order to provide specific elements on the historical extent and versatility of use for the considered therapies, their safety parameters, as well as formulation options for topical application, summarized data were prepared. Firstly, important methodological elements regarding starting biological material obtention and processing for progenitor cell banking, as well as cytotherapeutic product  manufacture, are presented as supporting information in Firstly, summarized patient statistics  for topical treatment using off-the-shelf semi-solid preparations and progenitor cell derivatives were presented hereafter as classified by initial clinical presentation: eczema (5 patients), actopic dermatitis (28 patients), atrophie blanche (3 patients), burns (7 patients), post-skin grafting in burn patients (8 patients), post-skin grafting in ulcer patients (13 patients), scar management (4 patients), severely crevassed and chapped hands (10 patients), radiodermatitis (2 patients), psoriasis (3 patients), rosacea (2 patients), scars and keloids (12 patients), vestibulitis (2 patients), vulvar-vestibulitis (23 patients), vulvar lichen sclerosis (9 patients), atopic dermatitis and dryness (4 patients) ,32. TreaSecondly and as concerns the use of viable dermal progenitor fibroblasts  used in combination with collagen scaffolds to constitute progenitor biological bandages, a detailed summary of the cutaneous clinical indications and of the cytotherapeutic product forms used to treat patients under the Swiss progenitor cell transplantation program is presented in Additional details on the various clinical studies performed in Lausanne around the use of the considered progenitor biological bandages are listed in As concerns the therapeutic use of PBB constructs for the management of refractory lower-leg ulcers, the results of a phase I clinical study have shown that patients tolerated multiple treatments, displayed no adverse effects, and ulcers were observed to undergo repair processes similar to those seen in 3rd degree burn victims  34]. Im. Im34]. An illustrated summary of the evolutive numbers of clinical cytotherapeutic units  manufactured for clinical use in Lausanne under the Swiss progenitor cell transplantation program is presented in Building on this in-house experience and on successive regulatory approvals  for further investigational allogeneic cytotherapeutic use, multiple studies and standardized clinical trials have been or are being conducted around the progenitor cells under consideration . In LausOverall, the available records show that over 200 patients have been treated with the progenitor biological bandages under consideration for a variety of clinical indications under the Swiss progenitor cell transplantation program . TreatedOverall consideration of the available clinical treatment statistics for dermal progenitor cells and derivatives in Lausanne has outlined the safety and the utility of such highly specialized care provision approaches for >300 patients, all indications and treatment modalities combined. The integration of these three decades of safe clinical experience and the constitution of a wide body of knowledge has enabled us to draw robust conclusions on the retained approach to topical cytotherapeutic care. Parallelly, the historical undertakings summarized herein have contributed to placing primary dermal progenitor cells at the forefront of allogeneic cytotherapies elaborated in Switzerland.Consideration of the data reported herein notably enables an assessment on the high versatility of banked primary progenitor fibroblasts and derivatives for topical therapeutic application in cutaneous regenerative medicine. Versatility was considered firstly from a technical applicability standpoint, as it was demonstrated that both the biological materials and the final product formulations could be easily adapted in view of clinical use. Specifically, both proliferation-capable cells and growth-arrested cells  could be used in combination with collagen scaffolds to constitute PBBs or PBIs for clinical use, with no reported differences in safety outcomes  has evidenced a trend of reduced needs for corrective interventions or for subsequent skin grafting, as well as significantly reduced hypertrophic scarring  . Thi. Thi5]. When considering the influence of regulatory bodies on the global cytotherapy ecosystem, many hurdles have been identified in the European or North American regulatory ecosystems pertaining to translational development of biologicals  ,8,9,10. A consistent technical limitation has been evidenced around the use of extemporaneously reconstituted PBBs, namely the cumbersome and costly cold chain maintenance for cryogenic storage and shipping . Indeed,https://www.neocutis.com/, accessed on 9 September 2022) over the past decades and attaining global reach [A key parameter in ensuring clinical success in topical cytotherapeutic care may reside in the use of multiple small product doses administered regularly over an extended time-period, for production of enhanced results as compared to few large doses. Based on this concept and on preliminary in-house clinical observations, the considered progenitor cell extracts were formulated into semi-solid topical formulations for wound repair promotion ,32. Furtal reach ,18. BaseAs regards the endpoints and readouts of cutaneous clinical care and patient follow-up, specific parameters have been identified by the authors, for the tangible and optimal assessment of the effects of the cytotherapeutics and derivatives under consideration. As mentioned, appropriate follow-up and maintenance treatments are vital in ensuring appropriate cutaneous wound healing, including the elimination of aggravating factors , if possible. Furthermore, temporal and qualitative healing parameters should be defined with great care, as clinical investigators should focus specifically on the overall skin quality and functionality following repair and on patient quality of life following the cytotherapeutic interventions . OverallThe extensive clinical experience set forth herein has enabled a constant challenging of the boundaries of cutaneous regenerative medicine care provision, with the development of adapted solutions and tools. Therefore, recent research directions under the Swiss progenitor cell transplantation program have comprised the development of ancillary components or processing methods, aiming mainly to fight infectious risks in burn patient populations, as well as to provide improved, stable, and safe cytotherapy-inspired derivative products. Key considerations shaping and orienting the next steps in therapy development have been cell source processing, clinical administration modalities, and the use of cell-derived cell-free biological materials, to cite only a few recent scientific and technical areas of focus ,53,54,55First-generation PBBs: Collagen scaffolds seeded with viable and growth-capable dermal progenitor fibroblasts  ,16.Second-generation PBBs (PBI): Collagen scaffolds seeded with viable and growth-arrested  dermal progenitor fibroblasts .Third-generation PBBs: Similar to the first generation, with addition of antimicrobial dendrimers, for combination of intended effects and management of the infectious risk  .Fourth-generation PBBs: Appropriate vehicle yielding temperature-stabilized non-viable dermal progenitor fibroblasts, for an off-the-shelf availability  .Fifth-generation PBBs: Appropriate vehicle yielding cell-derived cell-free and temperature-stabilized therapeutic extracts, for an off-the-shelf availability .For the optimal illustration of the adopted past and present academic and parallel research directions in Lausanne, the various generations of investigated progenitor biological bandages are summarily listed hereafter, along with the corresponding stages of research or development and clinical use:Overall, the development efforts mentioned hereabove have been deployed with the objectives of maximizing the scope of potential therapeutic gains procured by the intervention, augmenting the availability of treatments, and enhancing manufacturing quality . While sIn particular, the results of previous in vitro studies and preclinical or clinical applications have suggested that the retention of cellular viability or the presence of the original cellular structures was not required to conserve specific therapeutic functions ,66,67,68Building on the existence of multi-centric GMP manufacturing capacities of the considered dermal progenitor cell sources and on the previous validation by multiple regulatory agencies for clinical investigation of the corresponding cytotherapies, methodological updates are currently being implemented into the clinical work performed in the Lausanne burn center . FollowiIn detail, cantonal ethical validation has been granted  for the mentioned upcoming clinical trial on PBBs in the context of the CHUV Priority Project Bru_PBB. This phase I/II interventional, prospective, and randomized monocentric clinical study  will include at least 76 burn patients over the next five to ten years in two study arms . The objectives of this new clinical study, in addition to the authorized investigational use of PBBs, comprise the potential renewed demonstration of short-term and long-term cytotherapeutic care efficacy .Overall, the presented case-study of PBBs is useful for the demonstration that with proper methodological devising and technical adaptation, specific cytotherapeutic interventions may evolve and persist despite drastic changes in the local healthcare ecosystem. Therein, process and therapy versatility may be usefully combined with the acquired hindsight and analysis of current global regulatory trends, to ensure that continued focus and driving forces locally animate the forefront of topical cytotherapeutic care.The aim of this study was to set forth the consolidated clinical data, experience, and hindsight gathered over thirty years around the use of allogeneic dermal progenitor fibroblasts and derivatives for topical therapeutic applications. Such practices were set in the context of the current global cytotherapy ecosystem. Original data was provided for various clinical steps and issues addressed under the Swiss progenitor cell transplantation program since 1991 for cutaneous cytotherapeutic care provision. Along with the high versatility and robustness of primary progenitor fibroblasts used as biological starting materials, critical aspects of clinical safety were summarized herein for such allogeneic cells . Three decades of clinical work, with the needs of over 300 patients addressed, have generated robust hindsight and technical know-how for cell-based and cell-derived therapy translation and transposition. Overall, this study covered the strong current and global interest in biological-based approaches to cutaneous regenerative medicine, with an orientation toward the clinical use of allogeneic cytotherapies based on dermal progenitor fibroblasts. Taken together, these current and historical elements may serve as tangible working bases for further optimization of local and modern translational pathways, for the provision of high-quality topical cytotherapeutic care.


