Diabetic ulcer is a serious complication of diabetes. Compared with that of healthy people, the skin of patients with a diabetic ulcer is more easily damaged and difficult to heal. Without early intervention, the disease will become increasingly serious, often leading to amputation or even death. Most current treatment methods cannot achieve a good wound healing effect. Numerous studies have shown that a nanocomposite hydrogel serves as an ideal drug delivery method to promote the healing of a diabetic ulcer because of its better drug loading capacity and stability. Nanocomposite hydrogels can be loaded with one or more drugs for application to chronic ulcer wounds to promote rapid wound healing. Therefore, this paper reviews the latest progress of delivery systems based on nanocomposite hydrogels in promoting diabetic ulcer healing. Through a review of the recent literature, we put forward the shortcomings and improvement strategies of nanocomposite hydrogels in the treatment of diabetic ulcers. Diabetes mellitus is a chronic disease caused by the disorder of glucose metabolism. Long-term hyperglycaemia will increase the blood viscosity of patients, weaken the elasticity of blood vessels and weaken the local skin barrier function of patients, resulting in diabetic ulcers such as diabetic foot ulcers, leg ulcers, and buttock pressure ulcers . For norAt present, there are a variety of clinical methods to treat diabetic ulcers, mainly based on blood glucose control, timely dressing, debridement and prevention and control of infection . HoweverWith the continuous development of biomedical materials and in-depth studies of wound healing theory, investigators found that a clean and humid environment is more conducive to wound healing. Therefore, wet wound dressings, particularly hydrogels have attracted attention . A hydroTo overcome these limitations, researchers worldwide compete to conduct studies aimed to improve the mechanical properties of hydrogels. With the continuous development of nanotechnology, researchers have found that nanocomposite hydrogels can be prepared by introducing nanostructured particles into the hydrogel network through chemical bonding or physical adsorption. These methods can improve the mechanical strength of hydrogels and impart some new properties to hydrogels . SpecifiIn particular, nanocomposite hydrogels are advantageous, because they can be loaded with small-molecule drugs, biomacromolecules, inorganic nanoparticles and other active substances that enhance the biological activity of nanocomposite hydrogels and further expand their application to wound management . When a Nanohydrogels have attracted increasing attention because of their great potential for use as effective drug delivery systems. This review focuses on the promotion of ulcer healing in animal models of diabetes or patients with diabetes through loading small-molecule drugs, protein drugs, silver nanoparticles and other substances. Furthermore, we evaluated the challenges to translating these nanocomposite hydrogels to the clinic.There are many advantages in using nanocomposite hydrogel as a carrier of small-molecule drugs: 1) It can protect drugs and improve their stability . 2) It cin vitro. Further, the hydrogel upregulates vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α, which can accelerate the healing of a diabetic ulcer (Deferoxamine (DFO), a small-molecule drug approved by the Food and Drug Administration (FDA) of the United States, has been used to induce angiogenesis in bone and skin regeneration . DFO canic ulcer .4-Hydroxy-3-methoxycinnamic acid (ferulic acid) is a natural antioxidant with antibacterial activity. It can reduce blood glucose levels, scavenge free radicals, promote angiogenesis and so on . It was 3-Methyl-1-phenyl-2-pyrazolin-5-one (edaravone) strongly scavenges free radicals and promotes wound healing, although its poor stability and solubility hinders its local application . It was Berberine has anti-inflammatory, antibacterial and hypoglycaemic effects, promoting the healing of diabetic ulcers . However7-Glucuronic acid 5,6-dihydroxyflavone  can promote the expression of epidermal growth factor and VEGF and then enhance the regenerative ability of epidermal cells, endothelial cells, and fibrotic tissue, to induce wound healing . It has Curcumin has been shown to promote wound healing . HoweverPrevious studies have confirmed that quercetin or oleic acid alone can promote wound healing . A recenAt present, most studies focus on the use of protein hormones and growth factors to promote the healing of diabetic ulcers . CompareProteins are important nutrients for the proliferation and remodelling of wound healing . ProteinInsulin is a physiological hypoglycaemic agent, which is commonly used in the treatment of diabetes. Recent studies have found that local use of insulin can treat diabetic ulcer . Both frin vitro and the secretion of VEGF. Protamine nanoparticles and hyaluronic acid oligosaccharides synergise to promote wound healing  is a low-molecular-weight peptide composed of 53 amino acids . Previouiguanide . The hydiguanide . It can Becaplermin containing a human recombinant platelet-derived growth factor BB (PDGF-BB) gel for the treatment of diabetic foot ulcers. A recent study reported that PDGF-BB encapsulated with a self-assembled peptide (RADA 16-I) can create a kind of nanohydrogel with good biocompatibility and biodegradability, which allows continuous delivery of PDGF-BB and destroys the bacterial biofilm , known as “trauma factor”, is mainly stored in platelet alpha granules . When th biofilm . CompareStromal cell-derived factor-1α (SDF-1α) is a key chemokine involved in the regulation of tissue and organ injury and repair with strong chemotaxis on diverse inflammatory and mesenchymal stem cells . SDF-1α Moreover, combining small-molecule drugs with growth factors may be a promising method to promote wound healing. The curcumin and EGF modified by nanotechnology were encapsulated in hydrogels, which could release curcumin and EGF on demand and synchronise with wound healing. Specifically, curcumin is rapidly and continuously released during the early stage of wound healing to alleviate inflammation and oxidative stress, while EGF is relatively slow-acting and sustains late proliferation and extracellular matrix remodelling . A similBacterial infection is an important factor that hinders wound healing of diabetic ulcers . The decMost metal nanoparticles possess antibacterial effects, among which that of silver nanoparticles is the strongest. Silver nanoparticles have excellent antibacterial properties through controlling the slow release of silver ions from their surface . In addiin vitro. The chitosan polyethylene glycol hydrogel impregnated with silver nanoparticles can slowly and continuously releases silver nanoparticles within 7 days, can accelerate the re-epithelisation and collagen deposition of diabetic wounds of rabbits and have better wound healing ability (Recent studies have shown that bamboo cellulose nanocrystals can be impregnated with silver nanoparticles to obtain inexpensive nanocomposite hydrogels . The lev ability .+ and CD3+ were upregulated (+ and eliminate adsorbed bacteria by interacting with sulfur-containing proteins on bacterial membrane. Furthermore, Recently, it has been reported that a novel antifouling and anti-infective hydrogel can be prepared by mixing silver nanoparticles with chitosan and dextran. After treatment with this hydrogel, the wounds of diabetic SD rats contracted rapidly, and the expression levels of CD68egulated . It is s+ will have adverse side effects on human cells and tissues (2S quantum dots was reported (However, the latest research shows that silver nanoparticles readily agglomerate, and high concentration of Ag tissues . Based oreported . It can Exosomes are lipid bilayer extracellular vesicles secreted by various cells, which contain a series of bioactive substances such as proteins, lipids, RNA and DNA. They can promote angiogenesis, stimulate collagen deposition, inhibit inflammation and accelerate wound healing . PreviouIn vivo studies showed that the hydrogel significantly promotes the healing of full-thickness skin wounds by enhancing granulation tissue formation, promoting collagen deposition and accelerating re-epithelialisation and neovascularisation.Recent studies have shown that platelet-rich plasma comprising numerous cell growth factors can accelerate wound healing by regulating cell proliferation . HoweverOxygen is the most important nutrient for cell survival. Insufficient oxygen delivery prevents cell migration and angiogenesis and reduces cell growth and differentiation, thereby delaying wound healing . RecentlIncreasing evidence shows that mesenchymal stem cells can promote wound healing of diabetic ulcers by promoting cell proliferation and differentiation and extracellular matrix synthesis, releasing growth factors and promoting angiogenesis . RecentlRecently, researchers have found that proteins in egg white play an important role in wound healing . Based oDiabetic skin ulcers are a serious complication caused by the internal environmental changes and the local pathological changes of skin caused by diabetes. Many studies have shown that a nanocomposite hydrogel is an ideal local drug delivery method to promote the healing of diabetic ulcers and has achieved satisfactory therapeutic effects. Nanocomposite hydrogels have the advantages of small particle size, high water content, long drug released property and good biocompatibility and biodegradability. They can also be used for the delivery of small-molecule drugs, protein drugs and silver nanoparticles to promote the healing of diabetic ulcers. In conclusion, nanocomposite hydrogels are expected to serve as a new dressing for large-scale clinical treatment of diabetic ulcer patients.However, there are still problems to be solved. For example, most nanocomposite hydrogels are in the early stage of experimental or clinical development, and there is a lack of large-scale clinical studies to establish efficacy and safety. The preparation of some hydrogels is complex, which makes it difficult to widely apply them to the clinic. To reduce the adverse reactions in clinical treatment, it is necessary to accurately control a series of properties of nanocomposite hydrogels, such as crosslinking degree, porosity, swelling, mechanical properties, cell adhesion and permeability and to simulate the extracellular matrix microenvironment as much as possible to maintain the characteristics and activity of each component. Furthermore, it is important to determine the optimal moisture removal rate of the composite hydrogel and to determine the optimal moisture content of the composite hydrogel for patients with diabetes. It can be predicted that our research on nanocomposite hydrogel materials will be more detailed, and the preparation methods will be more advanced. More new nanocomposite hydrogel carriers will be developed and applied to clinical drug delivery, which will benefit more patients with chronic ulcers.


