Tissue-engineered cartilage possesses obvious advantages including non-rejection, minimally invasive or non-invasive, the potential of large-scale production to ensure sufficient donors and controllable morphology. Exploration and advancements of tissue-engineered cartilaginous framework are also emerging in aspects including three-dimensional biomaterial scaffolds, acquisition of seed cells and chondrocytes, 3D printing techniques, inducing factors for chondrogenesis and so on, which has greatly promoted the research process of biomaterial substitute. This review discussed the development, current application and research progress of cartilage tissue engineering in auricular reconstruction, particularly the usage and creation of biomaterial scaffolds. The development and selection of various types of seed cells and inducing factors to stimulate chondrogenic differentiation in auricular cartilage were also highlighted. There are still confronted challenges before the clinical application becomes widely available for patients, and its long-term effect remains to be evaluated. We hope to provide guidance for future research directions of biomaterials as an alternative to autologous cartilage in ear reconstruction, and finally benefit the transformation and clinical application of cartilage tissue engineering and biomaterials in microtia treatment.Microtia is a congenital deformity of the ear with an incidence of about 0.8–4.2 per 10,000 births. Total auricular reconstruction is the preferred treatment of microtia at present, and one of the core technologies is the preparation of cartilage scaffolds. Autologous costal cartilage is recognized as the best material source for constructing scaffold platforms. However, costal cartilage harvest can lead to donor-site injuries such as pneumothorax, postoperative pain, chest wall scar and deformity. Therefore, with the need of alternative to autologous cartilage,  Microtia is a congenital ear malformation that can develop as an isolated birth abnormality or one of the manifestations of other syndromes, which can be classified based on the severity of the deformity, from mild structural changes to complete absence of the auricle . PatientMicrotia occurs in approximately 0.8–4.2 per 10,000 births . CurrentOne of the basic factors of auricular reconstruction is the supporting framework under skin to maintain the fine anatomical structure of the ear. At present, the supporting framework/scaffold mainly includes the auricular framework carved from the patient’s autologous costal cartilage, the Medpor prefabricated framework, and the tissue-engineered cartilage auricular framework. Autologous costal cartilage transplantation has been the current gold standard treatment for auricular reconstruction, which has the advantages of easy engraving, no rejection, and low incidence of cartilage exposure . In 1920Generally, the 6th, 7th, and 8th costal cartilages of the contralateral thorax were selected as the donor area for sculpting and reinforcing the main framework of the auricular framework . Howeverin vitro and in vivo study of biomaterial scaffolds, bioprinting technologies and cartilage tissue engineering have gradually become novel research hot points in auricular reconstruction research  and tissue fluid. The extracellular matrix (ECM) of cartilage is made up of collagen fibers (mostly type II collagen) supporting glycoproteins and proteoglycans with a protein core connected with glycosaminoglycan molecules such as hyaluronic acid (HA) and chondroitin sulfate , adipose-derived stem cells (ADSCs), induced pluripotent stem cells (iPSCs), articular cartilage progenitor cells (ACPCs), Chondrocytes isolated from a biopsy of patients’ autologous cartilage tissues continue to be the principal source of engineered cartilage, for its property of spontaneously secreting cartilage-specific matrix . Autologin vitro culturing of the chondrocytes in the 1960s, the optimization of isolation and amplification of chondrocytes has been extensively studied in tissue engineering  can be isolated from adipose tissue, synovium, bone marrow and umbilical cord matrix, among which bone marrow-derived MSCs (BMSCs) are especially characterized with high proliferation rate and the potential for chondrogenic differentiation under proper tissue conditions  and certin vitro and in vivo (Adipose-derived stem cells (ADSCs) can differentiate into adipocytes, osteocytes, chondrocytes, and nerve cells under different conditions. These cells have the advantages of abundant tissue sources, easily accessible sampling, low immunogenicity, rapid proliferation and multi-directional differentiation potential, which has become a research hotspot as seed cells in cartilage tissue engineering . TGF-β h in vivo . Griffinin vitro under induced directional induction and successfully generate articular matrix cartilage to repair defects after transplantation (Induced pluripotent stem cells (iPSCs) can be obtained from somatic cells through reprograming procedures, which were firstly derived from retrovirus-mediated transcription factors into fibroblasts and eventually achieve embryonic-like multilineage differentiation potential  Takahas. Indepenantation . A studyantation . Howeverantation .in vitro culture revealed that these cells possess a robust ability to proliferate without losing their multipotent differentiation capacity and to create cartilage-like matrix in the culture framework  was first identified and isolated by Gary P . from thramework , which min vitro expansion. Subsequently, after in vitro culture, they are transplanted back into the body to achieve the purpose of tissue repair or replacement , polylactic acid (PLA), and poly-caprolactone (PCL) and are widely used as medical biodegradable materials, controlled drug delivery systems and tissue engineering scaffolds . The advin vitro. The first patient presented a satisfactory reconstructed auricular morphology after the 2.5-year follow-up, and the histological examination also showed typical cartilage formation which resembles natural cartilage. Meanwhile, they also preliminarily coped with the inflammatory reaction of stent material by extending the induction time in vitro. This research provided a new strategy to improve the mechanical strength of engineered cartilage and maintain the morphology after implantation. However, the residual polymer materials are prone to cause aseptic inflammation, and the formation and distribution of chondrocytes and ECM also affect the mechanical stability, which remains the major problems to be solved in the application of synthetic material scaffolds.In 1997, During the last decade, hybrid materials consisting of natural and artificial synthetic materials have gradually become a research hotspot on account of the shortcomings of single materials. Experiments have been carried out to obtain composites with good histocompatibility, controllable morphology, and appropriate mechanical properties with the fabrication of hybrid implants. There are also studies emphasizing the combination of degradable and nondegradable materials in an attempt to help maintain the framework of the complex auricular contours.Studies have reported the usage of fibrin gel where auricular chondrocytes are suspended to cover the Medpor framework with oxidizing solution utilized to alter the implant’s surface by adding hydrophilic properties . There wDiverse biomaterial scaffold fabrication techniques have been established for processing different microstructures with controlled characteristics such as pore size, porosity, and pore interconnectivity, such that they are suited for chondrocyte growth, adhesion, and rapid nutrient transport . The mosSolvent casting and particulate leaching (SC/PL) approach requires the construction of a salt/polymer suspension that is cast using a specific solvent. Upon solidification, as a result of solvent evaporation, the salt may be dissolved out of the scaffold and exit the pores. The pore spaces involved in such scaffolds may improve the proliferation and extracellular matrix production and benefit the growth of chondrocytes. However, this SC/PL technique was reported to achieve well effect mainly in small templates . Jing etInjection modeling has been extensively studied based on natural material scaffolds such as alginate , fibrin in vitro with the same structure which is mirror-symmetrical to the normal ear. The CAD/CAM method was applied to produce a negative cast of a half-sized human ear in a mirror image. Based on this mold, it was capable to form the PGA fibers into a scaffold in the shape of a typical ear in half size. Furthermore, they enhanced the mechanical strength of the PGA scaffold by coating it with an optimized amount of PLA, which as well maintained the biocompatibility of the framework. This technique has also been actively employed subsequently in the field of auricular tissue engineering (Computer aided design and manufacturing (CAD/CAM) has been reported in the preoperative planning and the creation of patient-specific ear prostheses , and hasineering .in vitro culture. Bio-inks are cross-linked or stabilized during or immediately after printing to fabricate the desired structure  printing based on CAD/CAM enables rapid production of patient-specific anatomically attainable 3D models . Specifitructure . An ideatructure .Different natural and synthetic biomaterials with specific features have been identified as cell-laden bio-inks in different bioprinting applications . Currentin vitro immunostaining (staining . A uniquGelatin methacrylate (GelMA) is a photosensitive semi-synthetic hydrogel that when combined with a photoinitiator, it can be quickly cross-linked and solidified to produce a three-dimensional structure of particular strength. The structure contains cell adhesion sites and matrix metalloproteinase hydrolysis sites, which enables cell growth and migration . The mecin vivo  was conducted led by a regenerative medicine company  for ear reconstruction in patients with unilateral