To study the role of bacterial biofilm (BBF) in the formation of chronic osteomyelitis and its prevention and treatment.  In this paper, a large amount of relevant literature was searched for analysis and summary, and the key words “chronic osteomyelitis,” “bacterial biofilm,” “infection,” and “debridement” were searched in databases, mainly CNKI, Wanfang, and Wipu. The search was conducted until December 2020. The role of bacterial biofilm formation in chronic osteomyelitis and its prevention were analyzed.  Chronic osteomyelitis is formed mainly due to poor blood supply and drug-resistant bacteria, of which cellular biofilm is the most important cause. BBF forms on the surface of necrotic soft tissue and bone tissue, which has a protective effect on bacteria and greatly enhances their resistance to antibiotics, leading to difficulties in complete bacterial clearance and recurrent infections in osteomyelitis.  Through an in-depth study of the molecular biology and signal transduction of osteomyelitis biofilm, antibiotic biofilm treatment strategies and surgical debridement remain the focus of clinical translation of chronic osteomyelitis. Most osteomyelitis is intraoperative vegetative osteomyelitis and open fractures with an incidence of 80%, of which about 30% develop chronic osteomyelitis . In receOsteonecrosis, sclerosis, and fistulas are common clinical signs of chronic osteomyelitis and patients are at risk of decay and recurrence of infection . In addiThe concept of biofilm was first introduced in 1978 . The comOsteomyelitis due to biofilm infection is mostly subacute or chronic and usually occurs 3-10 weeks after trauma or surgery; bacteria in biofilm of osteomyelitis originate from planktonic or endophytic bacteria on the surface of open wound contaminants . As a reStaphylococcus. There is a certain correlation between infection and soft tissue damage. Bacteria are often not enough to cause osteomyelitis. Therefore, the treatment of chronic osteomyelitis should focus on soft tissues. Acute infection is difficult to diagnose at an early stage, but early and correct antibiotic therapy can eliminate acute bone marrow infection in about 4 weeks, and the likelihood of acute infection developing into chronic osteomyelitis is high.Bacterial infection can destroy bone and cause chronic inflammation, the clinical symptoms of which are ulcers and rot in the limbs, swelling, and pigmentation in the skin tissue. Chronic osteomyelitis occurs mostly in the tibia and femur of young men. The most common cause is fracture and multiple surgeries. It often occurs repeatedly and is difficult to cure. The most common pathogenic bacteria are Gram-positive bacteria, Gram-negative bacteria, and Early diagnosis and treatment of chronic osteomyelitis is the primary key which can usually be detected by appropriate imaging or laboratory measurements. Chronic osteomyelitis can be diagnosed if the number of white blood cells increases in routine blood tests. Imaging can be done by X-ray, CT, MRI, and radionuclide examination. Radionuclide examination combined with CT has a high diagnostic value in the diagnosis of early osteomyelitis. X-ray can detect a periosteal reaction, osteosclerosis, and sinus canal, but it can be found only 2 weeks after bone destruction. It has low sensitivity in the early stage of chronic osteomyelitis and cannot distinguish fracture from chronic osteomyelitis. Ultrasound can detect signs of osteomyelitis at an early stage such as soft tissue swelling, periosteal thickening, and effusion. It also has certain advantages of directing needle subperiosteal injection, low cost, low radiation, and easy to accept patients. Compared to X-ray imaging, CT can show periosteal reaction, bone destruction, and necrosis around the focus in more detail. MRI is better than X-ray and CT diagnosis of chronic osteomyelitis. It can be detected at 3 to 5 days of osteomyelitis and has a sensitivity and specificity of more than 90% , 24.α MRI imaging system with 2 receptor-labeled probes can distinguish between aseptic and infectious inflammation, but this technique has not been applied clinically since S. aureus is the most important pathogen of chronic osteomyelitis in clinical practice. Raman spectroscopy has been reported for the diagnosis of chronic osteomyelitis caused by S. aureus, which provides an opportunity to develop an early and rapid diagnosis of this disease; this diagnosis provides an opportunity to develop a diagnostic method for early and rapid diagnosis of this disease [It is increasingly used in clinical practice because it can better differentiate between the bone and surrounding soft tissue and muscle infections, soft tissue, fascia and conjunctiva providing more information to inform the development of surgical plans. However, this technique has some drawbacks: image quality is easily affected by internal implants produced by the object; incorrect diagnosis is due to its oversensitivity to tissue swelling and scar tissue; PET-CT is indicated for osteomyelitis associated with internal fixation –27. It c disease , 29. TheBacterial biofilm contains bacteria, a large amount of water, macromolecular polymers, metabolites, and bacterial lysates. The formation of a multicellular structure is a dynamic process involving the adhesion, development, and maturation of bacteria. Bacterial biofilm formation is associated with many factors such as bacterial species, surrounding environment, surface components of adhesion vectors, and expression of essential genes. Bacterial biofilm has different physiological and biochemical properties at each stage.Bacterial biofilm formation first requires bacteria to stick to the surface of living organisms or plants. The adhesin between bacteria and plants is not specific, whereas the adhesin protein of living organisms requires specific adhesin proteins on the surface of bacteria to identify host surface receptors that are selective and specific. Infection of bacteria is usually divided into two stages: docking and locking. The first case must be the opposite, because bacteria can accidentally come in contact with the adhesive or be driven by the chemical agent. When the distance between the two is less than 1 nm, bacteria are transmitted to the carrier by various forces such as hydrophobic effect, electrostatic effect, van der Waals force, and sterile resistance . BacteriStaphylococcus epidermidis to polyethylene is significantly increased in the presence of surfactant platelets, while adhesion is weakened in the presence of plasma proteins. The bacterial adhesion lock is irreversible from time to time, and when the bacteria dock on the carrier, they secrete polysaccharides which transmit a close combination of the bacteria and the carrier. Different bacteria can secrete different polysaccharides, and one bacterium can secrete different polysaccharides; for example, Staphylococcus epidermidis can secrete polysaccharide adhesin between cells. Through this adhesion, the same or different types of bacteria can also be combined, and the adhesion of one bacterium can improve the adhesion of another bacterium.It is the main force of bacterial transmission. It should be noted that the hydrophobic effect of the carrier may be influenced by other factors such as the adhesion of When the bacteria stick to the surface of the carrier, the bacterial biofilm begins to develop until it matures. First, the expression of bacterial genes changes, and a large number of extracellular polysaccharides are excreted during growth and reproduction. External polysaccharides can bind one or several bacteria to form bacterial clusters, and a large number of microcolonies thicken the biofilm of the bacteria. The amount of nutrients produced by the environment and the smooth transport of metabolic products directly affect the maturity of the bacterial biofilm. In addition, the pH value, osmotic pressure, and oxygen content of bacteria play an important role in the maturation of bacterial biofilm. At this stage, changes in antibiotic resistance, ultraviolet radiation resistance, and the effectiveness of gene exchange are often observed.The structure of mature bacterial biofilm is heterogeneous. Bacteria near the surface of the carrier grow slowly due to lack of nutrients, oxygen, and accumulation of metabolites. For example, the oxygen content of the outer layer of the bacterial biofilm can be 30 times higher than the middle; bacteria on the surface of the grain mill grow actively and metabolize extensively, and when the number of bacteria reaches a certain level, they are released from the bacterial biofilm and form planktonic bacteria . BacteriThe EPS secreted by bacteria can realize the biomembrane barrier effect on antibiotics and improve antibiotic drug resistance through dormant bacteria. The number required for bacterial infection can be reduced through the biofilm barrier, the entry of antibiotics, and the role of sleeping bacteria. Some researchers found that the minimum inhibitory biofilm of the same type of bacteria was 1000 times the minimum inhibitory concentration. However, in vitro experiments have formed a similar infection state; when there is no biofilm on the inner surface of the plant, 104 times more bacteria are required than biofilm, see β-EPS-protected biofilm, bacterial efflux enzymes can maintain concentration and activity [γ-hemolysin AB bacterial biofilm leads to the release of a network of neutrophils, but the network accelerates and thickens biofilm formation [The presence of biofilm can significantly reduce the penetration of antibiotics such as vancomycin and teicoplanin. The principle of special barrier can be divided into the following categories: (1) hydrophobic structure of the biofilm: the main component of the extracellular polymer of the biofilm is water, but the extracellular polymer is not hydrophilic. An extracellular polymer that converts free water and gathers together through polysaccharides, proteins, and other components. The molecular weight of the water mass increases with the maturity of the biofilm. The properties of the water mass are also changing. The water