Aging-associated conditions, such as cardiovascular (CVD) and respiratory diseases, represent significant public health concern , 2. ImpaSinha et al., primary analyses from 17,049 subjects revealed a significantly higher risk of incident coronary heart disease in patients with systemic lupus erythematosus (SLE)  and systemic sclerosis (SSc)  as compared to subjects without chronic inflammatory conditions. Surprisingly, patients with HIV, psoriasis (PSO), rheumatoid arthritis (RA) and inflammatory bowel disease did not show the same increased risk. The observed discrepancies with previously published studies . In the era of SARS-CoV-2 pandemic exploring multisystemic biologic interactions dependent on gender is critical for effective treatment and disease prognosis.In the review paper by Rastogi et al. elaborated on maternal obesity, which is linked to long-term morbidities, such as cardiac and pulmonary conditions, in the offspring. Besides known immune and metabolic effects associated with obesity, the authors highlighted a role of oxidative stress and microbiome disbalance during pregnancy and what future preventive and treatment measures should be applied to both mothers and children in order to prevent the cardio-pulmonary continuum from development throughout the life course.By further investigating the above mentioned gender differences, Lorey et al.As we tried to emphasize with the current collection of research articles, one of the reasons underlying the reported pathophysiological association between cardiovascular and pulmonary diseases resides in persistent inflammation and impaired immune response. Therefore, this impaired biological multisystem interaction also responsible for the lipoprotein metabolic dysregulation observed in chronic inflammatory conditions like PSO  and COPDFilip et al. in a model of ApoE−/−AOC3−/− double knockout mice and postmortem human coronary plaque samples. The authors discovered that absence of amine oxidase copper containing 3 (AOC3) enzyme, involved in vascular smooth muscle cell phenotype switching, had a pro-atherosclerotic effect and led to atheroma progression.Finally, new insights into atherosclerosis biology and plaque formation have been elegantly demonstrated by In summary, this collection of research articles aimed to raise awareness among researchers and health care specialists of the complex interplay among pulmonary and cardiovascular diseases with immune-inflammatory component.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.


The diet and gut microbiota have been extensively interrogated as a fuel for gut inflammation in inflammatory bowel diseases (IBDs) in the last few years. Here, we review how specific nutrients, typically enriched in a Western diet, instigate or deteriorate experimental gut inflammation in a genetically susceptible host and we discuss microbiota-dependent and independent mechanisms. We depict the study landscape of nutritional trials in paediatric and adult IBD and delineate common grounds for dietary advice. Conclusively, the diet reflects a critical rheostat of microbial dysbiosis and gut inflammation in IBD. Dietary restriction by exclusive enteral nutrition, with or without a specific exclusion diet, is effectively treating paediatric Crohn’s disease, while adult IBD trials are less conclusive. Insights into molecular mechanisms of nutritional therapy will change the perception of IBD and will allow us to enter the era of precision nutrition. To achieve this, we discuss the need for carefully designed nutritional trials with scientific rigour comparable to medical trials, which also requires action from stake holders. Establishing evidence-based dietary therapy for IBD does not only hold promise to avoid long-term immunosuppression, but to provide a widely accessible therapy at low cost. Identification of dietary culprits disturbing gut health also bears the potential to prevent IBD and allows informed decision making in food politics. The diet and host immune responses determine gut microbial composition and function.Excessive intake of specific macronutrients enriched in a Western diet promotes experimental gut inflammation by perturbation of host–microbe commensalism.Dysbiosis in inflammatory bowel diseases (IBDs) is fuelling experimental gut inflammation.Clinical trials indicate that the diet affects gut inflammation in patients with IBD.This review summarises recent experimental and clinical advances on the role of the diet in IBDPatient-tailored dietary advice will be a cornerstone to prevent and treat IBD in the future.Inflammatory bowel diseases (IBDs) encompass a spectrum of chronic inflammatory disorders in and beyond the gut, typically referred to as Crohn’s disease (CD) or ulcerative colitis (UC).−/−IL10), by the bloom of the gut pathobiont Bilophila wadsworthia.−/−IL10 mice and toxic colitis,−/−IL10 mice.−/−IL10 mice.Western dietary habits are characterised by increased intake of fat and simple carbohydrates, and reduced intake of plant-derived complex carbohydrates . Recent experimental evidence indicated that specific macronutrients in a Western diet deteriorate experimental gut inflammation that is induced by genetic or chemical means . Prime eBox 1A Western diet is enriched with simple carbohydrates  and fat , while being largely devoid of fibre.Western dietary constituents also directly impact gut mucosal immune responses. For example, we recently demonstrated that long-chain polyunsaturated fatty acids (PUFAs), contained in red and white meat, eggs and cooking oils, trigger an inflammatory response from gut epithelial cells, which is restricted by Glutathione peroxidase 4 (GPX4).Collectively, these studies demonstrated that excessive intake of specific nutrients and additives in a Western diet, such as PUFAs, simple carbohydrates and food colourants, trigger or deteriorate experimental gut inflammation, by exploiting the gut microbiota, or by engaging innate immune receptors and related cellular stress signalling . As suchFirmicutes and increased abundance of Proteobacteria and Bacteroidetes phylum members in stool, with specific species and related metabolic pathways emerging.In IBD, altered microbial signatures  have been consistently reportedThe diet is thought to strongly contribute to microbial variation in mice (~50%) and humans ~20%, with strong differences between individuals.Butyrococcus pullicaecorum, improved epithelial barrier integrity in CD based on simulations.Carbohydrates are generally classified as digestible and non-digestible and are contained in a wide range of food items. Digestible carbohydrates can be enzymatically degraded into simple sugars that are largely absorbed in the small intestine and passed into the bloodstream through the portal vein.Bacteroides.Human studies indicated that a high-fat diet increases anaerobic abundance of, for example, Bifidobacterium and Lactobacillus, while whey impairs abundance of Bacteroides fragilis and Clostridium perfringens in humans.Bacteroides, Alistipes and Bilophila increased following consumption of animal-based protein.Escherichia, Streptococcus and Enterococcus.Lactobacillaceae and Leuconostraceae abundance.Dietary proteins are derived from plants and animals. Several culture-based studies demonstrated that consumption of whey and pea protein extracts facilitates growth of Bacteroides and reduced Lactobacillus spp),Proteobacteria and Escherichia coli.Porphyromonadaceae spp in faeces of P80 fed mice.Bacteroides ovatus and Enterococcus faecalis).Food additives preserve a food product  or enhance the taste of processed food. Emerging evidence indicates that the consumption of food additives perturbs microbial composition and promotes experimental gut inflammation . For example, artificial sweeteners such as saccharin promote dysbiosis in mice , and IBD-associated dysbiosis exerts inflammatory functions in genetically susceptible mice. However, several aspects are poorly resolved in this context. For example, what is the specific impact of blooming pathobionts or loss of symbionts in human IBD and can this be therapeutically exploited? Critical mechanistic insights are probably best exemplified by studies on adherent invasive Experimental, translational and clinical evidence suggest that IBD arises from unresolved perturbation of mucosal immune responses that is determined by genetic variation and the exposome (including the diet and gut microbiota). This concept implies that a variety of cues, rather than a single incident, promotes the development of chronic unresolved gut inflammation, which may explain heterogenous results of key dietary intervention trials , which are summarised in EEN takes advantage of an elemental (liquid) diet that meets all nutritional demands of macronutrients and micronutrients and thus allows replacing (solid) dietary habits. There are plenty of formulas available, which greatly vary in their composition of macronutrients and micronutrients.EEN is the recommended first-line therapy in children and adolescents with active  mild-to-moderate CD that is usually used for 6–8 weeks,In adult patients with mild-to-moderate active CD, only few small studies suggest efficacy of EEN,A study in paediatric CD published in 2006 indicated that unrestricted partial enteral nutrition in combination with an elemental formula was less effective in inducing remission than EEN.In active mild-to-moderate UC patients that were refractory to therapy , a blinded, randomised, controlled trial with 62 participants from Israel and Italy investigated whether an exclusion diet with or without faecal microbiota transplantation could be effective. The UCED required dietary counselling that recommended enrichment of fruits and vegetables and disallowed Western dietary habits . The study was terminated early because the primary hypothesis that such a dietary approach would be beneficial in combination with faecal microbiota transplantation was rejected. However, the restriction diet alone induced clinical remission in 40% and endoscopic remission in 26% of patients at week 8 in this therapy refractory cohort,Collectively, these early clinical trials provide evidence that the diet impacts gut inflammation in mild-to-moderate CD and UC, and real-world experience suggested that dietary approaches are efficacious beyond clinical trials.SCD is a restrictive grain-free diet which claims to maintain remission in patients with IBD. The diet allows digestible monosaccharide carbohydrates, which are made of a single molecule and easily to be broken down without enzyme participation, for instance contained in fruits, nuts, eggs, most (non-starchy) vegetables, non-processed meat and fish, while complex carbohydrates derived from grains, corn, milk and cream and artificial sweeteners are restricted. In a survey of 50 quiescent IBD subjects who employed an SCD for 10 months, complete symptom resolution by self-report appeared to be 66%.Prevotella and fibre-degrading Firmicutes.The MD is rich in arguably healthy foods including vegetables, fruits, legumes, cereals, fish and unsaturated fats. Results from clinical and translational research on the MD point towards a use in managing IBD.Clostridium cluster XIVa and mucus-associated Akkermansia muciniphila.Fermentable oligosaccharides, disaccharides, monosaccharides and polyols (FODMAPs) are short-chain carbohydrates contained in wheat, onion, cabbage, legumes and stone fruits that are poorly absorbed in the small intestine. A diet low in these fermentable carbohydrates is called an LFD. In 89 adult patients with IBD  in remission or with mild-to-moderate disease, a randomised low FODMAP trial (vs a standard diet) for 6 weeks resulted in significant improvement in terms of quality of life and reduction of symptoms of concomitant irritable bowel syndrome.A GFD excludes all food items containing gluten, which is contained in wheat (and derivatives), barley, rye, triticale and brewer’s yeast, so that pasta, baked goods and beer (with other nuances) must be excluded from the diet. A cross-sectional questionnaire study with 1647 patients with IBD (with 0.6% concomitant coeliac disease or gluten-sensitivity) indicated that 20% have tried a GFD and that 66% of patients reported clinical improvement and 38% reported less flares.Preclinical and clinical studies from the last years demonstrated that the diet is a rheostat of microbial composition and function and may evoke dysbiosis, as exemplified by a human Western diet.