microtia (NCT04399239). The printed collagen hydrogel scaffold was made to encase the patient’s auricular chondrocytes after 3D scanning of the opposite ear which precisely matches the patient’s auricular shape. Exclusive 3D-bioprinter, bio-ink, cell culture system and implanted protective technology were designed systematically in the therapeutic production. This procedure may make breakthrough progress in the 3D bioprinting field in auricular reconstruction in the future, and the 3D-bioprinted implants may provide beneficial effects for microtia patients. Moreover, it also suggests the great significance of 3D bioprinting technique in a broader field of regenerative medicine, such as the realization of organ printing.It is estimated that approximately 100–150 million chondrogenic cells are required to generate an adult human ear-shaped cartilage , but celin vitro (Transforming growth factor β (TGF-β) family act as multifunctional components mostly produced in cartilage and bone. TGF-β1, TGF-β2 and TGF-β3 are correlated with the processes of differentiation and de-differentiation of chondrocytes as a prelude to cartilage synthesis, stimulation of type II collagen and proteoglycans as well as differentiation of MSCs . TGF-β1 in vitro . Insulinin vitro . Studiesin vitro . Combinain vitro . Fibroblin vitro . FGF-2 win vitro , while Fin vitro . Studiesin vitro .Despite the stable proliferation of auricular chondrocytes, dedifferentiation with rapid loss of chondrocyte phenotype and competence is unavoidable during repeated passages . The co-Genetic engineering technique intends to transfer cartilage growth factor genes into target cells through a carrier (such as adeno-associated virus) or attach the carrier to a biological scaffold so that the growth factor can achieve a stable and sustained expression to promote cartilage regeneration. Studies considering gene transfer of growth factors such as IGF-1, BMP-2, TGF-β and FGF-2 have been reported to lead to an enhancement in chondrogenic differentiation of MCSs . GeneticAt present, surgical procedure based on autologous costal cartilage carving and transplantation remains the major treatment of total ear reconstruction. During the past decades, a variety of research and progress has been made in the field of auricular cartilage tissue engineering. The application of 3D printing and tissue engineering in medicine holds great promise for future innovation and more consistent results for microtia patients. The 3D printing approach can shorten operation time, avoid morbidity at the donor region, produce repeatable outcomes and reduce rejection rates compared to autologous costal cartilage transplantation. However, the prohibitive cost, limited application of printers and clinical translation obstacles are still confronted challenges before these potentially revolutionary choices become available to patients.Clinical trials in human have also achieved preliminary results, but the safety and long-term effectiveness have not been fully confirmed. Before large-scale clinical application, a number of scientific and technical challenges need to be solved. The complex interrelation between cellular biochemistry, immunoreaction, and biomechanics of natural and synthetic material requires further investigation to design and fabricate the optimal individual-characterized auricular cartilage. The exploration and development of hybrid materials may be a favorable direction to obtain an ideal cartilage scaffold. The pathogenesis and molecular biomechanism of chondrogenic process in congenital microtia need to be further studied, which may contribute to the optimization of cartilage regeneration system.The research progress of tissue-engineered cartilage in ear reconstruction requires mutual promotion of multiple research fields, and further to develop a systematic and standardized model of scaffold fabrication, cell extraction and culturing, and construct implantation to realize the clinical transformation and application of tissue-engineered auricular reconstruction in microtia.


Oocyte donation (OD) has greatly improved over the last three decades, becoming a preferred practice of assisted reproductive technology (ART) for infertile women wishing for motherhood. Through OD, indeed, it has become possible to overcome the physiological limitation due to the ovarian reserve (OR) exhaustion as well as the poor gamete reliability which parallels the increasing age of women. However, despite the great scientific contribution related to the success of OD in the field of infertility, this practice seems to be associated with a higher rate of major risky events during pregnancy as recurrent miscarriage, infections and placental diseases including gestational hypertension, pre-eclampsia and post-partum hemorrhage, as well as several maternal–fetal complications due to gametes manipulation and immune system interaction. Here, we will revisit this questioned topic since a number of studies in the medical literature focus on the successful aspects of the OD procedure in terms of pregnancy rate without, however, neglecting the risks and complications potentially linked to external manipulation or heterologous implantation. Infertility is a prevalent problem in our society today. According to the World Health Organization (WHO), in the recent few decades, the number of infertile couples has greatly increased to 48 million and the number of individuals, both females and males, affected by infertility has improved to 186 million .In reality, 40 years after its disputed clinical introduction, IVF is available as a successful treatment for infertility almost all over the world. At least 2.5 million treatments are performed every year worldwide resulting in more than 500,000 births annually, while until today, over 8 million children were globally born by IVF . Such a However, a number of conditions are directly related to better results. In ART, as well as in natural pregnancies, the woman’s age still remains a pivotal factor in determining pregnancy success and the 80% success rate of IVF has been generally reported in females within the fourth decade of life . NeverthHere, we provide an attempt to revisit safety conditions and risks in gametes’ employment for ART, based on the regulatory norms and effectiveness of IVF programs in providing healthy pregnancies and newborns as reported in the medical literature.The most recent annual reports of the European IVF-Monitoring (EIM) consortium, in agreement with the European Society of Human Reproduction and Embryology (ESHRE), have recently collected data from 39 European countries describing treatments employing donor gametes (DG), which have reached up to 8% of all ART cycles .In the last decade, Spain confirmed its role as the first European country performing ART with DG with 37,618 egg donation (ED) cycles yearly, followed by Russia with 9804 cycles [In 2018, European ED number estimation included 36,938 donations and 24,148 of them were employed for fresh embryo transfer (ET), whereas 16,130 procedures included frozen oocytes. However, these data are underestimated since several country performances are missing. Overall 25,760 deliveries were reported with ED. Furthermore, pregnancy rate per ET is higher by freshly donated (49.6%) than for thawed oocytes (44.9%), thus preferring fresh oocytes as the best choice when using this technique [IVF treatment success with patient’s egg employment decreases progressively with the woman’s age in relation not only for the low number but also for the reduced availability of healthy oocytes. Since just 25 countries set age limitations for egg donors generally between 18 and 35 years . ESHRE dMoreover, deliveries are also increased between 2014 and 2018 and this is primarily due to the higher contribution of Spain with respect to other countries ,7. On thRecurrent miscarriages (RMs) are a critical issue in public health, often responsible of psychological troubles in women’s health and inevitably within the couple relationship. Beyond chromosomal aneuploidy ,11, seveHowever, despite the IVF programs that allow one to perform good-quality ET by pre-implantation genetic testing for aneuploidy (PGT-A), many infertile couples undergo recurrent implantation failure with RMs and fail in having pregnancies . A systeIntravenous immunoglobulin (IVIg) is an established treatment for many autoimmune and inflammatory diseases . DespiteConception is not the only concern for infertile couples with a desire of parenthood. Although, a successful ART cycle is not flawless and is associated with an increased risk of adverse maternal and perinatal outcomes as compared to spontaneous gestation, the procedure has been reported as associated with increased risk of gestational hypertension, pre-eclampsia, diabetes mellitus, intrahepatic cholestasis, placenta previa, placental abruption, preterm rupture of membranes, placental adherence, postpartum hemorrhage, polyhydramnios, preterm labor, low birth weight, and small-for-date infant .It is conceivable that a large part of pregnancy complications associated with fertility treatment is due to multiple gestations that are more frequent in ART conceptions. However, the high risk of placental complications after fertility treatment appears to be independent of multiple gestations . IVF hasAlthough some meta-analyses have shown that ART is associated with an increased risk of pre-eclampsia, as reported in The types of ART protocols also appear to play a role in differences in maternal and perinatal outcomes . In factGestational diabetes has also been studied as an outcome in IVF pregnancies because it predisposes to a higher risk of pregnancy-related hypertension, fetal macrosomia, operative delivery, and cesarean delivery . An incrAlso, an increased risk of preterm delivery appears to recur in pregnancies conceived through IVF . ParticuAbnormalities in birth weight, namely as low as less than 2500 g, have been associated with IVF . The assSeveral studies highlighted a concern for