mass with a high molecular weight is usually hydrophobic, and the hydrophobic macromolecular mass is usually formed a hydrophobic film on the surface of the biofilm. It is stable in body fluids. Adding heavy metal ions destroys macromolecular water mass and eliminates hydrophobicity, and the biofilm further decomposes. Low-molecular-weight water is usually hydrophilic in biofilms. This internal hydrophilic and external hydrophobic amphoteric structure complicates the penetration of most hydrophilic antibiotics released from the kidneys and is unable to achieve an effective concentration in the biofilm. (2) Negative charge and acidic environment of the biofilm: because the extracellular polymer of the biofilm and the cell wall of bacteria contain a large amount of polysaccharide and wall phosphate, the biofilm is negatively charged, resulting in positively charged molecules such as aminoglycoside antibiotics and combine with EPS by electrostatic action to prevent further penetration of antibiotics. Experimental results indicate that the difference in pH between the surface and inside of the biofilm may be greater than 1 which may be due to accumulation of H absorbed by the negative charge or accumulation of acid metabolites of bacteria. A significant pH gradient inside and outside the biofilm directly affects antibiotic activity. (3) Biofilm directly inactivates antibiotics: in addition to structural proteins, biofilms also contain functional proteins such as protein secreted by bacteria. In the activity . In vitractivity . As a ceormation . This poStaphylococcus aureus is a restricted immune system based mainly on biofilm. Experiments show that there is an asymmetric relationship between biofilm and host immunity, that is, biofilm can trigger a corresponding immune response, but the immune response cannot effectively remove biofilm. From the point of view of biofilm structure, EPS biofilm is closely bound to bacteria, and bacteria secrete macrophage-permeable biofilm α-toxin and Leukocidin AB, and disappeared from immune monocytosis. Pseudomonas aeruginosa PSL polysaccharide can also protect bacteria from conditional effects. Proteins in biofilms such as serine protease and cysteine protease can break down immunoglobulins and antimicrobial peptides that are secreted by the immune system. From a molecular biology point of view, biofilms pass through interleukin-12 and interleukin-1 β, CCL5, and other cytokines and chemokines attract bone marrow-derived suppressor cells affect macrophage polarization, transfer congenital immunity, promote inflammation, sterilization (type M1), inflammation, and fibrosis immune response (type M2) leading to local immunosuppression, excessive fibrosis, and scarring.Biofilm can resist host-specific and nonspecific immunity and plays an important role in bacterial immunity. Studies have shown that S. aureus can penetrate the vascular endothelium. Therefore, osteoblasts can lead to apoptosis. Biofilms also have the ability to absorb directly into the bone, which can destroy the bone without immunity and osteoclasts in vitro. Gram-negative bacteria can also produce endotoxin and trigger an immune response. Bacteria can change the composition of EPS polysaccharide in the biofilm to control the formation and diffusion of the biofilm to meet the needs of the biofilm at different times.The clinical symptoms of chronic osteomyelitis are mainly due to the release of planktonic bacteria after biofilm maturation . The sepS. aureus, sufficiently inducing phenotypic changes in bacterial biofilms. Biofilm EPS can also bind to plasmids to promote horizontal transfer of resistance genetic information among bacteria and accelerate the transition from drug-resistant to drug-resistant bacteria. Bacteria are better able to adapt to changes in the external environment than individual planktonic bacteria by establishing biofilms to communicate, coordinate, and cooperate among bacteria, see In the biofilm bacterial proliferation process, secondary metabolites are formed into signal molecules. When signal molecules reach the threshold level, they can bind to bacterial receptors and regulate the transcription of target genes so that bacteria can play an important physiological role as a whole at the multicellular level. This regulatory framework is called quorum recognition (QS). In Gram-negative bacteria, the signal molecule is acyl hypericin lactone (AHL). In Gram-positive bacteria, the signal molecule is a self-inducing peptide, and different bacteria can also exchange the self-inducing factor-2 (AI-2), which is a byproduct of bacteria's metabolic methyl cycle. Most bacteria contain two or more groups of quarium detection systems. Changes in the expression profile of bacterial genes in biofilm are regulated by a regulatory network consisting of several quarium detection systems. For example, the LuxS/AI-2 population detection system regulates bacterial polysaccharide expression via the kdpde binary signaling system, while the age population detection system regulates virulence factors in Staphylococcus, including Staphylococcus aureus and Staphylococcus epidermidis. When Staphylococcus biofilm grows in the dead bone, scar, and other tissues, it leads to repeated attacks of infection. Staphylococcus can cause serious infections when the epidermis and mucous membrane are damaged. It can attach to the surface of inactive tissues and medicinal plants, secrete polysaccharides, form a biofilm, and cause infection. Recent studies have confirmed that this polysaccharide component is an intracellular lipopolysaccharide binder. The synthesis of intracellular lipopolysaccharide binder (PIA) depends on the activity of the icaabcd gene, which encodes an intracellular lipopolysaccharide binder synthetase, and its activity is regulated by genes such as ICAR, sigB, and RsbU, while the external environment such as glucose, ethanol, osmolality, temperature, and antibiotics can affect the above regulatory genes. For example, glucose can induce the expression of icaabcd, which is partially dependent on sigB and RsbU genes. RsbU genes are activators of sigB genes, and these two genes can induce biofilm formation. icar genes encode inhibitors of icaabcd genes and ethanol and NaCl can inhibit the expression of icar genes and activate sigB and RsbU genes and induce biofilm formation. However, studies have shown that 30% Staphylococcus epidermidis is intracellular lipopolysaccharide adhesin negative. Another study confirmed that only 1 in 3 Staphylococcus epidermidis isolated from the surface of infected hip prostheses was ICA gene positive. The literature [Staphylococcus isolated from infected pseudopodia did not reduce biofilm formation. The literature [Staphylococcus aureus was not associated with biofilm formation, but intracellular lipopolysaccharide of methicillin-resistant Staphylococcus aureus was associated with biofilm formation. Therefore, it can be concluded that the formation of staphylococcal biofilms caused by intracellular lipopolysaccharide is related to the strain and the surrounding environment. Without the role of intracellular lipopolysaccharide, protein adhesins play an important role in the formation of biofilms. The literature [Staphylococcus epidermidis. However, the mechanism of staphylococcal biofilm formation in the absence of intracellular lipopolysaccharide duplexes needs further investigation.The main pathogenic bacteria of chronic osteomyelitis are terature  found thterature  also fouterature  identifiS. aureus. atlE mutation results in reduced release of bacterial DNA and reduced biofilm formation.The literature  confirmsMany factors including regulatory proteins are involved in the formation of staphylococcal biofilm, and its mechanism needs to be further investigated.The inability of antibiotics to completely remove bacterial biofilms is mainly due to the fact that bacterial biofilms target mainly planktonic bacteria. In recent years, most researchers have studied the relationship between staphylococci and biofilms, mainly focusing on (1) reducing the synthesis of intracellular lipopolysaccharide adhesins and affecting their adhesion and (2) some researchers found that dspb can effectively remove intracellular lipopolysaccharide adhesins and prevent bacterial transmission. However, not all bacteria are completely positive. Some researchers have suggested that bacterial biofilms can inhibit growth through RNA inhibitory peptides and confirmed that the consistency of density detection systems can facilitate the purification of bacterial biofilms from antibiotics . HoweverIn conclusion, in recent years, the treatment of osteomyelitis has changed from previous debridement to debridement, masquelet bone reconstruction or osteotomy, ilizarov bone handling, or negative pressure coagulation. The therapeutic effect has been significantly improved, but the number of surgeries, the length of the treatment process, the secondary trauma, the poor adaptability of patients, and the presence of biofilm make osteomyelitis infection very persistent. It is difficult to resolve with surgical techniques and antibiotics alone. Studying the role of biofilms in osteomyelitis and determining the mechanisms and pathways of biofilm action in osteomyelitis is a trend for future research. Antibiotic biofilm therapy and surgical debridement are hot spots for clinical changes in chronic osteomyelitis. For example, antibiotic transport materials are used to achieve effective concentrations in biofilms or to form a coating on the inner surface of the plant to prevent biofilm adhesion. Interfering with the detection of bacterial populations leading to self-degradation of biofilms; adding antibiotic adjuvants to dormant bacteria to remove bacterial residues along with conventional antibiotics; modulating immune activity to prevent immune polarization; adding Edna or polysaccharidases or phagocytic enzymes to break down EPS components and networks in biofilms to break down and release them. The research and treatment of chronic osteomyelitis has been enhanced by in-depth molecular biology studies and osteomyelitis signaling.