The appearance of chemoresistance in cancer is a major issue. The main barriers to conventional tumor chemotherapy are undesirable toxic effects and multidrug resistance. Cancer nanotherapeutics were developed to get around the drawbacks of conventional chemotherapy. Through clinical evaluation of thoughtfully developed nano delivery systems, cancer nanotherapeutics have recently offered unmatched potential to comprehend and combat drug resistance and toxicity. In different design approaches, including passive targeting, active targeting, nanomedicine, and multimodal nanomedicine combination therapy, were successful in treating cancer in this situation. Even though cancer nanotherapy has achieved considerable technological development, tumor biology complexity and heterogeneity and a lack of full knowledge of nano-bio interactions remain important hurdles to future clinical translation and commercialization. The recent developments and advancements in cancer nanotherapeutics utilizing a wide variety of nanomaterial-based platforms to overcome cancer treatment resistance are covered in this article. Additionally, an evaluation of different nanotherapeutics-based approaches to cancer treatment, such as tumor microenvironment targeted techniques, sophisticated delivery methods for the precise targeting of cancer stem cells, as well as an update on clinical studies are discussed. Lastly, the potential for cancer nanotherapeutics to overcome tumor relapse and the therapeutic effects and targeted efficacies of modern nanosystems are analyzed. During the last decade, cancer immunotherapy is a promising tool in the battle against cancer. Tumor immunotherapy, in contrast to conventional treatments, primarily targets immune cells. It stimulates the body’s immune response by preventing harmful immune regulatory factors and improving immune cells’ capacity to identify tumor cell surface antigens and eradicate tumor cells . AccordiCompared with the traditional chemo/immunotherapy scheme, modern nanocarriers display several advantages like increased bioavailability, lower toxicity, improved solubility and enhanced targeting , 18. SinCancer nanotherapeutics can be employed for the efficient targeting of tumor cells enabling passive targeted drug delivery Figure 2To accomplish the intended treatment outcome, controlled drug delivery systems (DDS) distribute therapeutic compounds to specified target cells, tissues, or organs in a pre-designed and controllable manner . TraditiThe tumor microenvironment (TME) is a key factor in the development of tumor heterogeneity and disease pathology , 48. DruIn photothermal therapy (PTT), cancer cells are killed by the created heat of exposing photothermal agents to light. Due to its noninvasiveness, high spatiotemporal precision, ease of usage, and flexibility of light sources, PTT has demonstrated considerable potential for the treatment of cancer Figure 4+T cells that infiltrated tumors while decreasing the percentage of regulatory T cells Doxil treatment boosted mature dendritic cells’ expression of CD80 in the tumor indicating that Doxil may cause these tumor-infiltrating cells to develop a costimulatory phenotype that can stimulate an anticancer T-cell response. Recently, PTEN-negative glioblastoma (GBM) is eradicated with anti-PD1 therapy assisted by Epirubicin-loaded nanosomes. By converting cold GBM into hot tumors with high infiltration of antitumor immune cells through the induction of ICD, removal of immunosuppressive MSDCs, and reduction of PD-L1 expression on tumor cells, the combination of epirubicin-loaded micelles with aPD1 eliminated GBM resistance to ICI (Immunogenic cell death (ICD), also known as immunogenic apoptosis, is a type of cell death that triggers a controlled immunological response. ICD promotes antitumor immune signaling against a variety of solid tumors in addition to directly killing tumor cells. Such methods for producing vaccine-like properties could be utilized to encourage an immunogenic, “hot” tumor microenvironment to develop from a “cold” tumor milieu, increasing patient response rates and producing positive therapeutic effects. ICD is characterized by the release of TAAs, DAMPs, and pro-inflammatory cytokines that make it easier for TAAs to be presented to adaptive immune cells and trigger an immunological reaction specific to an antigen. For example, in syngeneic animal models, Doxil enhances antitumor responses by cooperating with cancer immunotherapies . When coe to ICI .+ cancer stem-like cells residing in 3D mammary tumor spheroids  that can be injected or sprayed directly onto surface tumors after surgery or as a primary therapy. Once used, SFH has the ability to adjust shape in response to changes in tissue morphology that might take place during surgery. Nanoparticles made of microRNA (miRNA-215 and miRNA-206) and intrinsically disordered peptides are released by implanted SFH and infiltrate cancer cells to lessen their oncogenic hallmark. Four preclinical models of mesothelioma respond to a single application of SFH, suggesting the therapeutic value of local tumor-specific microRNA therapy  with an average diameter of 30-150 nm that are released by a wide range of cells throughout the body , 80. Thenication . Proteinigations .Through igations . Exosomeigations Figure 5igations . Exosomeigations . Furtherigations . Exosomeigations . In a diigations . Finallyin vivo tumor growth suppression  constitute a reliable and potent nanotherapeutic strategy. The maximal drug loading capacity, regulated drug release kinetics, longer blood circulation, and preferential tumor accumulation based on the EPR effect are only a few of the many inherent benefits of SAPs . These ppression . In addipression . In a disistance . Likewissistance . Recentlsistance .Nanogels are hydrogels with a particle size between 20-250 nm and have a 3D permeable structure . NanogelDrug delivery in the proper amount, at the proper time, and at the proper spot could considerably improve its efficacy in cancer treatment , 108. Th+T/Treg and CD8+T/Treg ratios (via the CD163 surface receptor and bind to endogenous plasma haptoglobin (Hp), thereby killing tumor cells. Additionally, the O2 generated by the Hb reduces tumor hypoxia, which strengthens the immune response against the tumor by deterring the recruitment of M2-type macrophages. The TME is synergistically reprogrammed by TAM-targeting depletion and hypoxia relief, which simultaneously reduces tumor cell PD-L1 expression, decreases the levels of immunosuppressive cytokines like IL-10 and TGF-b, increases the expression of immunostimulatory IFN-γ, improves cytotoxic T lymphocyte (CTL) response, and triggers a strong memory response  that can be used in order to boost anticancer immune responses , 70. Nang ratios . Also, tg ratios , 119. Furesponse . Tumor aresponse .Cancer nanotherapeutics have demonstrated a promising therapeutic alternative to current chemotherapy/immunotherapy regimens for overcoming or reversing drug resistance. Before being employed in clinical trials, these agents need to be better characterized and optimized. A better understanding of the tumor microenvironment, and development of novel approaches , and clinical trials utilizing nanosome-based systems are all suggested by the rapid development of nanotechnology and materials science. Utilizing nanotechnology’s advantages in tumor immunotherapy can successfully prevent drugs from unintentional deterioration and aid in achieving long-term circulation in both the blood and target tumor sites. The structure of nanoparticles needs to be changed in order to overcome the many issues restricting the present tumor immunotherapy approaches. Additionally, accurate immune response activation and immune suppression relief depend on an understanding of the immunotherapy’s controllable time and space. Thus, there is a great potential for therapeutic efficacy when tumor immunotherapy is combined with nanotechnology. In addition, significant consideration should be given to large-scale, reproducible commercial batches of nanomedicine formulations with improved efficacy and decreased toxicity. To achieve quick clinical conversion, a number of important factors need to be taken into account. First, by controlling the tumor microenvironment and metabolism, future research on metabolic pathways and cancer should be focused on enhancing cancer immunotherapy. Second, in order to specifically respond to the tumor microenvironment, nano-delivery devices with imaging capabilities and stimulus activation properties must be developed. Notably, tumor immunotherapy should be integrated into the development of nanomedicine’s diagnosis and treatment plans.Third, interdisciplinary research is required to find new immunological targets and pathways that will make it easier to create new drug delivery methods. This research should involve experts from pharmacology, materials science, immunology, and other domains. As a whole, the development of nanomedicine systems should concentrate on both commercialization and their ability to reach the clinical context.All authors contributed equally to the writing of this manuscript. All authors provided critical feedback and helped shape the manuscript. All authors contributed to the article and approved the submitted version.This research is co-financed by the ERAPEDMED/PMT-LC grant —Personalized multimodal therapies for the treatment of lung cancer (ERAPERMED2020-342) funded by the European network grant and the Greek General Secretariat for Research and Innovation (GSRT).The author would like to thank Professor Konstantinos Zarogoulidis for reading the manuscript and providing useful comments and advice.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