potentially increased risks of primary defects in ART pregnancies. Hansen and coworkers in their meta-analysis showed a moderate increase in the risk of birth defects in ART-conceived children compared with naturally conceived ones . AnotherOther interesting topics are imprinting disorders and neurodevelopmental disorders in IVF pregnancies, but data are still lacking and questionable ,53.Finally, there is evidence that at least some IVF-conceived children may be at increased risk for cardiometabolic disorders including insulin resistance, higher blood pressure and higher body fat percentage compared to children conceived without medical assistance. Most IVF children are still in their first three decades of life and, thus, limited data exist regarding long-term morbidity and mortality .Epigenetic derangements are crucial in gametogenesis. In particular, DNA methylation is involved in different stages of oocyte maturation and seems to be strongly influenced by hormonal stimulations mainly required for multiple follicular growth induction related to ART. Thus, ART procedures are apparently associated with increased imprinting and DNA methylation errors . ExploriThe DNA of gametes, is sensitive to the effects of oxidative stress through methylation of nucleotide bases, specifically in the CpG islands. Cytosine and guanine suffer the higher effects, with the latter being the most susceptible. The products of their oxidation are 8-oxo deoxyguanosine and 5-hydroxyme-thylcytosine . MoreoveFurthermore, controlled ovarian stimulation required in ED cycles also increases the homocysteine levels with an associated inhibition of oxidative stress and methylation in follicular fluid, resulting in an inducing in epigenetic derangements in the zygote ,61. To s+ T lymphocytes were also demonstrated as compared to spontaneous or IVF pregnancies, thus indicating a different stimulation of intracellular cytokines [Pregnancies from ED generate an immunological paradox since the fetus is allogeneic to the mother and hence exposes non-self antigens and cells . Neverthytokines . Moreoveytokines .Alterations in the immunological pattern also impacts on placenta morphology in OD pregnancies. Notably, villitis of unknown etiology and chronic deciduitis have both been found to be associated with chronic inflammation . From a In women with pre-eclampsia in naturally conceived pregnancies, the thrombomodulin serum levels degradation products are higher than in uncomplicated naturally conceived pregnancies . ThromboEven if pregnancy is a physiological condition, trophoblasts and decidua play an important role in maternal–fetal tolerance, in order to avoid possible risk related to any immunological rejection. After ranging decidua, trophoblasts differentiate into villi (without HLA molecules on their surface) and extravilli (EVT) which express polymorphic HLA-C both of maternal and paternal origin, and non-polymorphic HLA-E and HLA-G, responsible for non-specific immune response.HLA-C molecules act as ligand for killer immunoglobulin-like receptors (KIRs) on decidual NK cells and the different combination between KIR aplotypes and HLA-C allotypes is responsible for immune tolerance against fetus or increased risk of pregnancy complication .+ cells, suppressing their cytotoxic activity and establishing allorecognition [+ cells through Major Histocompatibility Complex-II (MHC-II) molecules: this interaction represents the so-called indirect antigen recognition between mother and fetus  response is crucial for successful implantation  and to p+ macrophages, CD4+ T cells and Treg cells, while in OD ones, either complicated or uncomplicated, this condition occurs independently from the occurrence of pre-eclampsia. Moreover, inflammatory lesions and maternal M2 macrophages found in the chorionic plate were linked to 0% of pre-eclampsia incidence in OD pregnancies, while a high incidence of pre-eclampsia is apparently associated with the absence of inflammatory signs, thus hypothesizing that the potential inflammatory state could perhaps avoid the fetus rejection [In contrast with natural pregnancies, ED implies the mother’s immune system overexposure to several non-self HLA-C antigens  that seem to induce higher PE incidence, but also a decrease in live birth rate, especially after double ET ,82. Uncoejection .Since the reliable immune system function is related to good pregnancy outcomes even in the case of donor gametes, it is interesting to investigate how it could react to an embryo graft in pregnant women affected by autoimmune diseases (ADs). Even if the literature is still lacking in understanding the connection between immune system and ART procedures in ADs, recently, Simopoulou and colleagues  tried toA deeper knowledge of the AA roles in pregnancy is thus needed which depicts a new frontier for pregnancy-related risk prevention and additional clinical studies are also required to better investigate this fascinating field of research in both natural and ART-driven pregnancies.The corpus luteum (CL) is a transitional organ representing the main source of steroid hormones, and vasoactive and angiogenic substances, which play a critical role in the early phase of pregnancy . Embryo CL produces several metabolites with angiogenic or anti-angiogenic effects. In the early and late luteal phase, 16-ketoestradiol (16-ketoE2) and 4-hydroxyoestrone (4-OHE1) are, respectively, produced with a pro-angiogenic effect by stimulating VEGF production . Since hInfertility is both a medical and social condition that affects a large female population worldwide and is certainly a disease. It is related to several pathophysiological circumstances and its pathogenesis is often unclear, with relative indecision to establish the appropriate treatment choices. In the last three decades, many treatments have been introduced for this disease and the field of reproductive medicine is currently in rapid evolution for the employment of novel therapeutic approaches, primarily based on IVF techniques.However, despite the fact that IVF treatments with own or donor gametes significantly increase the couple’s chances of becoming parents, they also carry several maternal and neonatal risks principally related in procedures using heterologous gametes. In fact, this practice seems to be associated with a higher rate of major risky events during pregnancy as recurrent miscarriage, infections and placental diseases such as gestational hypertension, pre-eclampsia and post-partum hemorrhage, as well as to several maternal–fetal complications due to the immune system interaction with the effects of the in vitro manipulation of gametes. Furthermore, both processing and culturing techniques of oocytes as both autologous or heterologous gametes, could result in altered gene methylation responsible of epigenetic derangements. Hence, in the era of precision medicine, to overcome these limitations, it would be desirable for the community of women undergoing IVF as well as for physicians, to test such a potential risk by exploring the integrity of gene methylation expression in both own or donated eggs.In conclusion, the identification of several potential risks in ART procedures related to OD is essential in preventing possible fetal–maternal diseases and necessary for clinicians to address proper screening and surveillance during IVF related pregnancies.


As the leading cause of cancer-related mortality, lung cancer continues to pose a menacing threat to human health worldwide. Lung cancer treatment options primarily rely on chemoradiotherapy, surgery, targeted therapy, or immunotherapy. Despite significant progress in research and treatment, the 5-year survival rate for lung cancer patients is only 10–20%. There is an urgent need to develop more reliable preclinical models and valid therapeutic approaches. Patient-derived organoids with highly reduced tumour heterogeneity have emerged as a promising model for high-throughput drug screening to guide treatment of lung cancer patients. Organoid technology offers a novel platform for disease modelling, biobanking and drug development. The expected benefit of organoids is for cancer patients as the subsequent precision medicine technology. Over the past few years, numerous basic and clinical studies have been conducted on lung cancer organoids, highlighting the significant contributions of this technique. This review comprehensively examines the current state-of-the-art technologies and applications relevant to the formation of lung cancer organoids, as well as the potential of organoids in precision medicine and drug testing.Video AbstractThe online version contains supplementary material available at 10.1186/s12964-023-01332-9. Lung cancer is currently the most common type of cancer worldwide, with the highest mortality rate compared to other forms of cancer. It accounts for approximately 11.4% of all cancer cases and 18.0% of cancer-related deaths worldwide .In recent years, notable progress has been made in the field of targeted anticancer drugs and immunotherapy for lung cancer, but challenges regarding drug efficacy, toxicity and drug resistance remain. For example, lung adenocarcinoma exhibits marked inter- and intratumoral heterogeneity, which may lead to treatment failure and resistance development . TherefoOrganoids are in vitro 3D mini "organs" that highly mimic the pathophysiological system of the human body and can be created from embryonic stem cells (ESCs) , spermatTo date, biobanks for lung cancer organoids have been established from multiple subtypes of lung cancer tissues of primary biopsy or surgery, including adenocarcinoma , squamouRepresentative of traditional two-dimensional culture, cell lines remain the most common disease models. While cell lines have the benefits of low cost, high throughput, and simple culture conditions, their disadvantages have become increasingly apparent. One important property of cell lines is the unavoidable genetic drift that occurs over time, resulting in progressive accumulation of