In semi-cold, moderate and warm climates, daughters of French sires had the highest least-square means of LNI index. In cold climate, daughters of German sires had the highest least-square means.We evaluated the impact of imported semen of Holstein bulls from different countries on their daughters’ economic traits using the Lifetime Net Income (LNI) index in different climates of Iran. The first lactation records were collected from 10 big dairy farms in various regions of Iran. Studied traits included milk, fat and protein yields, calving age and calving interval. There were significant differences between sire groups for studied traits in each climate (p < 0.01). The obtained results showed that the highest least-square means of LNI index in semi-cold, moderate and warm climates belonged to the daughters of French sires; however, daughters of German sires were estimated to have the highest least-square means in the cold climate.The aim of the present study was to evaluate the effects of imported semen of Holstein bulls from different countries on the economic traits of their daughters using the Lifetime Net Income (LNI) index in various climates of Iran. The data included the first lactation records of 274,057 Holstein cows collected during 1993 to 2017 by the Animal Breeding Center of Iran from 10 large dairy farms located in various provinces of Iran. The investigated traits included milk, fat and protein yields, calving age and calving interval. Breeding values of progenies were predicted by the Best Linear Unbiased Prediction (BLUP) method under the multi-trait animal model using DMU software. The genetic-economic merit of the progenies was estimated by the LNI index. There were significant differences between the estimated breeding values (EBVs) of sire groups (based on bull semen origin) for milk, fat and protein yields, calving age and calving interval in each climate ( For many years, dairy cattle breeding programs have mainly been directed toward the improvement of production traits. This has been true for most countries, except for Scandinavian countries, whose selection indices have also included health and reproduction, and North American countries, whose selection indices have included conformation traits . On the Selection index is a theory for maximizing genetic progress to achieve maximized profitability . Since 1In the Holstein cattle population, genetic improvement through the selection of males compared to females is of crucial importance . ImportiData used in the present study included the first lactation records of 274,057 Holstein cows from 1993 to 2017 provided by the Animal Breeding Center of Iran (ABCI). The origin of sires was from the United States, Canada, France, the Netherlands, Italy and Germany. The investigated traits included milk, fat and protein yields, calving age and calving interval. Cows with less than 1000 kg and more than 15,000 kg of total milk production, herds with fewer than 30 records and first calving age less than 18 months and more than 42 months were excluded from data.Records were classified under cold, semi-cold, moderate and warm climates based on the climatic data for stations where data collected . Accordiith trait; ith trait ; ith trait; ith trait; and Breeding values of progenies were estimated by the Best Linear Unbiased Prediction (BLUP) methodology under multi-trait animal models using DMU software . The mulData from 10 dairy farms situated in different regions of Iran, where the majority of industrial dairy farms are situated, were utilized to calculate economic weights of traits. The production and reproduction records of cattle in all farms were tracked by ABCI. Performance of the cows was retrieved from a large collection of data given by ABCI that was collected by farmers through a survey, or it was calculated using cost and revenue modeling. Data sources used for deriving economic input parameters were based on the marketing circumstances in 2017 and 2018.Under Iranian dairy farm systems, the main dairy herd population comprises four major types of animals including calves, heifers, cows (both milking and dry), and fattened calves. Because it is related to feedlot and cattle production, the last group was excluded from the profit function. Profit, determined as total income minus total costs, was selected as the economic assessment criterion for the stated production system. Profit was expressed in terms of per cow calving per year as the reference unit. Iran’s currency is the Rial (Rl). Henceforth, the costs are stated in the US dollar, assuming an exchange rate of USD 1 = IRR 42,000. The following is the total profit function (Equation (2)):The impacts of genes expression may be observed as genetic development in diverse characteristics across time in various animal populations. Due to the potential cost of delayed gains, it is usually desired for genetic supremacy to be shown as soon as possible. This research updated the gene flow approach initially described by  for beefAccording to , the ecoThe first derivative of the profit function (Equation (1)) was selected considering milk, fat and protein yields for which the EV was required. Since there are no supply quotas for dairy farms in Iran, this is essentially equal to collecting the difference between the profits and costs of producing 1 kg of each product. Expenses associated with milk production were calculated by taking into account feed and labor, breeding, veterinary, fuel, housing and insurance costs. Only expenses related to metabolizable protein and net energy of lactation requirements were taken into account for protein and fat yields . The estThe EV for age at first calving was estimated according to . In addiThe EV of calving interval was calculated as the first partial derivative of the total profit function (Equation (1)), with respect to calving interval. Since cows express the trait just once throughout calving intervals and as a result have a relative gene expression coefficient of unity, the EW of calving interval is equivalent to the EV.In the present study, the EW was estimated to be USD −0.051 per kilogram of milk yield; USD 4.881 per kilogram of fat yield; USD 3.985 per kilogram of protein yield; USD −0.916 per day of age at first calving; and USD −1.695 per calving interval.The genetic-economic merit of 274,057 cows in Iran and its various climates was estimated using the LNI index as follows (Equation (3)):ijklmy and ijkmy are observation, μ is the overall mean, iH is the fixed effect of ith herd, jYS is the fixed effect of jth year-calving season, kR is the fixed effect of kth climate, lI is the fixed effect of lth selection index, (R × I)kl is the interaction between kth climate and lth selection index, kC is the fixed effect of kth sperm exporting country and ijklme and ijkme are random residual effects.The models used to analyze estimated breeding values (EBVs) and LNI index in Iran (Equation (4)) and each climate (Equation (5)) were as follows:The comparison of EBVs and LNI index of the sire groups was carried out by Tukey test using SAS 9.2 software . FinallyDescriptive statistics of investigated traits is presented in p < 0.01). Daughters of French sires had the highest EBVs for production traits  in Iran and semi-cold, moderate and warm climates. In the cold climate, the highest EBVs for milk and fat yields belonged to daughters of American sires, and for protein yield, the highest EBVs were estimated for daughters of German sires.The comparisons of EBVs for production traits between cows from different sire groups in Iran and cold, semi-cold, moderate and warm climates are shown in The Canadian selection indices have always had a high relative emphasis on the production traits ,8; howevp < 0.01). In Iran and semi-cold, moderate and warm climates, the daughters of French sires had the highest EBVs for the first calving age, and the daughters of German sires had the highest EBVs for the calving interval. Under cold climate, the daughters of German sires and French sires had the highest EBVs for the first calving age and calving interval, respectively. Since 2000, France included reproductive traits in selection indices of dairy cows in addition to the production traits. Thus, from 2001 to 2019, the relative emphasis on reproductive traits increased from 12.5% to 22%. Moreover, from 1997, selection indices in Germany have directly been focused on reproductive traits. However, the extent of this emphasis has always been relatively low so that the relative emphasis on these traits is only 10% today [Comparison of EBVs for reproduction traits between daughters from different sire groups in Iran and cold, semi-cold, moderate and warm climates is shown in 0% today . Applyin0% today .Comparison of least-square means of LNI index for the studied production and reproduction traits between cows from different sire groups in Iran and cold, semi-cold, moderate and warm climates is shown in As a result, it seems necessary to develop a specific genetic-economic selection index for each climate of Iran, and breeding and economic objectives of different regional climates should be considered.There were significant differences between the EBVs for milk, fat and protein yields, the first calving age and calving interval of sire groups (semen origins) in Iran and its various climates. In Iran, the highest and lowest EBVs belonged to French and German sires, respectively. In addition, the daughters of French sires in moderate climate had the highest EBVs regarding milk production, while the best performance of fat and protein production belonged to daughters of German sires in moderate and daughters of French sires in semi-cold climates, respectively. The lowest age at first calving was observed in calves of French sires in moderate climates, and lowest calving interval belonged to daughters of German sires in warm climates. Furthermore, in Iran and semi-cold, moderate and warm climates, the highest least-square means of the LNI index of progenies (genetic-economic merit) were estimated for French sires and in cold climate for German sires. As a result, it seems necessary to develop a specific genetic-economic selection index in each climate of Iran, and we recommended semen be imported based on breeding and economic objectives of different climates.