During their final life decade, people in Germany tend to live in senior residences or nursing homes. Re-location to such institutionalized homes is associated with declining levels of physical activity. Beside intrapersonal factors, like dealing with illness, social and physical environments might be decisive in shaping residents' physical activity levels. At the same time, it remains largely underresearched what residents themselves perceive as barriers and promoters of physical activity. This presentation aims to introduce first results of BaSAlt , a participatory research project on physical activity promotion in senior residences in Germany.We draw upon data from the BaSAlt study (2019-2022) in eight German senior residences, generated using Photovoice with 48 residents, staff, and visitors, as well as semi-structured interviews (n = 24), group discussions (n = 8), cooperative planning in future workshops (n = 16) and document analysis. Recorded and transcribed interviews, photos, discussions, as well as documents are subject to thematic analysis (Braun & Clarke 2016). All participating actors become part of data collection, and analysis, as well as in developing and implementing change projects to promote physical activity informed by findings.The presentation will highlight similarities and differences in participants' perceptions of physical activity-related promoters and barriers. We will present a preliminary comparative assessment of the organizational readiness in participating senior residences and potential consequences for cooperative planning in future workshops. Our aim is to initiate concrete action to improve physical activity promotion.The BaSAlt design recognizes residents, staff, and visitors as experts of their own lives, which allows for greater awareness of people's unique experiences and perceptions. The participatory approach used both for data collection and intervention planning might help uncover physical activity-related promoters and barriers that might otherwise have remained hidden and thus support the development of individualized and targeted action for physical activity promotion.


Reported prevalence rates of medically unexplained symptoms (MUS) in people aged ≥65 years range between 1.5 and 18%. People with MUS often describe a low quality of life and frequently suffer from co-morbid anxiety and depressive disorders. In our pilot study on older patients with MUS, the level of somatic comorbidity as well as frailty parameters were significantly higher among patients with MUS which was partially explained by a somatic origin compared to patients with MUS for which no explanation at all was found.The objective of this study was to examine the level of frailty and somatic comorbidity in older patients with medically unexplained symptoms (MUS) and compare this to patients with medically explained symptoms (MES).Frailty was assessed according to Fried’s criteria , somatic comorbidity according to the self-report Charlson Comorbidity Index and the number of prescribed medications.Although MUS-patients had less physical comorbidity compared to MES-patients, they were prescribed the same number of medications. Moreover, MUS-patients were more often frail compared to MES-patients. Among MUS-patients, physical frailty was associated with the severity of unexplained symptoms, the level of hypochondriacal beliefs, and the level of somatisation.Despite a lower prevalence of overt somatic diseases, MUS-patients are more frail compared to older MES-patients. These results suggest that at least in some patients age-related phenomena might be erroneously classified as MUS, which may affect treatment strategy.No significant relationships.


NCC activity is modulated by a variety of hormones, but is also influenced by the extracellular K+ concentration. The putative “renal-K+ switch” mechanism is a relatively cohesive model that links dietary K+ intake to NCC activity, and may offer new targets for blood pressure control. However, a remaining hurdle for full acceptance of this model is the lack of human data to confirm molecular findings from animal models. Extracellular vesicles (EVs) have attracted attention from the scientific community due to their potential roles in intercellular communication, disease pathogenesis, drug delivery and as possible reservoirs of biomarkers. Urinary EVs (uEVs) are an excellent sample source for the study of physiology and pathology of renal, urothelial and prostate tissues, but the diverse origins of uEVs and their dynamic molecular composition present both methodological and data interpretation challenges. This review provides a brief overview of the state-of-the-art, challenges and knowledge gaps in current uEV-based analyses, with a focus on the application of uEVs to study the “renal-K+ switch” and NCC regulation. We also provide recommendations regarding biospecimen handling, processing and reporting requirements to improve experimental reproducibility and interoperability towards the realisation of the potential of uEV-derived biomarkers in hypertension and clinical practice.The thiazide-sensitive sodium chloride cotransporter (NCC), expressed in the renal distal convoluted tubule, plays a major role in Na Howeverure (BP) –8. A recve years . These o+ handling has a profound effect on body fluid and BP maintenance as exemplified by the use of diuretic medications to treat states of volume expansion or hypertension. The thiazide-sensitive NaCl cotransporter  plays a key role in the regulation of Na+, Cl- and K+ balance and BP, as underscored by Gitelman syndrome (loss-of-function mutation in SLC12A3) resulting in normal to low blood pressure, hypokalaemia and metabolic alkalosis, combined with significant hypomagnesaemia and hypocalciuria. Activation of the renin-angiotensin-aldosterone system (RAAS) was initially thought to be a primary driver of NCC activity, but this system could not easily explain the observations that NCC activity decreased during high aldosterone states subsequent to high dietary K+ intake  increased NCC activity  are small vesicles derived from their cells of origin and have been detected in blood, urine and other biofluids. Urine is a favorable specimen for biomarker discovery and is used to diagnose and monitor kidney diseases. Proteome analyses reveal that EVs isolated from urine may represent a more targeted approach for biomarker discovery than unfractionated urine –20. EVs gulation . Therefo+ switch” and NCC regulation. We also provide recommendations regarding biospecimen handling, processing and reporting requirements to improve experimental reproducibility and interoperability towards the realization of the potential of uEV-derived biomarkers in hypertension and clinical practice.Urinary extracellular vesicles (uEVs) are a heterogenous group of nanosized membrane vesicles excreted by cells lining the urinary tract  that funEVs consist of exosomes, microvesicles and apoptotic bodies Figure 1 , Na-K-2Cl cotransporter 2 (NKCC2) and NCC  and a prominent population of small non-coding RNAs. mRNAs in uEVs derive from all regions of the nephron and could facilitate the examination of kidney cellular transcriptional changes in health and disease . The smaLipids and metabolites are relatively less investigated uEV components. However, in human uEVs over 100 lipid species have been detected by mass spectrometry, with cholesterol the most abundant lipid species, followed by phosphatidyl serine . CompareThe general steps for studying uEVs include pre-analytical urine handling, uEV isolation, uEV characterisation and normalisation, and downstream analyses of uEV content. Careful consideration of the applied technologies and approaches used at each step are necessary as these aspects are major source of data variability and can limit data reproducibility , 78–80.Pre-analytical urine handling is a critical source of variability as preservation and storage methods have a major impact on outcomes. Important steps in urine handling include urine collection , processing and storage , and urine handling after thawing . Huebner and colleagues summarized and compared currently applied methods for urine collection and storage for uEV utilization . Their sUltracentrifugation was the first method used to reproducibly isolate uEVs . HoweverNew methodologies used for uEV isolation include EV filtration techniques, uEV precipitation, hydrostatic dialysis, acoustic trapping and immunocapture Table 2.EV characterisation is not straightforward, because non-EV entities, such as argonaut 2 protein complex and lipoproteins also contain components that are present in EVs , 105, anUnfortunately, there is no single technique that can fulfill the requirements for describing EV morphology, size, number and content. Current techniques used for EV morphological characterisation include transmission electron microscopy (TEM), cryogenic electron microscopy (cryo-EM), atomic force microscopy (AFM) and super resolution fluorescence microscopy, among which TEM is the most commonly used. Because these techniques provide different information about EV structure and size distribution, they are not necessarily interchangeable or capable of providing images of comparable quality. Techniques used to measure uEV size distribution and count include nanoparticle tracking analysis (NTA) and tunable resistive pulse sensing (TRPS). On the basis of Brownian motion, NTA provides uEV particle size distribution and concentration within a specific detection range, however it cannot exclude non-EV entities and therefore NTA-based particle count may be overestimated. TRPS is a relatively more accurate method that measures particle size, number and surface charge . ComplemOne of the greatest challenges in uEV-based research is the requirement for normalisation of uEV contents between samples or subjects, especially in clinical studies where spot urine is the most frequently used and simplest collection method. Current normalisation approaches, each with both benefits and limitations, include comparing uEV contents to THP abundance, urine creatinine concentration, known EV-enriched proteins, uEV concentration or uEV numbers. THP abundance highly correlates with EV markers such as ALG-2-interacting protein X (ALIX) and Tumor Susceptibility Gene 101 Protein (TSG101) , but theUrine creatinine is widely used to normalize analytes (e.g. urine protein-to-creatinine ratio) in spot urine samples in routine clinical practice , but itsThe abundance of EV-enriched proteins and uEV concentration are considered relatively stable . As suchThe uEV concentration depends not only on the isolation method used, but also the uEV production and excretion rate (secretion minus