genetic mutations. This will cause significant damage to further studies if the phenotypic and genetic heterogeneity in tumour cells is altered . AdditioPDX models are frequently generated using immunodeficient or humanized mice, which are transplanted in situ or ectopically once the tumour load reaches a certain level. Compared with cell lines, PDX models better preserve key features and interactions between the tumour and its stroma . In addiThe ability of organoids to self-renew and self-proliferate while retaining the specific functions and structures of native tissues has generated significant interest in their use for disease modelling . The speThe core principle that governs development of diverse normal tissues and tumour organoids remains largely uniform, primarily hinging on creation of a three-dimensional microenvironment through matrix adhesives. Various nutrients that maintain cell stemness are added to the culture system to promote cell growth and development. Following dissociation of cancer tissues, several procedures, including physical cutting, chemical digestion, and filtration, are necessary to degrade a suitable lung cancer sample into unbound cells and cell clusters. Then they are suspended in a liquid extracellular matrix (ECM). The mixed matrix gel is inoculated on plates and allowed to solidify Fig. . AfterwaEstablishment of organoids has the following priorities: ① find and obtain tissue/organ stem cells; ② create cultivation environment; and ③ maintain cell stemness. Most of the cells used for organoid culture are derived from stem cells including tumour stem cells derived from the desired tissue or organ because of their strong cell stemness. For example, the addition of low concentrations of PF-127 to the cell culture medium has been found to promote the differentiation of human embryonic stem cells (hESCs) into organoids . In 2021Traditional organoid culture systems have limitations in terms of variability and scalability. Additionally, existing technologies struggle to achieve large-scale production of homogeneous organoids for drug screening. To improve the accuracy and stability of organoid research, researchers have been studying microfluidic devices with microfluidic chips as the core. These chips are used to construct and simulate the human tissue microenvironment, creating a microphysiological system that closely resembles the human body. Through dynamic flow in microfluidic devices for waste exchange, oxygen and nutrient supply, organoids can be precisely managed and uniformly cultured in large numbers while preserving a complex microenvironment . The devTo culture organoids, hydrogels must provide the necessary physical factors such as space and pressure stimulation, as well as the nutrients required for organoid growth. This creates an environment that mimics the natural physiological conditions in which stem cells are found in vivo, and provides cancer cells with an extracellular component that is rich in nutrients and can stably support 3D structures. The hydrogel acts as a scaffold for stem cells to adhere to and self-assemble into specific organoids. Because this hydrogel is man-made, it is easy to control its chemical composition and critical properties and ensure consistency without posing a risk of infection or immune response; it is therefore significantly better than traditional Matrigel. Wilkinson et al. developed a self-assembling lung organoid platform  with var3D bioprinting is a tissue engineering method that uses software to rapidly manufacture biomedical products by layering and controlling the positioning and assembly of biomaterials. This technology has the potential to replace artificial methods and construct complex heterotypic tissues, improve automation and biomimicry in model quality, and provide highly consistent, reproducible, and scalable organoids. As a result, bioprinting technology is showing great potential in high-throughput screening and individualized precision treatment. In their study, Melissa and colleagues were able to culture renal organoids using 3D bioprinting technology achieving morphology, component cell types, and gene expression levels comparable to those constructed manually . This teOrganoid technology has been acknowledged as a promising biotechnology in precision medicine. Vlachogiannis and colleagues conducted a pioneering study on organoid drug sensitivity testing and in vivo consistency in gastrointestinal tumours, yielding remarkable results . SubsequIn the past, the development and screening of many antineoplastic agents was performed using standard cell lines, but they proved ineffective in clinical studies . HoweverBiobanks have been established due to their advantages of low cost and easy maintenance organoids-based. However, effective diagnosis and treatment programs for glioblastoma (GBM) tumours are challenging due to their high inter- and intratumoural heterogeneity. To address this issue, Jacob established glioblastoma organoid (GBO) biobanks, which retain the key characteristics of GBM. This resourceful biobank provides a rich source for inositol research and translational research in GBM, further reflecting the significant advantages of GBO in testing personalized treatment . The estOrganoid models have been established for various tumours and diseases, including PDOs applied to primary cultures of lung cancer cells. These models serve as ex vivo models to accurately reproduce clinical responses to chemotherapy and immunotherapy in precision treatment . Zhang cCRISPR‒Cas9 is an adaptive immune defence that evolved over time in prokaryotes to combat invading viruses and exogenous DNA. CRISPR‒Cas9 technology enables specific DNA modification of targeted genes, making it a valuable tool in gene screening and disease treatment. Hai and colleagues used CRISPR‒Cas9 to knock out multiple oncogenes from Cre-dependent SOX2 knock-in mouse lung organs and created a lung squamous cell carcinoma model that resembles human disease at genomic and phenotypical levels . PD-1 inHigh-throughput single-cell sequencing technology has become a crucial tool in exploring biological questions such as tumour heterogeneity through analysis of the genetic material of individual cells. This method allows for identification of heterogeneous cells through cell fractionation and study of interactions between tumour cells . For exaNa et al. constructed a primary, in situ, driver gene defined mouse model of SCLC using organoid culture, gene editing and orthotopic transplantation. The model is expected to be applied to basic and translational studies of SCLC. In addition, the team found that loss of the epigenetic gene KMT2C promotes SCLC metastasis and malignant progression. It is anticipated that SAM can prolong survival periods and improve quality of life for patients with KMT2C mutations, providing a new therapeutic target for SCLC patients . Lung sqApplication of patient-derived organoids is now prevalent in basic and translational studies of lung cancer. However, certain restrictions currently limit the potential applications of organoids in personalized medicine and drug development. The absence of a vascular system, a distinct immune microenvironment, and variations in cell types and extracellular matrix from in vivo tissues and organs present obstacles for organoids with regard to replicating the authentic microenvironment and drug reaction.Currently, most organoids lack in vitro vasculature, leading to necrosis in exploding tumours. Nevertheless, the 3D bioprinting technique has effectively generated centimetre-scale intestine-like tissues that are highly biomimetic, comprising tubular structures, branched vessels and small intestinal epithelial-like crypts . A studyConstructing tumour organoids that accurately mimic interactions between tumour cells and extracellular matrix is a significant challenge due to the complex cellular components present in the TME. Macrophages, which are abundant in the tumour immune microenvironment, have been found to promote cancer cell invasion through the COX-2/PGE2/β-catenin signalling pathway by inducing the epithelial-mesenchymal transition (EMT). Coculture experiments with human lung cancer cells exhibit this phenomenon . ConsequThe extracellular matrix (ECM) is composed of several proteins including collagen, laminin, and cell cohesion . Due to Dijkstra et al. combined genetic and histomorphometric analyses, and observed overgrowth of normal airway organoids in most lung cancer organoids derived from over 70 NSCLC samples . The ratOrganoids have emerged as one of the most promising biomedical technologies because of their high correlations with in vitro characteristics and drug response. Additional research into these models has resulted in emergence of organoid technology, which is now used for disease modelling, personalized medicine, drug development and screening. The high correlation between organoid-based drug response and clinical outcome has been revealed by real-world studies and clinical trials , 93. ConTo establish reliable organoid models, researchers should focus on combining organoids with advanced technologies and ensuring their validity in real-world environments. Forthcoming research on organoids in lung cancer will concentrate on various aspects. First, establishment of standards for organoid culture, quality control, and drug testing is still necessary. Automatic approaches for organoid culture and drug tests will be investigated, such as microfluidic chips and automated workstations, to reduce the instability due to manual operations. Moreover, adoption of a complex model that encompasses the vascular system, immune cells, fibroblasts, and other components to simulate the tumour microenvironment is indispensable for advancing the biological significance and accuracy of in vitro tests. As the culture techniques of LCOs progressively develop, this model will indubitably assume greater significance in personalized medicine, drug development and basic research of lung cancer.