Three-dimensional printing technology enables the personalization and on-demand production of edible products of individual specifications. Four-dimensional printing technology expands the application scope of 3D printing technology, which controllably changes the quality attributes of 3D printing products over time. The concept of 5D/6D printing technology is also gradually developing in the food field. However, the functional value of food printing technology remains largely unrealized on a commercial scale due to limitations of printability and printing efficiency. This review focuses on recent developments in breaking through these barriers. The key factors and improvement methods ranging from ink properties and printer design required for successful printing of personalized foods  are identified and discussed. Novel evaluation methods for printability and printing precision are outlined. Furthermore, the design of printing equipment to increase printing efficiency is discussed along with some suggestions for cost-effective commercial printing. The existence of people with special food needs  has stimulated the development of innovative methods to design and create personalized foods . PersonaFood printing technology displays many advantages such as the ability to produce personalized food customization, digital nutrition matching, simplified supply chain, reuse of food scraps, and widening the source of food ingredients . These aIn terms of the modes of food printing, extrusion-based printing is the most widely used technique in the food sector . ExtrusiBased on extrusion-based printing technology, various personalized foods have been produced successfully to meet the unique needs of consumers. However, society requires innovative technology to not only maintain or enhance personalized value, but also to be sustainable and competitive. There are difficulties in scaling up personalized foods from the current level to industrial production. Printing accuracy and printing speed are major concerns for food printing technology. Several recent review articles published on food printing technology have focused on the impact of ink characteristics, printing parameters, and post-processing on printing formability at the household level ,10,11. TPrintability is defined as the similarity between the printed object and the printing model, and the sustainability of the printed object in the original structure and function during post-placement or post-processing . PrintabPrintability of printing inks is mainly reflected in rheological properties  during the printing process and shape stability during the deposition process ,14. PrinDysphagia refers to the impossibility of swallowing liquid, semisolid, or solid food. This condition affects almost 580 million people worldwide, especially infants and the elderly, and it leads to nutritional deficiencies . The dem2+ ions, which are then surrounded by galacturonic residues to form junction zones leading to a gel network [The combination of ions and hydrocolloids  is found to further improve printing accuracy and reduce the degree of collapse because ions enhance the network structure of hydrocolloids. Negatively charged carboxylic acid groups in hydrocolloids combine with positively charged Ca network .2+ to decrease the viscosity of the starch-pectin mixture while to improve its resistance to deformation, thus enhancing the printing performance. In real life, the printing method and the hydrocolloid species should be selected according to the degree of dysphagia.As shown in For protein-rich liquid foods like dairy products, in addition to adding hydrocolloids, a gel of protein itself under the action of enzymes is also a common way . Two typSpecially shaped foods enhance the interaction between diners and food materials. According to the path planned by slicing software, food printing technology successfully prints a variety of food shapes by layer-by-layer deposition. Although 3D printing technology can produce some shapes that are difficult to be replicated by molds, it is still difficult to print perfectly in shapes with few support points such as suspension, torsion, tilt, and hollow structures. Four-dimensional printing technology is an extension of 3D printing technology. In other words, 3D-printed products undergo controllable changes in sensory qualities  or nutritional qualities under external stimuli such as heat, pH, and light. Four-dimensional printing technology opens up ideas for the production of specially-shaped foods. Besides attracting consumers, specially shaped foods induced by 4D printing technology allow consumers to judge the cooking degree of foods according to sensory changes. It also fits well with the concept of “flat packaging”, which converts foods from 2D to 3D after processing, thus reducing packing costs and storage space . AccordiDehydration stimulation is also one of the common methods to induce the deformation of printing products. The difference in the dehydration rate is the main factor causing product deformation. The main factors causing variation in dehydration rate include the heating method and product formulation. Liu et al.  investigThe 5D printing concept is a recent technological innovation, which includes the movement of five axes, namely three axes of movement of the print head and two rotational axes of the print bed . CompareCompared to model-built products, 3D printing technology can influence changes in release rates while providing a higher degree of flexibility in dose size, shape, and appearance. Kamlow et al.  comparedBesides adding the released material directly to the printing ink used as a carrier, the released material can also be encapsulated in microcapsules and then added to the carrier. Compared with the method of direct addition to the hydrogel, microcapsules are more conducive to protecting functional substances. However, the development of microcapsules with a specific response to external stimuli is not perfect, and the migration rate of released substances in the carrier is slow, which lowers the product output. Currently, gelatin-Arabic gum composite coacervate is considered a potential wall material that can be used for 4D printing because of it is a thermo-responsive material . In addiIn conclusion, food printing technology provides a new idea for the controlled release of functional substances through the structural design of printing models in the porosity, the contact area with the stimulus source, and the spatial distribution of functional substances.The printing system mainly includes the mechanical part for implementing printing and the software part for controlling the printing path. The mechanical part mainly includes the extrusion device, transmission device, printing platform, and some auxiliary . Among tX, Y, and Z directions, and the movement modes of the printing head and printing platform can be summarized into three categories [X-axis and Y-axis directions, and the printing platform moves independently along the Z-axis direction. This form is suitable for printing a compact model; however, the normal discharge of the nozzle can be affected by the composite movement of the nozzle in the X-Y plane, thus reducing the printing quality. (2) The printing platform moves in the direction of the X-axis and the Y-axis, and the printing head moves independently in the direction of the Z-axis. This form requires precise control of Z-axis motion, high mass of the worktable, and lower printing speed. (3) The printing head/printing platform does composite motion in the direction of the Z-axis and X-axis (or Y-axis), which has small movement inertia and the overall structure is simple. This composite motion is widely used in the structural design of household 3D printing equipment. In order to include a variety of food materials in one printed object, the two-nozzle printer was developed [X, Y, and Z) of movement for the printing head used in 3D printing technologies, the five axes of 5D printing result from the two extra movements of the printing platform on two rotational axes [The Cartesian coordinate system is the traditional mode used in 3D/4D printing equipment due to its simple structure and easy manufacture. Common Cartesian coordinate systems involve movements in eveloped . Comparenal axes . High-prnal axes .The software part  is the command center controlling the mechanical part. Specifically, modeling software such as 123D, AutoCAD, and Rhinoceros are templates for printing . In ordeThe main role of slicing software is to split 3D models into 2D models and generate G codes based on the layering path, required speed, and amount of extrusion . Slic3r,The control system software can influence the accuracy of printed products through nozzle diameter, printing height of single-layer, printing speed, extrusion rate, filling rate, and filling pattern, alone or in combination . A nozzlSensory evaluation methodology is the most intuitive method to reflect printing accuracy. However, it cannot quantify the printing accuracy and cannot judge the internal situation of printed products . Shape fFormulation change can cause printability of printing inks to deviate from desired characteristics. Therefore, to save time, manpower, and material resources, it is important to have some means of evaluating the printability of food materials before printing. In addition, mathematical model-based simulation and equipment applied to food fields such as drying and detection are gradually introduced into the field of food printing to achieve a faster and more accurate evaluation of the printability of printing inks .The moisture content of printing inks can significantly affect the physical properties of food materials. The technical principle of low-field nuclear magnetic resonance (LF-NMR) was based on the spin-relaxation properties of hydrogen nuclei from water in a magnetic field . LF-NMR Mathematical modelling and simulation can be used to display the fluid characteristics of food inks in the extruder by solving the model equations . Guo et In addition to nutrition, color, taste, and texture as key elements of the consumption experience, the amount of food printing must be replicated to achieve commercial success. So far, the large-scale production of printed foods is still a future concept. Only increasing the speed and nozzle diameter of a printer will not only reduce the printing accuracy due to the presence of broken lines, line deformation, and expansion, but also fail to meet the requirements of commercial production. A potential approach involves using a multi-nozzle printer (number of nozzles ≥ 3) to make simultaneously multiple objects. The multi-nozzle printer referred to here has different designs according to single material printing and multiple material printing. For single material printing, the design of multiple-nozzle printers entails the connection of multiple single nozzles in series. Notably, the number of nozzles cannot be adjusted. After the design upgrade, the number of printing heads is modularized, and the number of printing heads can be added or reduced according to production needs. For multiple material printing, multiple groups of printing heads used for single material printing are added, and the added quantity of printing heads is consistent with the type of printed materials. The limitations of this idea are attributed to the following factors: lack of automatic feeding system, a limit on the number of printing heads, precise positioning of multiple printing heads, separation between printing and post-processing, automatic transmission system, slice quality of software, and quality monitoring, etc. .For improved slice quality of software, the researchers propose to improve the print speed by adaptive algorithms, which can adjust the print parameters to balance print quality and time . The proThree-dimensional food printing is a rapidly evolving technology, which makes it possible to produce innovative and unique products on demand. This technology is already well developed and applied commercially in many fields. For successful and cost-effective application in the food industry, much research and development are needed at the basic as well as the industrial scale to meet the special requirements unique to the food industry. The following recommendations are made on the basis of the relevant literature which is available.(1)Modification methods need to be researched in depth to improve printability of diverse printing inks and for smart food materials suitable for 4D/6D printing.(2)To minimize empirical trial and error tests, it is important to understand the fundamentals of the operation with appropriate numerical modeling. This is especially critical in view of the enormous variety of foods and their composition which affect their printing characteristics.(3)A food-grade coating with antibacterial properties has to be developed in order to prevent bacteria from accumulating in the corners of the printer and printers also need to use food-safety certified materials in order to prevent metal particles from migrating into printing inks.(4)Commercial utilization of 3D printing technology requires extensive testing for scale-up based on small-scale tests. Currently, printing technology can hardly compete with such food products that are produced by continuous processing lines, such as standard bakery products including buns rolls and bread. Automatic and intelligent printing equipment with multiple nozzles needs to be urgently developed.(5)Quality standards for food safety, color, aroma, texture, and composition of nutrients need to be established for personalized foods such as easy-to-swallowed foods, specially-shaped foods, and foods with controllable release of functional substances.Three-dimensional printing technology has great potential in manufacturing personalized foods. Four-dimensional printing technology adds a time dimension on the basis of 3D printing technology, which leads to controllable changes in the quality of printing products over time. In addition, 5D/6D printing technology is gradually introduced into the food industry because it can produce more complex shapes with less materials. However, printability and printing efficiency limit the large-scale development of food printing technology. Currently, the methods to improve printability and printing efficiency mainly include changing the physiochemical properties of printing inks, optimizing printing settings, and upgrading printer design. The method to improve the printing printability of different food materials needs to consider the personalized purpose of printing products. The integrated printing equipment including multiple nozzles, post-processing, and intelligent detection are ideal preferences to realize commercial printing. Additionally, algorithm simulation represented by mathematical models, and instrument measurement represented by rheometer, low field nuclear magnetic resonance technology, and infrared spectroscopy, etc., are new methods to quickly evaluate the printability of raw materials. The development and establishment of intelligent food materials; food safety coatings; efficient and intelligent printing equipment; and quality standards are needed to invest in more research in order to promote the commercial development of food printing technology.