possible uptake) and the overall urine concentration at the sampling time point. uEV excretion has showed a circadian pattern that is independent of sex, fed-fasting/hydration status and kidney injury, and this circadian variation can be normalised by uEV particle number and its marker protein TSG101 . A more Measurement of uEV protein abundances is frequently used to reflect ongoing changes in the kidney or identify biomarkers for specific diseases. For example, uEVs have been used for quantitative assessment of the levels of different proteins in the setting of patients with familial hyperkalaemic hypertension , Bartter+ reabsorption and K+ excretion, and these processes are intimately related, as exemplified by amiloride induced hyperkalaemia and the presence of hypokalaemia in patients with thiazide induced hyponatraemia. The primary channels involved in distal tubular Na+ handling include ENaC expressed from the late distal convoluted tubule (DCT) through the connecting tubule (CNT) and the medullary collecting duct (CD), and NCC expressed only in the DCT. ENaC is thought to be the major target of aldosterone in the distal nephron. Although NCC activity was originally proposed to be increased by aldosterone, more recent studies indicate this largely occurs through low K+ induced increases in NCC phosphorylation (which activates the transporter) via activation of the Cl‐ -sensitive With-No-Lysine kinases (WNK) and SPS1-related proline/alanine-rich kinase (SPAK) network  K+ concentrations  of intravenous NaCl loading alone on NCC in patients with primary aldosteronism was recently examined  than those with non-suppressed RAAS, with inverse correlations of plasma K+ and uEV abundances of NCC and pNCC. These observations are similar to patients with primary aldosteronism, that uEV levels of pNCC are indeed a reliable readout for NCC activity and that plasma K+ may override any stimulatory effect of mineralocorticoid/aldosterone on NCC. The hypokalaemia in patients with primary aldosteronism or Cushing’s syndrome may contribute to hypertension through increased Na+ reabsorption via enhanced NCC activity. Recently, Kong et al. reported that differences in NCC and pNCC in uEVs can be used as biomarker for subtyping of primary aldosteronism and potentially for avoiding the need for adrenal venous sampling (AVS) in differentiating unilateral from bilateral forms .BioRender.com.Cartoons in Figures were created with The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


With rich carboxyl groups in the side chain, biodegradable polymalic acid (PMLA) is an ideal delivery platform for multifunctional purposes, including imaging diagnosis and targeting therapy. This polymeric material can be obtained via chemical synthesis, or biological production where L-malic acids are polymerized in the presence of PMLA synthetase inside a variety of microorganisms. Fermentative methods have been employed to produce PMLAs from biological sources, and analytical assessments have been established to characterize this natural biopolymer. Further functionalized, PMLA serves as a versatile carrier of pharmaceutically active molecules at nano scale. In this review, we first delineate biosynthesis of PMLA in different microorganisms and compare with its chemical synthesis. We then introduce the biodegradation mechanism PMLA, its upscaled bioproduction together with characterization. After discussing advantages and disadvantages of PMLA as a suitable delivery carrier, and strategies used to functionalize PMLA for disease diagnosis and therapy, we finally summarize the current challenges in the biomedical applications of PMLA and envisage the future role of PMLA in clinical nanomedicine. The biosynthesis of polymalic acid (PMLA) and its biotechnical high-grade production from microorganisms compared with the chemical synthesis of PMLAThe physicochemical and biological characteristics of PMLA and its derivativesHow PMLA’s general chemical characteristics can be used to generate various macromolecular compounds for pharmaceutical deliveryThe concepts of biological and clinical targeting exemplified by PMLA-based drugs and imaging agents and their biodistribution and biodegradabilityAn evaluation of the mechanisms that generate preclinical antitumor efficacy and the translational potential for clinical imaging Similarenolysis –25. The ivatives –28.Synthetic PMLAs from the aforementioned chemical reactions have been evaluated for biomedical applications. For instance, PMLAs have been used as fusogenic materials with artificial phospholipid bilayers  to enhance their thermo- or pH- sensitivities in medicinal uses , 30. Howl-malic acid) from sustainable sources through one-pot fermentation, revealing the engineerable potential of biosynthetic PMLAs . Th. Th13].  lengths , 39.Fig.P. polycephalum, PMLA is only synthesized at the plasmodium stage in the nuclei of multinucleate coenocytes. PMLA has not been found in any of its mononucleate forms in the life cycle of P. polycephalum; however, the synthetic rate of PMLA production increases in parallel to mitotic nuclei enlargement, evidencing the indispensable role of PMLA in maintaining synchronous functioning .Fi.FiP. polMw = 50 kDa), together with LLL to facilitate an endosome escape and rhodamine (rh) for fluorescent tracing [Recently, by switching the navigator antibody in Polycefin, a BBB-penetrating peptide Angiopep-2 (AP-2) was chemically attached onto the PMLA base  or are bound to electrolytes of opposite charge (via cationic complexation). The PMLA-based nanostructures obtained are thus restabilized as nanoscale platforms, enabling the loading and controlled release of pharmaceutical molecules.Mw = 34 kDa) in acetone were mixed for the reactions, and partial esterification of the pendant carboxylates was accomplished at random [Methylated PMLAs were obtained in a typical one-pot synthesis in which different ratios of diazomethane over PMLA (t random . In aquet random . Hydropht random . This drKa = 8.6) were added into aqueous solutions of PMLA with a fixed mole . The increasing amount of DOX made the solution change from colloidal suspension to reddish precipitation, and > 90% added DOX formed ionic complexes with carboxylic acids of PMLA via their amines [Polymalic acid under most biological conditions becomes a polyanion that possesses a number of carboxylate ions, which can easily interact with cationic moieties to form stable complexes . In a tyr amines . After Dr amines . The loar amines .Fig. 9PMnATMA, where n represents the number of carbon atoms in the alkyl chain) were chosen to form ionic complexes with PMLA (CnATMA-PMLA). The ionic complexes were studied to determine their interplay with antibiotics for drug loading and release [nATMA, helps to prevent a water/enzyme attack that would break down the ester bonds in PMLA backbone, resulting in a slower hydrolytic or enzymatic degradation of CnATMA-PMLA and consequently releasing single units of erythromycin-bound malates at a slower rate  [Alternatively, anionic PMLA can first react with cationic compounds to form a supramolecular assembly where its property of loading and releasing drugs becomes further adjustable by choosing a variety of cationic counterparts. In a pilot study, alkyltrimethylammonium (ATMA) surfactants with different lengths of alkyl chains  .Mw (205.4 kDa) produced from the liquid fermentation of Aureobasidium pullulans var. pullulans MCW, was mixed with cysteine-conjugated chitosan (Cys-CS) via ionic gelation at different weight ratios to form particles of different sizes and surface charges [Helicobacter pylori). These particles were chosen to load antibacterial amoxicillin, further showing a pH-triggered release to inhibit the growth of Helicobacter pylori while leaving stomach cells unharmed , and their film growth exhibited a high degree of linearity between film thickness and the number of tetralayers. This tetralayer architecture demonstrated an exceptional diffusiveness between the layers of film, which would be advantageous for cargo release [n tetralayer prevented the abrupt release of biologics in the case of (protein/PMLA)n bilayers, sustaining a timed release due to the hydrolytic degradation of PMLA and the accompanied dissociation of ionic complexes in response to changes in environmental pH and ionic strength; comparatively, dissembled chitosan could preserve the activity of the released proteins by preventing them from being denatured or proteolyzed [In addition, a biodegradable thin film was prepared after a layer-by-layer assembly of PMLA and chitosan, revealing an excellent capacity for the controlled release of proteins, including lysozyme and basic fibroblast growth factor (bFGF) . To inco release . It was teolyzed .Mw  or first interacts with oppositely charged polyelectrolytes of high Mw and then with functional compounds, the complexes undergo a completion of reactions in aqueous solutions. These series of reactions aid in unleashing more absorbed therapeutic or diagnostic reagents with the addition of stimuli or by stimuli-triggered self-degradation of PMLA main chain compared with free drug diffusion. Accordingly, PMLA hydrolysis, together with the dissociation of the ionic complex, liberates the drug-bound malate units.In PMLA-based ionic complexation, whether PMLA forms stable complexes directly with positively-charged functional compounds of low l-malates. These products can be further utilized in the TCA cycle and metabolized into CO2 and water, constituting no secondary harm to the body or no residual deposition in the body.Here we review the recent advances in the chemical synthesis and bioproduction of PMLAs, their physicochemical and physiochemical characteristics, their biomedical applications as delivery platforms for disease diagnosis and treatment, and the unique chemical methods used to engineer PMLA-based nanomaterials. Produced from biological sources rather than chemical reactions, PMLA has several advantages as a unique platform for theranostic delivery in the clinical translation. Firstly, manufactured through fermentation engineering of microorganisms, PMLA is a biorenewable natural product with promising full-scale industrialization. Secondly, water