This oxidase is also a key mitochondrial enzyme for cellular bioenergetics, especially for the nerve cells in the retina and brain. In addition, dopamine (DA)-enhanced release of nitric oxide may also be involved in controlling myopia by activation of nitric oxide synthase, enhancing cGMP signaling. Recent evidence has also suggested that RLRL may inhibit myopia progression by inhibiting spherical equivalent refraction (SER) progression and axial elongation without adverse effects.Repeated low-level red-light (RLRL), characterized by increased energy supply and cellular metabolism, thus enhancing metabolic repair processes, has gained persistent worldwide attention in recent years as a new novel scientific approach for therapeutic application in myopia. This therapeutic revolution led by RLRL therapy is due to significant advances in bioenergetics and photobiology, for instance, enormous progresses in photobiomodulation regulated by cytochrome In this review, we provide scientific evidence for RLRL therapy as a unique paradigm to control myopia and support the theory that targeting neuronal energy metabolism may constitute a major target for the neurotherapeutics of myopia, with emphasis on its molecular, cellular, and nervous tissue levels, and the potential benefits of RLRL therapy for myopia. Myopia is caused by the parallel light penetrating via the eye refractive system and focusing the front retina as the eye is relaxed, which causes blurred images Myopia has increasingly become a major public health problem, causing visual damage worldwide Myopia is linked closely with many complications, for instance, cataracts, macular degeneration and vision loss Previously, various interventions, including atropine, orthokeratology, spectacle and soft contact lenses, defocus incorporated multiple segment (DIMS) lenses and their combinations have been applied to slow down myopia progression with limited success Light is defined as a form of electromagnetic radiation manifested by particles and wave properties. The waves of electromagnetic radiation possess unidirectional vectors, specified by wavelengths , frequencies (oscillations per second), and amplitudes (difference of trough and peak). The energy particles within electromagnetic radiation include photons, which travel at 3×108 m per second. Thus, mixture of various waves would elicit photons moving at different amplitudes and frequencies that are scattered and absorbed, reflected by various objects, for instance, biological materials.By contrast, red-light is monochromatic, allowing high specificity as to molecular biomodulation. Red-light can be generated by lasers or light-emitting diodes (LEDs), appliable to photobiomodulation (PBM) in eyes. Lasers may generate coherent light energy efficiently to allow penetration to various tissues. The beam width of the lasers can be strengthened by coupling with fiber optic to permit energy distribution to larger areas. LEDs may generate efficient light without coherence in wavelengths of 4-10 nm. However, LEDs generate negligible heat, without any thermal injuries More recently, effects of different light wavelengths rather than those of different intensities on myopia onset and progression have drawn increasing attraction. Light waves are electromagnetic with different frequencies, including infrared rays, ultraviolet rays, radio waves, visible light , referring to that induced myopia may be achieved by deprivation of form vision from eyes in a status of susceptibility, for example, lid-suture 2) or high (0.10 J/cm2) energy densities respectively c oxidase (CCO), resulting in better energy metabolism within mitochondria Red light may promote biological processes at a low dose but inhibit biological process at a high dose, in a U-shape response. Hormetic curves are better to predict biological responses within pharmacological threshold than linear curves c oxidase, a primary light photoreceptor in the red and the near-infrared electromagnetic spectrum Although many photoreceptors have been discovered, the mechanism of low-level red-light remains unclear since many molecules have photoreceptor capabilities. For instance, reactions from a molecular target induced by a wavelength can be complex. For example, NADH-dehydrogenase and flavoprotein have various photoreceptors from the violet to blue or from red to near-infrared spectral areas Among the possible hypotheses for relationship of myopia and light intensity, the most possible hypothesis is that the light promotes synthesis and release of dopamine (DA) by retina DA may enhance synthesis and release of NO (nitric oxide) to mediate eye growth via activation of NOS (nitric oxide synthase). NO may decrease form deprived myopia dose dependently while NOS inhibitors may block myopia inhibition regulated by atropine Red light may induce oxidation in cytochrome c via activating CCO to increase use of oxygen, to elevate mitochondrial membrane sensitivity and pore permeability With treatment of monochromatic light, the symmetric myopic sign-dependency is manifested, resulting in the spherical defocus on the ocular acuity. The observed results suggest that the human visual system may integrate the chromatic differences in refraction to discern the signs of defocus PBM may also activate TGFβ/Smad signaling in wound healing to mediate myopia progression PBM or low-level light therapy, via photon irradiation, has decades of clinical applications to soft tissue injuries due to promoting wound healing. Low-level red-light can penetrate tissues such as the brain, heart and spinal cord PBMT (photobiomodulation therapy) has increasingly become popular in human medical therapies recently. Recent reports have suggested that laser therapy can be used for treatment of inflammation and promotion of wound healing Myopia pandemic affects a large number of population in the world Recently, several reports have indicated that RLRL may inhibit myopia progression by inhibiting SER progression and AL elongation Effects of various lights on myopia may be quite different. For instance, red-light, a long wavelength light should lead to hyperopia defocus Myopia has been increasing steadily, especially in East Asia. Red-light may improve choroidal blood perfusion and may be a critical tool for control of myopia via cytochrome and nitric oxide signaling. In all, application of red-light may retard myopia effectively, possessing high potential for prevention of myopia onset and control of myopia progression. However, the safety and the implementation of RLRL therapy are two biggest issues for extensive investigations. In addition, whether such a therapy is effective in high myopia requires further scientific studies. Furthermore, whether RLRL therapy can prevent onset of myopia remains unknown. Finally, whether RLRL therapy can be used to replace atropine therapy for myopia control needs comparative studies.


Because, in many previous studies, the factthat Pregnancy-Specific Beta-1-Glycoprotein is closely associated with some adversepregnancy outcomes such as small-for-gestational age fetuses (SGA), spontaneous pretermdelivery and preeclampsia supports our prediction (However, the fact that some clinical and laboratory parameters were not evaluated betweengroups may have affected the results of the study. For example, basic hematologicalparameters such as fasting plasma glucose and lipid profile are not mentioned in themethodology. However, it has been shown that fasting blood glucose and maternal lipidprofile are associated with fetal growth retardation in the absence of chronic disease(In addition, are clinical features specific to pregnancy that may be associated withfetal growth retardation, such as gestational hypertension, gestational diabetes, andpreeclampsia, have been recorded (Finally, Pregnancy-specific β-glycoprotein 1 is synthesized by the human placentaand secreted into the maternal circulation in an increasing concentration fromtrophoblast differentiation to term (We believe that the evaluation of these parameters in the study will further increase theclinical importance of the study.


High-level football (soccer) players face intense physical demands that result in acute and residual fatigue, impairing their physical performance in subsequent matches. Further, top-class players are frequently exposed to match-congested periods where sufficient recovery times are not achievable. To evaluate training and recovery strategies, the monitoring of players’ recovery profiles is crucial. Along with performance and neuro-mechanical impairments, match-induced fatigue causes metabolic disturbances denoted by changes in chemical analytes that can be quantified in different body fluids such as blood, saliva, and urine, thus acting as biomarkers. The monitoring of these molecules might supplement performance, neuromuscular and cognitive measurements to guide coaches and trainers during the recovery period. The present narrative review aims to comprehensively review the scientific literature on biomarkers of post-match recovery in semi-professional and professional football players as well as provide an outlook on the role that metabolomic studies might play in this field of research. Overall, no single gold-standard biomarker of match-induced fatigue exists, and a range of metabolites are available to assess different aspects of post-match recovery. The use of biomarker panels might be suitable to simultaneously monitoring these broad physiological processes, yet further research on fluctuations of different analytes throughout post-match recovery is warranted. Although important efforts have been made to address the high interindividual heterogeneity of available markers, limitations inherent to these markers might compromise the information they provide to guide recovery protocols. Further research on metabolomics might benefit from evaluating the long-term recovery period from a high-level football match to shed light upon new biomarkers of post-match recovery. Overalltch-play . The phytch-play , and platch-play . Howevertch-play . Adding tch-play . Adequattch-play . To ensutch-play .Fatigue is a multifactorial concept that involves both central and peripheral mechanisms . Muscle Exercise-induced muscle fatigue is a complex phenomenon that involves multiple underlying mechanisms . BrieflyStudying circulating levels of biochemical markers can shed light on the underlying mechanisms involved in post-match recovery. However, as no single biomarker can fully capture the player’s physiological response to football match-play, researchers have explored a range of analytes. Furthermore, there is a need for identifying new biomarkers that overcome limitations of those currently available . Novel e®. Peer-reviewed English-written studies published from inception to December 2022 were screened to identify eligible articles. Relevant search terms  in combination with Boolean operators  were used, and references from retrieved articles and previous reviews were screened to further identify other records. Inclusion criteria consisted of studies 1) enrolling male or female semi-professional or professional football players; 2) using official or friendly football matches as exercise protocol, or validated match simulation protocols; 3) reporting data on circulating biochemical markers to evaluate players’ post-match physiological response and/or recovery. Studies conducted in amateur players, and enrolling players with mean age < 16 years were excluded, yet some of these studies were cited and specified when necessary. To help visualize the information, articles reporting fluctuations of these biomarkers after single matches or several matches separated by at last 3 days of recovery time