The therapeutic alliance is critical to the efficacy of psychiatric treatment and can be weakened by involuntary treatment measures. In Western culture, mental illness is still associated with violence and if significant risk of violence is detected, in Spain a civil court can order the application of involuntary treatments such as Involuntary Outpatient Commitment.To discuss the effectiveness of some psychiatric involuntary treatments used in Spain.- Literature review about involuntary psychiatric treatments used in Spain - Case report about a patient undergoing Involuntary Outpatient CommitmentWe present the case of a 54-year-old man, diagnosed with schizophrenia, admitted to our acute psychiatric yard more than five times due to violent behavior and psychotic symptoms. Five years ago, he was summitted to a period of three years of Involuntary Outpatient Commitment. In Spain this measure can include the administration of involuntary medication, an injectable antipsychotic treatment in this case. At the end of the order, he immediately stops attending consultations and abandoned psychopharmacological treatment.Involuntary Outpatient Commitment is a controversial measure and it stirs up the concepts of stigma, coercion, care, patient autonomy and, globally, the values of humanization in psychiatry.No significant relationships.


Physiological aging triggers a cascade of negative effects on the human body and the human joint is only one of the several compartments affected by this irreversible and natural process. Osteoarthritis and cartilage degeneration can cause pain and disability; therefore, identifying the molecular processes underlying these phenomena and the biomarkers produced during physical activity is of critical importance. In the present review, the main goal was to identify and discuss the articular cartilage biomarkers analyzed in studies in which physical or sports activities were adopted and eventually to propose a standard operating procedure for the assessment. Articles collected from Pubmed, Web of Science, and Scopus were scrutinized to detect reliable cartilage biomarkers. The principal articular cartilage biomarkers detected in these studies were cartilage oligomeric matrix protein, matrix metalloproteinases, interleukins, and carboxy-terminal telopeptide. The articular cartilage biomarkers identified in this scoping review may aid in a better comprehension of where research on the topic is heading and offer a viable instrument for streamlining investigations on cartilage biomarker discovery. Aging is an irreversible and inevitable process that causes the progressive deterioration of several structures of the human body. One of the most affected tissues during aging is the cartilage; it is often the initial site of degenerative processes, a phenomenon most likely due to its reduced self-healing potential, particularly after repeated injuries . Aging iIn healthy joints, the cartilage has some ability to adapt to different types of mechanical stimulation , like dyIn the literature, there are numerous biomarkers used to evaluate the different aspects of cartilage/joint health, such as bone metabolism, molecules released within the synovium, and inflammation biomarkers ,26. HoweMany studies have been conducted over the years on OA biomarkers, identifying many options for evaluating its progression and severity; however, the literature has been widely heterogeneous when physical activity is an included variable. Considering the importance of regular physical activity for articular cartilage health, the broad heterogeneity of findings in published studies , and theThis review partially followed the preferred reporting for systematic reviews and meta-analyses for scoping reviews (PRISMA-ScR) checklist and explanation . The proThe search was performed on Pubmed, Web of Science, and Scopus until 21 September 2022, and all articles published were considered regardless of the date of publication. Keywords for the subject , intervention , and outcomes  were connected with the Boolean operators AND and OR. The following string was adopted in the three databases searched:(“articular cartilage” OR fibrocartilage OR gristle) AND  AND (collagens OR chondron OR matrix OR collagen OR proteoglycan OR synoviocyte).The population investigated had to be composed of active or sportive individuals, and the intervention and the terms of comparison adopted had no restrictions. The outcomes considered were only biomarkers classified as “burden of disease” . RegardiTwo investigators performed the collection and screening against the eligibility criteria. The screening was performed by title, abstract, and full text. Manuscripts were retrieved also by checking the references of the included papers.All the collected papers underwent narrative analyses, and relevant biomarkers were then presented and discussed.A total of 4342  articles were found in the three databases searched. After duplicate removal, a total of 2685 articles were screened against the eligibility criteria. Following the exclusion of papers about animals or direct interventions on the cartilage , about treatments other than physical or sports activities , or in injured people , a final number of 24 studies were included. The flow diagram can be found in the The main burden of disease biomarkers identified in the scoping review were the cartilage oligomeric matrix protein (COMP), detected in 16 studies, the carboxy-terminal telopeptide (CTX-II) in 5 studies, interleukins (ILs) in three studies, the matrix metalloproteinases (MMPs) in 2 studies. Several other biomarkers have been analyzed in different studies: in one study, circulating levels of serum C-reactive protein (CRP) combined with cartilage type II collagen measurement was used as a parameter of general inflammatory reaction and cartilage metabolism in marathon runners . SimilarA total of 16 studies investigated the sCOMP. Most of the studies investigated the articular biomarker after a running intervention (seven times), followed by walking (four times), and general (three times) exercises. Drop landing and soccer interventions were studied two times, while resistance exercises, jumping exercises, cycling, volleyball, basketball, weightlifting, and swimming one time.COMP is a 535-kDa non-collagen extracellular matrix component of the thrombospondin family proteins and it is primarily found in the articular cartilage, tendons, and synovium . It is aThe level of sCOMP increases immediately after physical activity and transiently elevated levels have been detected in athletes from different disciplines such as running ,45,46,47It was found that in people with OA, a combination of aerobic, muscle strengthening, and flexibility exercises can increase sCOMP concentrations . FurtherMMPs were studied in only two studies, one on ultramarathons  and one A total of five studies adopted CTX-II: general exercise and swimming were proposed two times while resistance training, running, cycling, and futsal only one time. CTX is a fragment of the telopeptide of collagen, and they both derive from the bone, type I (CTX-I), and the cartilage, type II (CTX-II). While CTX-I is a serum biomarker of bone turnover, CTX-II is a biomarker of articular cartilage degradation . It is oILs were proposed in five studies, endurance activities in three studies, while two studies proposed an exercise routine. Interleukins are well known inflammatory markers. IL-10 is a cytokine with chondroprotective anti-inflammatory and anti-apoptotic properties . ConsideDifferent molecular burdens of disease biomarkers have been investigated to address the effects (positive or negative) of physical activity on the articular cartilage and the most commonly adopted ones are COMP, certain MMPs, CTX-II, and a number of ILs. All of them display different trends in response to exertion and may be useful for determining various aspects of cartilage health. Amongst these, the most frequently adopted is COMP. Considering the findings, a standard operating procedure could consist of taking sCOMP samples always at the same time of day. It is important to specify the hours and the time elapsed since the last training. It is fundamental to describe the participants’ characteristics, not only in terms of disease but also their diet and working experience. A final aspect to consider is the need to deeply describe the activity practiced in terms of frequency, time, type, and volume. It is also important to collect data about their physical background. Only if all this information is provided, will it be possible to compare the studies in the future and improve the quality of the research in this field. A synthesis of the proposed standard operating procedure is presented in Data from several recent reports ,85,86 poPhysical activity is fundamental to maintaining cartilage health and homeostasis . Indeed,This study aims to provide an overview of the principal biomarkers adopted in the evaluation of the articular cartilage in healthy people and in patients with OA. The ultimate goal is to provide researchers but also experts in the sector with an instrument to better understand and standardize the use of these biomarkers in future studies that evaluate cartilage and the effects of physical activity. In this way, it will be possible to compare the data and to try to create guidelines to establish the correct frequency, intensity, duration, and type of activity for achieving the best outcomes for cartilage health in the healthy and diseased populations. This review could represent a valid tool for researchers to acknowledge the current state of the field of cartilage biomarkers mainly employed in studies on cartilage and physical activity.There is a huge variety of molecular biomarkers currently under investigation to reveal the conditions of articular cartilages, either after simple mechanical stimulation and/or during/after physical activity. The most frequently adopted markers are sCOMP, CTX-II, Ils, and MMPs, and their use in combination seems to provide sufficient data to determine whether crucial protective and regenerative mechanisms are activated within weight-bearing joints by physical exertion both in healthy people and in patients with OA. A standard operating procedure for a burden of disease molecular biomarker evaluation of the cartilage during physical activity practice has been proposed.