solubility of PMLA is unrivaled when compared with water-insoluble biopolymers, such as cellulose, polyhydroxybutyrate, etc., or biopolymers that are only water soluble in salt form, such as polyglutamic acid. More importantly, this excellent water solubility is inherited by many PMLA-derived complexes, maintaining their suitability as delivery platforms. Thirdly, pendant carboxylic acids along the main chain of PMLA offer several opportunities for further covalent conjugation or ionic complexation, making PMLA a resourceful carrier for therapeutic and diagnostic delivery. Fourthly, the release mechanism of a PMLA carrier is mostly due to the hydrolytic breakage of the ester bonds in its backbone, although it could be complemented with the dissociation of other intermolecular forces, such as ionic bonds. The timing of this release profile greatly avoids a premature cargo leakage before delivery to the diseased lesion. The degradative or hydrolytic rate of PMLA under physiological conditions could be favored by pharmacokinetics. Lastly, the full biodegradability can make PMLA adaptable to many formulations, including orally, by injection, or even through implantable medical devices, as the final degradation products comprise of sole However, after decades of PMLA research and technological development, challenges still remain. The first problem lies in the PMLA’s unscalable production and functionalization for medicinal purposes. Following current manufacturing flow and purification processes, the regular yield of bioproduced PMLA remains on a scale of grams per batch, taking up to two weeks per production cycle . FurtherMw of PMLA cannot be controlled during its biosynthesis, so it is impossible to tailor the length of bioproduced PMLA as desired. In parallel, the relationship between the Mw of PMLA and its suitability as a delivery platform also needs to be determined. The higher the Mw of PMLA, the greater its loading capacity and the longer its delivery time is before its hydrolytic degradation; however, this relationship has not been quantitatively defined or precisely assessed.The second challenge remains that the genes responsible for PMLA production in several microorganisms are yet to be fully uncovered. Once identified, these genes could be introduced into the commonly used industrial microbes for the biosynthesis of this precious natural product using metabolic engineering, thereby largely lowering the manufacture cost, boosting the yield, and commercializing the product. For this reason, the Last but not least, there are unexplored opportunities in which PMLA could be used for more applications other than a delivery platform. With unrivaled hydrophilicity, PMLA-based drug delivery can be applied to many fields insofar uncharted, such as kidney diseases and urological cancers/infections. Besides, as a fully degradable biopolymer, PMLA could be useful in fields other than biomedical research, such as the food or pharmaceutical industry for external packaging. Its applicability to these fields would only help promote research into its medicinal development.In closing, PMLA is currently one of remarkable materials for many biomedical applications, including pharmaceutical delivery and tissue engineering. Its nontoxic, non-immunogenic, and fully degradable features make it a perfect medicinal platform, holding great potential in targeted biomolecular delivery and many other pharmaceutical and biological applications.


Oculomics, is a growing field of investigation for early diagnostics, enabling structural and chemical biomarkers to be monitored overtime to predict prognosis. Traumatic brain injury (TBI) triggers a cascade of events harmful to the brain, which can lead to neurodegeneration. TBI, termed the “silent epidemic” is becoming a leading cause of death and disability worldwide. There is currently no effective diagnostic tool for TBI, and yet, early-intervention is known to considerably shorten hospital stays, improve outcomes, fasten neurological recovery and lower mortality rates, highlighting the unmet need for techniques capable of rapid and accurate point-of-care diagnostics, implemented in the earliest stages. This review focuses on the latest advances in the main neuropathophysiological responses and the achievements and shortfalls of TBI diagnostic methods. Validated and emerging TBI-indicative biomarkers are outlined and linked to ocular neuro-disorders. Methods detecting structural and chemical ocular responses to TBI are categorised along with prospective chemical and physical sensing techniques. Particular attention is drawn to the potential of Raman spectroscopy as a non-invasive sensing of neurological molecular signatures in the ocular projections of the brain, laying the platform for the first tangible path towards alternative point-of-care diagnostic technologies for TBIThe study of ocular manifestations of neurodegenerative disorders,  I.Traumatic Brain Injury (TBI) can cause death or lifelong physical and mental disability. TBI occurs when the brain is damaged by rapid acceleration or deceleration with rotational or shear forces or penetration While there are numerous extensive reviews of ocular manifestations of common neurodegenerative (ND) diseases A.TBI may occur as a result of closed, open or crush head injuries, disturbing brain function. Most commonly, there is no break to the skin and the brain accelerates and/or decelerates within the closed skull, twisting, stretching and damaging the axons and blood vessels. Less often, the brain is exposed by an open injury whilst the skull base and brain stem may be damaged by crush injury The most common brain areas involved in TBI are frontal and temporal lobes Moderate to severe TBI as associated with hematomas (blood leakage) and immediate tissue death, not only losing brain functionality but also releasing toxins TBI has been termed a “silent epidemic”, as many incidents go unreported by patients or unrecognised by healthcare professionals B.TBI pathophysiology can be split into acute, sub-acute and chronic phases, occurring within 3-24 hours, 1 day - 3 weeks and from 3 weeks of the trauma, respectively Most established TBI diagnostic techniques can only be implemented in hospital, and a patient would typically undergo multiple examinations before a final diagnosis is made. Out-of-hospital assessment of TBI severity includes the Glasgow Coma Scale (GCS), a neurological scoring system of the patient's conscious state. Neuroimaging is usually only available in a hospital setting, as are surgical interventions such as decompression and insertion of monitoring devices 1)The National Institute for Health and Care Excellence (NICE) highlighted the need for a test to allow paramedics to determine if the patient should be transported from the scene of injury directly to the nearest neurological centre or a hospital with particular resuscitation resources Even in the absence of confounding factors, the prognostic accuracy of the on-site GCS assessment is lower than GCS on admission to hospital 2)Other current early diagnostic techniques include neuroimaging and ICP monitoring, which are often equivocal on the presence and severity of TBI, particularly in the mild and moderate groups, whilst also being costly, slow, time consuming and requiring highly trained personnel to perform and interpret Neuroimaging is not often performed independently as it can provide false-negative results, for example 29% of TBI patients with negative CT scans show positive MRI findings 3)Current TBI diagnostic pathways frequently result in over-diagnosis and over-triage of TBI, which creates high healthcare costs from the avoidable tests and treatments Mild TBI cases are the most difficult to identify and only 5% of TBI cases in U.K. emergency departments each year are moderate or severe injuries based on GCS categorisation TBI severity is often variably defined because of injury heterogeneity. Mild TBI can accompany one or multiple symptoms of headaches, nausea and disorientation, lasting from days to weeks Many cases of TBI, particularly mild TBI, cause functional and metabolic abnormalities without any detectable structural damage, or damage at a molecular level that is un-detected by neuroimaging Moderate and severe TBIs, on the other hand, are accompanied by more obvious and immediate symptoms indicating neurological injury. However, in the context of polytrauma and substance abuse, which may also affect conscious level, this does not guarantee correct identification of TBI or categorisation between mild and severe injuries, especially as both severity levels may coincide with initial unconsciousness Variation in presentation generates different drivers for POC diagnostic techniques. Cognitive techniques fall short for all severities due to mild or delayed effect on mild TBI patients, whilst moderate-to-severe patients experience greater time spans of unconsciousness. Mild cases would benefit from techniques that utilise biochemical responses, as structural changes are less common, whilst early structural changes do not effectively differentiate moderate-to-severe cases, and thus sensitive and specific biochemical responses may aid differentiation. Thus, POC TBI diagnostics would benefit from a sensitive biomarker imaging technique capable of categorising all severity levels. Monitoring the biochemical and metabolic changes over time following initial injury in the acute phase  and eye movement, facial muscles and taste  , or dysfA.Many of the areas of the brain most vulnerable to mild TBI are related to vision B.After TBI, axonal damage in both the ON and retina is indicated by ON oedema caused by stretching and oxidative stress et al. demonstrated in a murine model that TBI biomarkers may be transported from the brain to blood via the CSF in the glymphatic system et al. explored mild TBI in rats, observing an increase in glymphatic influx and decrease in glymphatic efflux, resulting in CSF being inadequately cleared, contributing to cerebral edema associated with TBI CSF is present in subarachnoid space surrounding the ON and involved in the ocular glymphatic system There has also been a recent increase in research surrounding Spaceflight-Associated