are presented in To provide a comprehensive overview of the current state of the art and discuss recent advancements and future perspectives on the use of biomarkers of football post-match recovery, a non-systematic approach was adopted. An exhaustive search of the scientific databases and search engines ScienceDirect, Web of Science, and Google Scholar was conducted by one reviewer. PubMed (Medline) and EMBASE were also screened using the search tool ProQuest DialogFootball training sessions and competitive matches expose the players to EIMD produced mainly as a consequence of the high eccentric muscle contraction demands that characterize common football-related movements  . FurtherSerum creatine kinase (CK) is the most commonly measured analyte in the assessment of EIMD as greatest increases in serum CK activity are produced in response to intensive eccentric compared to concentric contractions . CK is aRegarding football, a vast number of studies conducted in male and female semi-professional and professional football players have reported values of CK as a means to assess EIMD in training and competition . Of noteBesides high-intensity training periods, important increases in circulating CK levels immediately after a match or a simulated football match test are typically regarded as a sign of EIMD produced during match-play . SeveralRegarding periods of match-congestion, higher CK levels are typically observed along with increasing number of matches played. On this subject, Several efforts have been made to address the high inter-individual variability in CK data. Bayesian statistics utilizes prior knowledge in the form of prior distributions and combines it with observational data to determine the posterior distribution, thereby predicting future outcomes . Using a+ . LDH constitutes a group of oxidoreductase isoenzymes  present in almost all body tissues, which plays an important role in energy metabolism as it catalyzes the reversible conversion of pyruvate to lactate with the concomitant conversion of NADH to NAD+ . Paralle+ . Compare+ . In high+ , with ci+ , and rem+ . Nonethe+ . In addi+ , who mig+ . Like CK+ . The lac+ .Besides CK and LDH, some studies have reported circulating levels of aspartate transaminase (AST) and alanine transaminase (ALT) [formerly known as oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT)] to assess EIMD in male and female elite football players . AlthougBased on these studies, supporting the assessment of functional and perceptual fatigue measurements with individualized ranges of CK, and/or combinations of different EIMD biomarkers such as LDH might overcome limitations inherent to individual markers and provide useful insights into the monitoring of post-match recovery.A key consequence of the structural disarrangements of the skeletal muscle membranes produced during the early stages of EIMD consists of the accumulation of calcium ions. This subsequently triggers an inflammatory response proposed to be involved in different aspects of muscle repairing and remodeling processes . The timAs part of the acute-phase of the inflammatory process, cytokines are produced by leucocytes and the muscle itself (the latter being termed “myokines”) , some ofAs IL-6 levels might reflect the need for glycogen replenishment and might further increase following EIMD, authors have resourced to the analysis of this cytokine to characterize the inflammatory response produced during the post-match recovery process . Among tIL-6 is produced in larger amounts compared to any other cytokine in response to high intensity exercise. The initial release of IL-6 usually precedes a less pronounced increase in the levels of different pro- and anti-inflammatory cytokines, with IL-1β and tumor necrosis factor α (TNF-α) being the most widely studied . In partC-reactive protein (CRP) is a protein synthetized by the hepatocytes, whose circulating levels are markedly impacted by acute inflammatory stimuli . In thisIn summary, the postgame response of cytokines such as IL-6 seems to reflect adaptative changes and the muscle’s attempt to restore homeostasis following intensive exercise while increments in CRP levels may characterize the secondary inflammatory process as a consequence of muscle damage induced by match-play. While changes in IL-6 levels seem to be rather acute, monitoring post-match CRP levels might assist couches and health professionals in the monitoring of match-induced inflammatory process during post-match recovery, and can supplement the information provided by other markers.Both male and female athletes competing in highly aerobic sports typically display substantially lower white blood cells (WBC) counts compared with standard clinical reference values at rest . HoweverThese changes in WBC dynamics in response to exercise have been proposed to be consequence of different factors including cytokines release  and secrTotal WBC counts have been consistently reported to increase ≈20–100% within the first hour after a football match or match simulation in male and female elite players . ChangesMatch-induced perturbations in total WBC counts are usually restored within 24–48 h of recovery time . NonetheBesides white cells, immunoglobulins have received great attention as markers of immune status in sports. Among them, immunoglobulin A (IgA) is the most abundant immunoglobulin in external secretions thus constituting the first line of defense against invading pathogens . IgA assAs reviewed by Physical exercise constitutes a stressing situation that threatens to alter body homeostasis . ConsequEarly increases in testosterone levels following exercise are proposed to be caused by decreased plasma clearance  while suAthletes typically display peak serum and salivary cortisol levels minutes after high-intensity exercise . ExtrapoChanges in cortisol levels might reflect the psychophysiological stress induced by match-congested periods, which in turn might impair physical performance in subsequent matches . Mohr etResearch on hormonal changes in response to elite female football matches is scarce and differences owing to menstrual cycle phases and the lower physiological levels of testosterone in women are to be expected. On this subject, immediate increases in serum free testosterone levels in response to a football match were reported by Overall, current evidence suggests that football matches are capable of inducing immediate endocrine responses in professional and semi-professional male football players as denoted by increased cortisol and decreased testosterone levels. However, most of the studies conducted to date have evaluated acute changes in hormones levels in response to a football match while evidence on long-term effects is inconsistent and seems insufficient to recommend the monitoring of hormones during the post-match recovery period.Free radicals are molecules or fragments of molecules possessing an unpaired electron . Among tPostulated mechanisms by which free radicals might reduce muscle force production are multiple and englobe impairment of membrane excitability through inhibition of the sodium-potassium-ATPase-pump , inhibitNumerous molecules have been explored as biomarkers of oxidative stress with different degrees of evidence, being typically categorized into primary oxidants, oxidated molecules, antioxidative molecules, and antioxidative enzymes . In the in vivo thus contributing substantially to plasma total antioxidant capacity , a tripeptide which is oxidized to glutathione disulfide (GSSG) in the presence of free radicals in a reaction catalyzed by GPX. Glutathione reductase can then catalyze the regeneration of GSH from GSSG using NADPH as an electron donor . Thus, GChanges in the total activity of antioxidative enzymes such as CAT, SOD, and GPX have been explored to assess oxidative stress in response to exercise . CAT actThe blood levels of lipid peroxidation by-products reflect oxidative damaged inflicted to muscle cell membranes probably as a consequence of the production of free radicals by phagocytic cells . The degBased on the above-mentioned studies, football matches might cause a pro-oxidative insult in semi-professional and professional football players denoted by increased levels of oxidated molecules and upregulation of antioxidative systems. If this pro-oxidative status can accumulate throughout subsequent matches is not obvious as studies evaluating the impact of congestive match schedules on different oxidative markers have yielded inconsistent results . FinallyAs commented, serum creatine kinase is the most widely used biomarker of recovery in competitive sports such as football as it remains elevated several days into post-match recovery denoting the occurrence of EIMD . HoweverIn addition to commented limitations, some practical aspects related to the monitoring of these biomarkers should be noted. The assessment of most available biomarkers relies on the analysis of venous or capillary blood samples through laboratory tests or point-of-care platforms . AlthougIn the last decades, there has been a dearth of research pertaining to the evaluation of novel biomarkers of recovery from exercise, and the analysis of available biomarkers has been suggested to only provide a narrow picture of the complex changes that the human body undergoes during exercise . The appSportomics is a holistic approach which encompasses the simultaneous analysis of numerous metabolites through the use of analytic techniques, such as gas and liquid chromatography, mass spectrometry, and nuclear magnetic resonance in biological samples in combination with data processing methods to investigate the metabolic changes induced by exercise . These aOne of the first studies focused on football match-induced fatigue was conducted by On this subject, Long time-course analyses in sportomics studies are needed but face various challenges. Future studies should carefully monitor dietary intake and training sessions during post-match recovery to ensure the ecological validity of observed data . AdherenIn the same line, sex-based differences in match loads and activity patterns have been reported, yet few studies have explored time-course differences in circulating biomarkers during post-match recovery in elite female players, in alignment with a recent meta-analysis . On thisFootball match-play puts a physical strain on high-level football players denoted by a decline in physical performance that can persist over days. The monitoring of biomarkers provides an objective measurement of the players’ internal load during post-match recovery, which might supplement the assessment of functional and subjective indicators. However, no single biomarker can delineate the complex processes that comprise exercise recovery and aspects such as timing, sample type, technical limitations, inter-individual variance, and contextual factors make challenging the extrapolation of data reported in single studies. The use of biomarkers panels, and the individualization of physiological ranges might provide a more robust assessment of players’ fatigue during recovery, yet further research on fluctuations of different analytes throughout post-match recovery is warranted. The application of metabolomics to football research might support the identification of novel biomarkers and might also provide information on individual’s metabolomic profile to guide personalized recovery protocols. Nonetheless, most metabolomics studies conducted to date have focused on acute fatigue processes thus neglecting the long-term impact of football competition. Future studies might benefit from differentiating the metabolic disturbances that characterize the acute and residual post-match fatigue response.