Whereas significant strides have been made in the treatment of lysosomal storage diseases (LSDs), the neuronopathy associated with these diseases remains impervious mainly because of the blood-brain barrier (BBB), which prevents delivery of large molecules to the brain. However, 100 years of research on the BBB since its conceptualization have clarified many of its functional and structural characteristics, spurring recent endeavors to deliver therapeutics across it to treat central nervous system (CNS) disorders, including neuronopathic LSDs. Along with the BBB, the cerebrospinal fluid (CSF) also functions to protect the microenvironment of the CNS, and it is therefore deeply involved in CNS disorders at large. Recent research aimed at developing therapeutics for neuronopathic LSDs has uncovered a number of critical roles played by the CSF that require further clarification. This review summarizes the most up-to-date understanding of the BBB and the CSF acquired during the development of therapeutics for neuronopathic LSDs, and highlights some of the associated challenges that require further research. Lysosomal storage diseases (LSDs) are progressive inherited disorders of the metabolism, in which, due to a genetic deficiency of lysosomal enzymes, substrates remain unmetabolized and accumulate inside the organelles of the endosomal-autophagic-lysosomal system. About two-thirds of patients with LSDs exhibit complex central nervous system (CNS) dysfunctions. Such neuronopathic LSDs include mucopolysaccharidoses , sphingolipidoses, mucolipidoses, oligosaccharidoses, multiple sulfatase deficiency, and neuronal ceroid lipofuscinoses Sun .The recent advent of novel treatments for LSDs  has benefited patients by alleviating their symptoms and improving prognoses  both play critical roles in preserving the intricate structures and functions of the brain by protecting it from external insults, and from pathogenic and toxic substances Proulx . UnfortuHowever, better understanding and therapeutic utilization of the BBB and the CSF are of paramount importance for the development of novel therapeutics to address CNS disorders in patients with neuronopathic LSDs. A wide range of issues need to be addressed, from clarifying the underlying pathogenesis of neurodegeneration, developing and manufacturing BBB-crossing enzymes, to evaluating the optimal drug effects against CNS symptoms in clinical trials. Recent research into the BBB and CSF has been delineating the complex physiological nature of these fundamental mechanisms, which are still far from being fully elucidated. This review attempts to summarize recent advances in our understanding of the BBB and the CSF which need to be taken into account during research aimed at developing effective treatments for neuronopathic LSDs.General structures and functionsOriginally conceptualized by Lewandowsky  followinDysfunction of the BBB is known to play a central role in a number of neurodegenerative diseases (e.g. Alzheimer’s disease and multiple sclerosis), whereby the disrupted BBB causes ion dysregulation and allows neurotoxic blood-derived debris, cells, and microbial pathogens to enter the brain; this leads to inflammation, opening up multiple possible pathways towards neurodegeneration Transcytosis through the BBB – intracellular transport in cerebrovascular endothelial cells –Table Perhaps due to the specialized tight junctions that prevent paracellular passage between the blood and the brain, transcellular pathways have been a major focus of attention as a means of drug delivery. In particular, transcytosis  is considered critical in overcoming the restrictive properties of the BBB Drug delivery to the brain via receptor-mediated transcytosisTranscytosis is initiated by endocytosis, a process by which substances in the luminal spaces are internalized by endothelial cells, and the internalization process can proceed via three pathways: the caveolae, clathrin-dependent endocytosis, or clathrin-independent endocytosis  for neuronopathic LSDs. Five compounds that harness receptor-medicated transcytosis have so far progressed to clinical trials Table .Table 2CAs an example, a schematic illustration showing the mechanism of transcytosis via transferrin receptors is given in Fig. Transferrin binds to the transferrin receptors located on the luminal side of the microvascular endothelial cells in the brain and is absorbed into the endothelial cells (endocytosis), in which it is then transported towards the abluminal side of the cell facing the brain parenchyma, and subsequently released from the receptors to reach the brain parenchyma (exocytosis). Likewise, enzymes fused with an anti-transferrin receptor antibody bind to the transferrin receptors, are absorbed into the endothelial cells, and are then released into the abluminal side of the cells so that they can diffuse into the brain parenchyma.There have been a number of translational challenges in establishing drug delivery via receptor-mediated transcytosis. One of them is that using receptor-mediated transcytosis of targeted enzymes can affect the original innate transcytosis that is supposed to deliver physiological substances like transferrin , α-L-iduronidase fused with anti-insulin receptor antibody, in patients with MPS-I detected drug-related transient hypoglycaemia in 6.4% of the patients , iduronate-2-sulfatase fused with anti-transferrin receptor antibody, on patients with MPS-II, there were no adverse events associated with the iron metabolism, which it was thought the drug might affect -containing glycans on the lysosomal enzyme that mediate enzyme uptake by the M6P receptor are present on most cells in the body; hence the resultant somatic efficacy of the fusion protein. The affinity of the M6P-containing glycans to the M6P receptor is in the low nM range, ensuring rapid cellular uptake of the enzyme  active CSF formation (secretion), 2) passive CSF absorption (drainage), and 3) unidirectional CSF flow from its formation sites to absorption sites  that is almost antithetical to the traditional view has recently been put forth  that remain uncatabolized because of the enzyme deficiency seen in patients with neuronopathic LSDs are by far the most important determinant in initiating the complex cascade of neurodegenerative processes (Sato and Okuyama In an established animal model of MPS II, close correlations were observed between intracerebral and CSF HS levels, and intravenous administration of pabinafusp alfa led to dose-dependent reductions in HS levels in the CSF, along with corresponding reductions in HS levels in the brain (Sonoda et al. IT administration of enzymes in patients with MPS-II (Muenzer et al. The blue arrows represent the innate natural circulation of the CSF, the green arrows intravenously administered BBB-crossing enzymes, the black arrows intrathecally administered conventional enzymes, and the yellow ones intracerebroventricularly administered conventional enzymes (Modified from Tumani et al. IT administration of enzymes can, by definition, directly affect the substrate levels in the subarachnoid space, while their penetration into the brain parenchyma to act against CNS disorders is very limited, as discussed above. In contrast, BBB-crossing enzymes administered intravenously can traverse the BBB via receptor-mediated transcytosis, thus addressing the brain parenchyma widely and directly, where they reduce the substrate accumulations therein. The enzymes are thought to be mostly consumed in the process, so that few, if any, remnants circulate into the CSF. Therefore, changes in CSF substrate levels brought about by intravenous ERT with BBB-crossing therapeutics truly reflect the changes in intracerebral substrate levels.sui generis of the CSF and BBB have to be taken into account when evaluating the pharmacokinetics and pharmacodynamics of drugs targeting the CNS.As shown in Fig. The year 2021 marks the centenary of the naming of the BBB by Lina Stern (1878–1951; Dreifuss and Tikhonov However, there are still a number of remaining issues surrounding the emerging ERTs for neuronopathic LSDs. First, details of the specific regulating processes in various modes of transcytoses need to be clarified so that drug delivery via transcytosis can be optimized to consolidate efficiency, efficacy, and safety. Second, the CSF microcirculation has only been partly elucidated, and its pharmacokinetic and pharmacodynamic relevance to ERT and other treatment modalities for neuronopathic LSDs needs further investigation. Third, knowledge about how the neuronopathy seen in patients with LSDs responds to treatment is still scant, which means that valid and reliable methods of evaluating the efficacy of novel therapeutics are yet to be established. In other words, with the advent of new therapies to address the neurological and psychiatric signs and symptoms of patients with neuronopathic LSDs, we need to learn to understand how these new treatments influence an affected individual in the way he/she feels, functions, and survives. Also, the inevitable paucity of patient populations and significant heterogeneity in the clinical manifestations and prognoses of these rare diseases impede the efficient conduct of clinical trials and cause further delays in introducing novel treatments. Last but not least, designing and manufacturing a fusion protein for efficient transcytosis-mediated drug delivery entails formidable challenges in establishing acceptable pharmacokinetic and pharmacodynamic profiles, stability, yield, and other production-related technical specifications.It is hoped that the wide-ranging research being carried out will lead to further insights into neuronopathic LSDs, facilitating the development of effective treatments for patients suffering from these rare, debilitating diseases.