Neuro-ocular Syndrome (SANS), which is a series of ophthalmologic and neurologic findings in astronauts following spaceflight. SANS encompasses ON head swelling, globe flattening, choroidal/retina folds and increased CSF volume in ON sheaths et al. analysed the potential compartmentalisation of the CSF after ON disorders and found that CSF is biochemically continuous up until the ON, but CSF obtained from the subarachnoid space of the ON differed from that surrounding the brain, with both fluids containing different biomarker concentrations et al. were able to detect beta-2-transferrin in the subconjunctival fluid with a 25-gauge needle, a marker of CSF leaks Killer C.RGC loss occurs alongside ON degeneration as early as seven days after mild TBI and persist at 6 months post-injury, detectable using OCT both in human and animal studies Many systemic and cerebral neurodegenerative (ND) diseases manifest in the eye including Alzheimer's disease (AD), a form of dementia most commonly associated with memory disturbance, motor neuron disease (leading to muscle weakness), Parkinson's disease (PD), another form of dementia presenting as a movement disorder and Multiple Sclerosis (MS), a CNS disorder that causes vision, motor and sensory difficulties et al. linked clinical retinal diseases with dementia, including macular degeneration, glaucoma, and diabetic retinopathy, which develop from biochemical pathologies that could be identified even before cognitive decline and changes in vision become apparent Structural and physical ocular changes may be measured in the “early” phases of neurodegeneration (droopy eyelid) Eye movement and afferent visual abnormalities are amongst the earliest clinical manifestations of ND diseases, including PD D.in-vivo ocular imaging technique that utilises laser interferometry The relationship of changes in ocular structures, such as the retina, ON, choroid, pupil, and lens as well as tears, with neurodegenerative diseases has been extensively reviewed elsewhere et al. suggested that retinal amyloid-Beta tests can differentiate between the presence of AD with 100% sensitivity The retina provides a convenient window to assess neuronal and vascular changes in the CNS The retina and ON  can undeE.CT scans and ultrasound have identified ON sheath distention in all TBI severities, related to elevated ICP F.via the ON An inflammatory response within the brain can affect the retina Whilst most studies focus on structural changes, biochemical responses will develop much more rapidly than structural changes and may be detectable much earlier in the ND disorder pathogenesis III.Identifying chemical responses to TBI and correlation with injury severity would enable a better mechanistic understanding, development of new diagnostic modalities and opportunities for more effective therapies. Recent publications investigate chemical responses to TBI for early and accurate diagnostics NICE guidelines recommend focussing research on biomarkers for diagnosing, monitoring progression and stratifying patients with TBI for therapies A.et al., stated that ‘good’ biomarkers should be in easily accessible biofluids, with low background levels in healthy control groups, have high sensitivity and specificity to all injury severities and be released in a “time-locked sequence” after the injury Many TBI biomarkers are assayed in accessible biofluids Although well recognised and validated, acute TBI biomarkers may overlap with other disease and injury processes, such as extracranial plasma or serum, proteins released from damaged cells, polytraumatic organ and muscle injury Recognised biomarkers measured in the acute phase after TBI include, S100B, glial fibrillary acidic protein (GFAP), neuron-specific enolase (NSE), Ubiquitin C-terminal hydrolase-L1 (UCHL1), total tau (t-tau) protein and αII-spectrin breakdown products The most widely investigated acute biomarkers include S100Β, a calcium-binding protein expressed in glial cells of the CNS regulating calcium level, and GFAP, a structural, filament protein of astrocytes located in the CNS CSF, associated with astrocyte and BBB injury As S100B is widely researched and documented, the healthy baseline concentration in adult serum is stated and agreed to be approximately 0.11 ug/L GFAP is brain-specific and raised concentrations in the CSF Biomarkers associated with neuronal damage include NSE, a glycolytic enzyme and UCHL1, a protease expressed in red blood cells (RBCs) and CSF, respectively UCHL1 is a cytoplasmic enzyme found in a high abundance in neurons αII-spectrin is a cytoskeleton protein in axons and presynaptic terminals αII-spectrin protein degradation in response to necrosis (uncontrolled cell death) are a potential TBI biomarker for secondary injuries such as, ischaemia t-tau) protein is a microtubule-associated protein expressed in neurons and astrocytes t-tau levels are elevated at various injury severities Total tau (Micro RNA (miRNAs) are small molecules that contribute to gene expression regulation, blood, CSF and brain tissue levels Plasma S100B and GFAP remained elevated throughout the entire 24-hour period after TBI and were consistently more elevated in TBI patients with unfavourable outcomes  [199]. BB.et al. utilised microdialysis to establish elevated extracellular NAA levels after TBI due to an efflux of NAA due to adverse brain chemistry which then decreases in the 50-96 hour period post-injury et al., and illustrated in et al. demonstrated decreased NAA concentration in normal-appearing white matter after axonal damage, linking NAA levels with mitochondrial dysfunction and acute inflammatory lesions et al. found that in ischaemic rat brains, there was a continuous decrease in NAA levels in animals with tissue death N-Acetyl-Aspartate (NAA) is a highly abundant amino acid derivative in adult brain neurons The BBB is compromised in the acute phase after head injury with increased glutathione (GSH) levels C.Levels of GFAP and UCHL1 increased in plasma after direct trauma in a porcine ON crush model t-tau and NAA are potential candidates, since t-tau has been identified in the retina in AD t-tau, NAA, αII-spectrin, GSH and NSE are all expressed in or associated with neurons and axons Disproving CSF compartmentalisation would suggest the possibility of biomarker detection in the CSF such as, the GFAP IV.Developments in correlating TBI biomarker levels with trauma severities would constitute a major step towards more accurate identification and understanding of head injuries, particularly mild TBI or concussion in-vivo biofluids rapidly, effectively, and non-invasively. Urine is a potential ex-vivo biofluid for detection of brain biomarkers ex-vivo sampling incorporate additional risk of samples being affected by preparation and preservation methods as was illustrated by Abdelhak et al. who has freeze- thawed CSF samples over 5 cycles showing a 50% decrease of the GFAP within those These molecular sensing techniques all exhibit attributes as potential routes for biomarker detection and TBI diagnostics however currently, no single method has the capacity to measure Recently, the FDA approved the Banyan Trauma Indicator (BTI), a novel biomarker screen to aid triaging of mild to moderate TBI patients before undergoing CT scans, with the intent of reducing costs and exposure to radiation, along with increasing availability to other patients. The BTI measures the UCHL1 and GFAP levels in blood samples, indicating the presence of intracranial lesions in the form of positive or negative assay results In clinical settings, optical techniques are favoured for being highly sensitive, non-destructive and rapid. Optical brain imaging methods capable of identifying microscopic structure and function have been continuously developed over the past 40 years since Jöbsis first measured brain blood and tissue oxygenation using near-infrared light A.in-vivo measurements due to difficulties in maintaining physiological states of biological samples Mass spectrometry measures the mass-to-charge ratio of gas-phase ions, which involves ionising the sample to break down molecules into fragments of separate masses and analyse them in-vivo when combined with magnetic resonance spectroscopy and capable of non-destructive sample analysis, but has a lower sensitivity than mass spectrometry in-vivo applications and greatly limit use pre-hospital or to obtain real-time monitoring data Mass spectrometry is often compared to another metabolomic technique, nuclear magnetic resonance (NMR), which can be utilised B.in-vivo analytical technique that non-invasively interrogates brain tissue metabolism. Similar to MRI, radio-frequency electromagnetic radiation is used to excite nuclei into alignment and then stopped to allow nuclei to return to their original state, creating a fluctuating magnetic field which is detectable as a current induced in the receiver coil Magnetic resonance spectroscopy (MRS) is an via spectral peaks Proton MRS uses the radio-frequency signal from MRI to obtain a chemical shift from a sample, which indicates its metabolite concentration MRS holds great potential in the post-injury follow up period, when it may be useful for monitoring neurodegeneration, which can complicate the long term follow up of patients with TBI C.in-vivo imaging of NO brain concentrations in humans Fluorescence occurs when incident light undergoes the Stokes Shift, in which the scattered light has longer wavelengths than that of the source et al. have utilised fluorescent signals with confocal laser scanning ophthalmoscopy to single retinal nerve cell apoptosis in-vivo and termed the technique detection of apoptosing retinal cells (DARC) in-vivo in the acute phase of TBI to diagnose and characterise injury severity, but require numerous patient preparation steps, firstly being a single injection dose of ANX776 to visualise cells during imaging, followed by pupillary dilation.Cordiero et al. imaged human retinal pigment epithelium (RPE) cells from ex-vivo fovea, perifovea and near-periphery, using confocal fluorescence microscopy et al., but it does not so readily detect inner retinal  damage et al. utilised lipofuscin, the main source of autofluorescence in human fundus, in