Breast cancer care has seen tremendous advancements in recent years through various innovations to improve early detection, diagnosis, treatment, and survival. These innovations include advancements in imaging techniques, minimally invasive surgical techniques, targeted therapies and personalized medicine, radiation therapy, and multidisciplinary care. It is essential to recognize that challenges and limitations exist while significant advancements in breast cancer care exist. Continued research, advocacy, and efforts to address these challenges are necessary to make these innovations accessible to all patients while carefully considering and managing the ethical, social, and practical implications. Breast cancer is the commonest cancer among women and ranks as the second cause of cancer-related deaths in women . Breast While there have been significant advancements in breast cancer care, several ethical, social, and practical challenges have arisen along with these innovations.In this review, we aim to describe the innovations as well as future prospects in breast cancer care, along with their limitations and challenges.Peer-reviewed published scientific literature and continuing scientific studies were searched using relevant MeSH terms in the MEDLINE database for “innovations” OR “biomedical technology” AND “breast cancer” OR “breast surgery”. All levels of evidence were considered. Only articles published in the English language were reviewed.We report innovations in breast cancer care, possible future applications based on the published literature and continuing scientific studies, and its potential limitations and ethical considerations.The standard modality for screening and diagnosing breast malignancy has been mammography since its use following the Forrest Report in 1986, which concluded that mammographic breast cancer screening was effective in reducing breast cancer mortality and recommended the establishment of a national breast cancer screening program in the UK for women aged 50 to 64 years [In addition, contrast-enhanced mammography (CEM) is a newly emerging tool in breast radiology which uses radioiodine/contrast material to assess breast neovascularity and provides both anatomical and local changes in the breast. Some trials have now looked into the role of CEM as a tool for screening high-risk younger patients. This has been hailed as an alternative to breast MRI and is cheaper but has a higher radiation dosage .The role of ultrasound elastography has now been extended into identifying breast lesions. It uses ultrasound waves to generate tissue displacement or deformation images in response to external mechanical compression or vibration. These images are then analysed to assess the stiffness or elasticity of the breast tissue. Using a shear wave or strain wave pattern, one can differentiate a benign lesion from a malignant one. These techniques complement each other and work on the basic principle that focal breast malignant lesions are stiffer than benign lesions .Guidelines recommending the use of elastography for characterizing breast lesions have been published by the European Federation of Societies for Ultrasound in Medicine (EFSUMB) and the World Federation of Ultrasound in Medicine and Biology (WFUMB) . Both thBreast localization techniques have been known to guide surgical excisions for non-palpable or occult breast lesions using clips and wires. However, more sophisticated localization techniques have been developed, which can be deployed in advance and cause minimal tissue trauma or migration issues.®, Radiofrequency Iodine seed (RFID) localization, Savi Scout®, and Pintuition® to assist in precisely excising tissue of interest in theatre; this coupled with the use of 3D intraoperative X-rays have greatly helped to reduce breast margin re-excision rates and improved cosmetic outcomes, leading to better patient satisfaction and reduced readmission rates. A significant advantage is the ease of inserting these localizers under local anaesthesia in outpatient settings under ultrasound guidance, and this can be kept in the breast for six months, which can help in surgical scheduling and resource optimization [Increasing use of breast conservation surgery, especially after downstaging following neoadjuvant chemotherapy (NACT), has led to an increase in the use of novel techniques such as Magseedmization ,11,12,13The management of breast lesions with uncertain malignant potential, often called B3 lesions, is slightly complex. These lesions include lesions such as flat columnar cell atypia, atypical ductal hyperplasia (ADH), atypical intraductal epithelial proliferation (AIDEP), radial scar, papillary lesions, and lobular carcinoma in situ (LCIS) . These lAccording to DNA gene profiling, breast cancer can be divided into four subtypes: Luminal A, Luminal B, HER2 enriched, and Triple negative. Multiple studies have shown that the subtypes of ER+ tumours, luminal A and B, have two distinct clinical courses. Of all breast cancer subtypes, luminal A tumours have the best prognosis, whereas luminal B, HER2-enriched, and basal subtypes are associated with poorer clinical outcomes . PatientThe human epidermal growth factor receptor 2 (HER2), or HER2/neu and ERBB2, encodes a transmembrane tyrosine kinase receptor that binds to its extracellular signal, initiating a cascade that mediates cell proliferation, differentiation, and survival. Between 12% and 20% of all breast cancers overexpress the HER2 protein . Other tAccording to the Association of Breast Surgeons (ABS) and the British Association of Plastic, Reconstructive and Aesthetic Surgeons guidelines (BAPRAS), oncoplastic breast surgery should be considered in all patients with breast cancer .These discussions include volume replacement techniques, such as level 2 oncoplastic procedures and perforator flaps, and discussing the role of other reconstruction methods, such as implant-based or autologous flaps following mastectomy, as an immediate or delayed procedure .Following the publication of Keogh’s Review of Regulations of Cosmetic Interventions in the United Kingdom, the Breast and Cosmetic Implant Registry (BCIR) was opened in 2016 . The regImplementing the Enhanced Recovery Programme in breast surgery has revolutionized breast surgery and improved its outcomes . Using aThe use of novel oncoplastic techniques to reduce mastectomy rates wherever possible has provided good cosmetic outcomes and has been the mainstay of surgical treatment for breast cancer recently. The use of techniques such as mammoplasties and local advancement perforator flaps such as lateral thoracic artery perforator (LTAP), lateral intercostal arteries (LiCAP), anterior intercostal arteries (AiCAP), thoracodorsal artery perforator (TDAP), and medial intercostal arteries (MiCAP) have all now become new standards in oncoplastic breast surgery, providing patients with more options for treatment and enabling breast conservation ,32.® and Verita, can provide immediate reconstruction with an implant. This has enabled surgeons to perform the procedure in the prepectoral space without disrupting the pectoralis muscle, which causes less post-operative pain and shoulder problems and provides good cosmetic outcomes. However, with the increased use of implants and adoption of newer cohesive breast implants, the risks of breast implant illness and low-grade anaplastic lymphoma (BIA-ALCL), with an estimated risk of 1:25,000 to 30,000 have now been added to the list of complications with the use of breast implants. These patients are informed about this while deciding on surgery [In patients where breast conservation is not an option, mastectomy is indicated, such as high-risk/BRCA genetic patients, and the choice of breast reconstruction is offered where feasible. Newer techniques, such as Prepectoral breast reconstruction using Acellular Dermal Matrix ,34, such surgery ,36.With the increased use of breast reconstruction, modifications in the management of implant handling during surgery to reduce surgical site infection complications have also become paramount. Using theatre checklists for implants has become a standard of care in hospitals carrying out implant-based reconstruction .During the COVID-19 pandemic, the management of patients with breast cancer who needed radiotherapy changed significantly. Patients who needed radiotherapy were offered a Fast Forward regime. It was established that treating patients with a 26 Gy in five fractions regimen over one week was non-inferior to the standard of 40 Gy in 15 fractions over three weeks for local tumour control. It was safe in terms of normal tissue effects for up to 5 years for patients prescribed local adjuvant radiotherapy after primary surgery for early-stage breast cancer .There is emerging evidence that exercise therapy following breast cancer treatment significantly reduces the chances of recurrence and mortality among breast cancer patients and improves psychological well-being .A meta-analysis of six studies looking at 12,108 breast cancer patients concluded that post-diagnosis physical activity reduced breast cancer deaths by nearly 34% and all-cause mortality by 41% . HoweverA combination of immunotherapy and chemotherapy has recently emerged as a novel treatment option, with encouraging results observed with the combination of immune checkpoint blockade with diverse biological agents, including anti-HER2 agents, cyclin-dependent kinase (CDK) 4/6 inhibitors, and PARP inhibitors. Currently, three selective CDK4/6 inhibitors  have been approved by both the Food and Drug Administration (FDA) and the European Medicines Agency (EMA) for treating HR-positive, HER2-negative metastatic breast cancer . TreatmeThe evolution of AI in breast surgery had progressed from the early stages, when researchers and clinicians started exploring the potential of AI in analysing medical images, such as mammograms, to aid in breast cancer diagnosis and screening. Basic machine learning algorithms were used for image analysis and feature extraction, but the applications were limited. As technology advanced, more sophisticated deep learning algorithms, such as convolutional neural networks (CNNs), showed promise in detecting breast lesions and classifying breast cancer subtypes more