Inflammatory bowel disease (IBD) is a gastrointestinal disorder, affecting about several million people worldwide. Current treatments fail to adequately control some clinical symptoms in IBD patients, which can adversely impact the patient’s quality of life. Hence, the development of new treatments for IBD is needed. Due to their unique properties such as biocompatibility and sustained release of a drug, biomaterials-based drug delivery systems can be regarded as promising candidates for IBD treatment. It is noteworthy that considering the pathophysiological changes occurred in the gastrointestinal tract of IBD patients, especially changes in pH, surface charge, the concentration of reactive oxygen species, and the expression of some biomolecules at the inflamed colon, can help in the rational design of biomaterials-based drug delivery systems for efficient management of IBD. Here, we discuss about targeting these pathophysiological changes using biomaterials-based drug delivery systems, which can provide important clues to establish a strategic roadmap for future studies. IBD is a group of idiopathic, chronic inflammatory disorders of the gastrointestinal tract which includes two major forms, ulcerative colitis (UC) and Crohn’s disease (CD) . It is rMultiple environmental agents, involved in the development of IBD, include factors like infectious pathogens, diet, and autoantigens residing in the gut microbiome . It is avia biomaterials-based drug delivery systems  that can provide key clues to establish a strategic roadmap for future studies.At present, existing therapies can’t adequately control clinical symptoms in a significant number of IBD patients. Consequently, novel treatments for IBD is needed . It is wvia the gastrointestinal tract, and hepatic first-pass metabolism following the intestinal absorption process. The most important benefit of rectal administration  is topical drug delivery to the injured colon. However, maintaining the enema or suppository for long periods of time in colon is the main limitation of rectal administration. Intravenously administered drugs do not undergo hepatic first-pass metabolism, which it is the main advantage of intravenous use. Despite this benefit, intravenous administration has some challenges like nonspecific distribution in normal tissues and short blood circulation time , corticosteroids, immunomodulators (such as azathioprine and methotrexate), Tofacitinib (a Janus kinase inhibitor), TNF-α blocking biological agents , and anti-integrin biological factors  . For theion time . Hence, ion time .The most important pathophysiological changes in the gastrointestinal tract of the patients with IBD include changes in pH, surface charge, the concentration of reactive oxygen species (ROS), and the expression of some biomolecules at the inflamed colon. In normal physiological conditions, colonic luminal pH is in the range of 7-8, which is affected by many factors especially bicarbonate and lactate production . ColonicAnother important pathophysiological factor in the colon of IBD patients is surface charge. In normal physiological conditions, surface charge of colon epithelium is negative . MucosalThe expression of some biomolecules like inflammatory biomarkers in the injured colon is increased during IBD development compared with their basic expression level in normal physiological conditions . The oveConsidering the pathophysiological changes which occurred in the gastrointestinal tract due to IBD can help in the rational design of biomaterials-based DDSs for efficient management of IBD. In particular, changes in the inflamed regions of the colon can be a good clue to specifically targeting the inflamed area rather than the whole colon. A summary of studies on targeting pathophysiological changes using biomaterials-based DDSs is presented in in vitro release study. The oral administration of this nanocomplex showed great accumulation of the nanocomplex as well as the release of infliximab in the injured colon of DSS-induced colitis mice. In addition, pro-inflammatory factors (including IL-6 and TNF-α) were decreased after the oral use of the nanocomplex  score after the oral use of the nanoparticles in mice with colitis induced by DSS, indicating the good functionality of the nanoparticles in the alleviation of colitis symptoms , liposomes as one of biomaterials-based DDSs are promising candidates for the encapsulation of diverse drugs including both hydrophobic and hydrophilic compounds. Interestingly, negatively charged liposomes showed preferential adhesion to the inflamed colon of dinitrobenzenesulfonic acid-induced colitis rats via electrostatic interaction. They also had a two-fold higher cumulation in the inflamed regions compared with cationic and neutral liposomes, therfore negatively charged liposomes can be valuable to deliver anti-inflammatory drugs to the inflamed colon  in the colon’s inflamed regions, indicating the good functionality of this hydrogel for local targeting the damaged areas of the colon. Moreover, this enema exhibited the ability of accelerating mucosal healing via boosting Syndecan-1 (Sdc-1), a tissue repair agent, and can also help in the prophylaxis of inflamed colon infection via physical barrier formation -b-(N-acryloylthiomorpholine)] through polymerization induced self-assembly. These micelles contain thioether groups in the hydrophobic core, which are susceptible to oxidation in the presence of ROS. The oxidation of thioether groups leads to the formation of hydrophilic sulfoxide groups and the degradation of micelles. Furthermore, the micelles were loaded with fluorescent dye Nile red to monitor the micelles degradation. When the micelles were incubated with monocytes isolated from patients with IBD, which had increased ROS production, the degradation of micelles was successfully occurred in comparison with the micelles incubated with monocytes isolated from healthy individuals  score and an increase in the colon length in mice with colitis induced by DSS, showing the great functionality of the micelles in the alleviation of colitis symptoms  phenylboronic acid pinacol ester and tempol moieties are valuable candidates for the conjugation to biomaterials for the preparation of ROS-responsive biomaterials-based DDSs . In one via biomaterials-based DDSs functionalized with ROS-sensitive groups can be a good potential candidate to deliver potent drugs to the inflamed colon. However, the following points should be considered before the clinical application of such ROS-sensitive systems. Until now, just the cytotoxicity phase of these ROS-responsive systems was investigated. Therefore, a comprehensive evaluation of the safety of these systems should be performed in several animal models because biocompatibility is an essential factor in the entry of new therapeutic systems into the clinical trial phase. Moreover, in normal concentrations of ROS, these systems should be inactive  and pro-inflammatory  agents  agents . In a si) agents . Further) agents . From a ) agents .via biomaterials-based DDSs functionalized with specific ligands of these biomolecules can be a promising option for delivery of drugs to the injured colon (x (a tetrasaccharide) is one of specific ligands of P-Selectin , which can help bowel regeneration and has a protective role against IBD. The uptake of these particles was successfully occurred by the inflamed human umbilical vein endothelial cells. Furthermore, the intravenous injection of these liposomes showed high accumulation of them in the inflamed colon of DSS-treated mice, leading to reduction of inflammation  is considered one of the most important causes of healthcare-associated infections (Clostridium difficile to inflamed colon epithelium, it can be a prophylaxis strategy for inflamed colon infection. Hydrogel-based enema as one of the most attractive biomaterials-based DDSs for local drug delivery can fulfill this aim because it adheres to the inflamed colon epithelium and forms a physical barrier (Furthormore, it is important to point out that the patients with IBD are susceptible to infection, especially fections . If ther barrier . Hence, The most important pathophysiological changes at the inflamed colon include changes in pH, surface charge, the concentration of reactive oxygen species, and the expression of some biomolecules. Targeting pathophysiological changes in IBD patients using biomaterials-based DDSs is a promising option for the effective delivery of drugs to the injured colon. However, IBD is a multifactorial disease, so developing biomaterials-based DDSs through single targets may not be effective in the treatment of IBD. For this reason, the development of versatile biomaterials-based DDSs with ability in the simultaneous targeting of multiple agents of IBD may be regarded as a valuable and potential candidate for more efficient management of IBD in future studies.