patients diagnosed with uveitis using FAF, demonstrating its potential as a non-invasive, single follow-up tool for progressive inflammatory disorders affecting the outer retina Bermond D.e.g., a halogen lamp, although supercontinuum lasers may also be used. The specific interactions of the light with the various chromophores in the tissue can result in reflected, emitted or fluorescent light from the sample, which are received by a detector Hyperspectral imaging (HSI) involves the acquisition of two-dimensional images across a broad range of the electromagnetic spectrum. The precise number of wavelengths varies in the literature and there is some overlap and often arbitrary differentiation from multispectral imaging (MSI). However, HSI is differentiated from MSI by primarily relying on the use of narrow adjacent spectral bands over a continuous range, as opposed to the discrete and spaced wavelength bands in MSI in-vivo human applications, due to complications introduced by complex imaging equipment and computational issues in-vivo brain metabolism and haemodynamic measurements, reviewed here HSI can provide real-time images of a sample and as such has application in many tissues and biological contexts. Regarding the brain, it has been used to assess cerebral blood flow and tissue oxygenation Multispectral imaging has also been developed towards ophthalmic applications, although again these are focussed on the outer retina. Histological parameters have been extracted from multispectral images of the human ocular fundus to map the concentration and distribution of the retinal haemoglobins, choroidal haemoglobins, choroidal melanin, RPE melanin and macular pigment E.in-vivo technique that has undergone considerable testing in human subjects Near-infrared spectroscopy (NIRS) is an NIRS cannot discriminate between arterial, capillaries or venous blood and therefore provides a combined reading of tissue blood oxygenation. As venous blood is most abundant within the cranium, the normal cerebral oxygenation value obtained by NIRS is lower than that obtained by pulse oximetry which assess the pulsatile arterial blood oxygenation signal et al. investigate and review the promising applications of NIRS in TBI monitoring, collating examples of experiments that find good agreement between NIRS and current gold standard techniques, i.e., neuroimaging, ICP monitoring Rodlan F.ex-vivo technique and requires tissue to be sectioned. Wang et al. identify THz spectroscopy as an early diagnostic tool for blast-induced TBI and able to differentiate the serum and CSF of mice with differing TBI severities Terahertz (THz) spectroscopy uses electromagnetic radiation in the wavelength range between microwaves and infrared to detect properties of matter. The vibrational and rotational energy of many biomolecules, such as proteins, are within this range thus making it an effective analytical technique that does not require pre-processing or labelling G.RS is a powerful, sensitive and specific technique capable of measuring the chemical composition of complex samples, which can be accomplished non-invasively in a label-free manner −1) against intensity (arbitrary units) Given that the intensity of a Raman signal is a millionth of the incident light source due to the low probability of inelastic scattering −1 due to C-C-O bond symmetric stretching vibrations along with two peaks at 1050cm−1 and 1090 cm−1, due to the C-O bond scaling and C-C-O bond stretching vibrations, respectively in-vitro neuronal cells ex-vivo using either biopsies or blood samples Raman spectroscopy (RS) has shown promise in neuro-applications and has been utilised to analyse neurochemistry ex-vivo characterisation of ND diseases, focussed on the study of blood and tissue in both animal and human models, with only a single human study of MS, in which Bergholt et al. investigated the lipid content of post-mortem brain tissue in-vivo capability for real-time imaging of an MS model RS also detected neurodegeneration, allowing early and sensitive neurodiagnostics through the rapid, non-invasive spectroscopic assessment of biofluids at the point-of-injury RS applications in AD diseases are more extensive and the research is majorly split into the characterisation of CSF or blood to identify protein and lipid biomarkers 2)RS is a non-invasive technique able to measure biomarkers rapidly and in real-time. As a molecular sensing technique, RS could be a powerful method for detecting TBI indicative biomarkers The multiplex ability to obtain chemical information and detect low-level biochemical changes in tissue samples makes it a potentially powerful technique in, not only diagnosing TBI but also in, deciphering between mild, moderate, and severe cases. RS applied in animal TBI injury models ex-vivo studies in detecting and monitoring TBI biomarkers in various biofluids TBI indicative biomarkers from human plasma have been characterised using surface enhanced Raman spectroscopy (SERS). One system demonstrates that combining an optofluidic, SERS lab-on-a-chip with a portable RS system presents promise for injury diagnostics and subsequent monitoring by profiling the levels of the NAA, S100B and GFAP V.A.ex-vivo Bruch's membrane samples, the inner-most layer of the choroid, suggesting the potential of doing the same in-vivo to determine those at high risk of ocular disease and the success of therapy and also that Bruch's membrane and sclera are composed of similar biomolecules, suggesting the sclera could be used as a viable (and more readily accessible) surrogate marker for changes in the Bruch's membrane RS analysis of tears and tear meibum have shown promise, studying composition Ex-vivo studies of enucleated human eyes have shown promise for development of ocular RS in-vivo studies et al. continued previous work in mouse retinae, identifying retinal layers using RS and discriminating between healthy and AD subjects en-face healthy and AD retinae are not distinguishable without statistical analysis to detect underlying changes  using an in-vivo RS measurements of the tissue, examining neuroinflammatory molecular changes as a function of time Stiebing  changes , which wB.In-vivo RS measurements of the lens has also been explored with the aim to identify the presence of artificial intraocular lenses used for the treatment of cataracts and investigated the effect of lens yellowing and transparency loss on in-vivo RS measurements et al. explored the prospect of monitoring inner nuclear layer thickening within the retina by coupling RS with a confocal scanning laser ophthalmoscope. The study measured key molecules of inflammation and neurodegeneration in human retina, demonstrating the potential of RS non-invasively addressing molecular changes of in-vivo CNS via the retina et al. provided a proof-of-concept design that used RRS to obtain in-vivo macular pigment measurements within the human retina, patented in 1999, limiting the majority of succeeding publications to the same research group and method In 1998, Gellermann in-vivo human retina, measuring the concentration of macular carotenoid pigments lutein and zeaxanthin  with a 488 nm excitation wavelength to create a strong vibrational response from the carotenoid, permitted a lower laser power and exposure time to be used compared to previous work and facilitated confocal Raman spectra of an axanthin  and obta results  [416]. Mex-vivo using the 785 nm excitation wavelength predicting the success of the future in-vivo RS system to diagnose macular degeneration in-vivoet al. have recently detected, distinguished and categorised the response of segments of an ex-vivo porcine eye samples using a 785 nm excitation wavelength  and often waste resource through over-triage. There is therefore a strong need for reliable, early, POC molecular diagnostics to support triage and clinical decision-making to allow patient treatment to improve outcomes within the golden hour.This paper has presented an overview of the status, opportunities, and obstacles faced as researchers begin to explore early diagnostic tests for TBI and other neurodegenerative diseases. The translation of neurological biomarkers from the bench to the secondary care and pre-hospital, POC arenas is progressing rapidly in a number of areas, showing promise particularly in spectroscopic techniques such as RS. To accelerate the field of biomarker imaging, categorisation is key. Clear standard ranges and concentration thresholds of TBI biomarker(s) in blood and CSF the acute phase after injury would accelerate diagnoses and severity characterisation for all biomarker detection techniques. Not only would this facilitate diagnoses but also aid better understanding of an incredibly heterogeneous pathology, stratifying biochemical responses to differing injury severities and guiding potential therapies. Raman spectroscopy therefore has the potential to provide complete non-invasive imaging through the eye. Employing this technique in a portable system would mitigate limitations and ambiguities introduced by the GCS and triage patients during the critical time before transport to emergency departments. In time-sensitive and remote settings such as in military service or at roadside or sport pitch-side settings, this would monitor ongoing health to protect patients from secondary or repeat injury and therefore long-term neurological, cognitive and psychological morbidity.TBI diagnostics is a broad field with clear objectives to improve the speed and precision of diagnostic techniques to maximise patients’ neurological recovery. However, there are few therapies mentioned when discussing medical intervention following diagnoses, another great unmet need. Advances in TBI treatments would work in synergy alongside developments in diagnostic techniques to improve the patient journey and reduce deaths from TBI worldwide. All available technologies have challenges relating to specificity and sensitivity, but the potential of RS and other techniques as new additions to the ocular and TBI diagnostics toolbox is significant and several ongoing research efforts in TBI should start to be realized in the coming years.Disclosure Statement: The authors declare no conflict of interest.