accurately. These advances led to more applications of AI in breast surgery, including image-guided surgery, surgical planning, and prediction of treatment outcomes. Currently, numerous studies and clinical trials are exploring the use of AI in various aspects of breast surgery, including image analysis, surgical planning, decision support, and prediction of patient outcomes. AI-powered tools, such as computer-aided detection (CAD) systems, are increasingly used in clinical practice to assist radiologists in breast cancer screening and diagnosis. Virtual reality (VR) and augmented reality (AR) technologies are also being utilized to enhance surgical visualization and training ,4.The ongoing research and development pave the way for the availability in the near future of automated histopathology analysis, prediction of treatment response, and patient-specific treatment planning and optimized surgical approaches.To resurface from the suspension of breast cancer screening during the COVID-19 infection, various centres across the United Kingdom have opted to participate in or conduct clinical trials into using AI as a screening aid for detecting breast cancer. One such centre in England, Leeds, recently announced the use of AI software as a LIBRA study . The aimReliance on AI will likely proceed through stages. It will involve careful attention to mitigate its limitations and challenges around patient privacy, accountability, bias, informed consent, and economic and social justice .Similarly, in digital pathology, transforming histopathology slides into digital images using whole-slide scanners and subsequent analysis of these digitized images by using AI appears promising. It can help speed up the diagnostic process with a limited workforce .The role of the vaccine in breast cancer appears promising and is based on the hypothesis of immunological response via either T or B cells to destroy cancer cells. There have been several trials, but none have been promising until recently. Various types of vaccines are under trial, including peptide-based, DNA-based, tumour cell antigen-based, and carbohydrate-based vaccines . ClinicaVarious alternative treatment methods have been investigated recently, such as microwave ablation of breast lesions , cryotheA useful clinical marker for breast cancer subtype classification, prognosis, and prediction of therapeutic response. This proliferation marker has been adopted in various centres to aid breast cancer treatment. Ki67 is a valuable tool in assessing the risk of recurrence for ER-positive human epidermal growth factor receptor 2 (HER2)-negative breast cancers, where it may be considered a surrogate of molecular assays for distinguishing luminal A from luminal B breast cancer subtypes. Two important clinical trials of NETs, the Immediate Preoperative Anastrozole, Tamoxifen, or Combined with Tamoxifen (IMPACT) trial and P024, established Ki67 as the evaluation index of NETs. IMPACT compared the efficacy of NET with anastrozole, tamoxifen, and a combination of anastrozole and tamoxifen in postmenopausal women with ER-positive invasive primary breast cancer. P024 compared letrozole with tamoxifen in NET ,51,52.The use of complex algorithms and diagnostic modalities to assess predictive and prognostic biomarkers is central to quality oncology care. Advances in sample preparation, microscopy, and image analysis have enhanced immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) methods used in HER2 borderline results. Bright-field ISH methods that can be performed using standard light microscopy, such as chromogenic ISH (CISH), silver ISH (SISH), and dual ISH (a combination of CISH and SISH techniques), have also been developed to overcome the limitations of dark-field fluorescence microscopy and the lack of morphologic details associated with FISH.Additionally, digital microscopy and image analysis technologies are becoming increasingly important tools in pathology. For example, computerized image analysis has been shown to help improve the accuracy and reliability of IHC computerized image analysis. Imaging platforms such as the Aperio ScanScope  and the Definiens  Tissue Studio allow the scanning of whole glass slides to create a digital image that can be analysed using specific algorithms within a short period and can detect regions of interest and distinguishes cells and subcellular attributes within the target regions .Various gene assay techniques have now been used to manage breast cancer care: one such test is the Oncotype Dx  test which uses a 21 gene assay score using a quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay that measures the expression of 21 genes . The results tell us how likely a cancer is to grow and respond to treatment. Therefore, the Oncotype DX Breast Recurrence Score Test is both a prognostic test since it provides more information about how likely (or unlikely) the breast cancer is to come back and a predictive test since it predicts the likelihood of benefit from chemotherapy or radiation therapy treatment . The rolRadiotherapy is often used to treat breast cancer. Still, it has its limitations, such as cardiopulmonary side effects and difficult-to-treat hard-to-reach areas such as internal mammary lymph nodes, which lie close to the heart and lungs . With imWhile there have been significant advancements in breast cancer care, several challenges have arisen along with these innovations. Some of the difficulties associated with these advancements include ,3,4:Cost and Accessibility: Many innovative technologies and treatments in breast cancer care can be expensive, making them less accessible for some patients due to financial constraints or lack of insurance coverage. This can create disparities in access to care, with some patients unable to benefit from the latest advancements due to financial limitations.Ethical and Social Implications: As personalized medicine and genomic testing become more prevalent in breast cancer care, ethical and social implications can arise, such as concerns about genetic privacy, discrimination based on genetic information, and potential psychological impacts of knowing one’s genetic predisposition to breast cancer. These challenges require careful consideration of ethical and social implications to protect patient’s rights and well-being.Integration and Implementation: Integrating and implementing new technologies and treatments into routine clinical practice can be complex and challenging. Healthcare providers may have a learning curve to become proficient in using new technologies and logistical challenges regarding equipment, training, and infrastructure needed for widespread adoption.Evidence-based Decision Making: While advancements in breast cancer care have brought about new treatment options, there may be limited evidence regarding their long-term safety and efficacy. This can make it challenging for healthcare providers and patients to make informed decisions about the best treatment approach, particularly in rapidly evolving fields such as immunotherapy and targeted therapies.Patient Education and Informed Consent: Patient education and informed consent become crucial with the increasing complexity of breast cancer care. Ensuring that patients are adequately informed and able to make informed decisions can be challenging in the context of rapidly evolving advancements. Patients must understand different treatment options’ risks, benefits, limitations, and potential implications of genetic testing and personalized medicine.Health Disparities: Despite the advancements in breast cancer care, there may be disparities in access to these innovations, particularly among underserved populations and minority communities. Addressing health disparities and ensuring equitable access to advancements in breast cancer care remains a challenge that requires attention and action.We believe that through our paper, we can highlight the role of various innovations aimed at improving the ease and precision of detection, personalizing treatment, predicting response to breast care treatment, and improving overall survival. Novel advancements, however, come with newer and unique limitations and challenges. Continued research, advocacy, and bold efforts are essential to address these challenges to make these innovations accessible to all patients in an ethical, equitable, and safe environment.


This bending increases the number and types of dynamic assemblies that can be generated, as well as the spectrum of their functions. Intriguingly, in some cases, actin bending creates directionality within a cell, resulting in a chiral cell shape. This actin-dependent cell chirality is highly conserved in vertebrates and invertebrates and is essential for cell migration and breaking L-R symmetry of tissues/organs. Here, we review how different types of actin binding protein can bend actin filaments, induce curved filament geometries, and how they impact on cellular functions.In many cellular contexts, intracellular actomyosin networks must generate directional forces to carry out cellular tasks such as migration and endocytosis, which play important roles during normal developmental processes. A number of different actin binding proteins have been identified that form linear or branched actin, and that regulate these filaments through activities such as bundling, crosslinking, and depolymerization to create a wide variety of functional actin assemblies. The helical nature of actin filaments allows them to better accommodate tensile stresses by untwisting, as well as to bend to great curvatures without breaking. Interestingly, this latter property, the bending of actin filaments, is emerging as an exciting new feature for determining dynamic actin configurations and functions. Indeed, recent studies using  Xenopus convergent extension, Zebrafish epiboly, C. elegans ventral enclosure, Drosophila germband extension/retraction and dorsal closure, and mouse palate formation15. The generation of such intracellular forces depends heavily on the dynamic organization and regulation of actomyosin networks and other cytoskeletal proteins. Actin and myosin form a number of different functional assemblies within cells, including supracellular actomyosin cables, cortical-junctional actomyosin, apical-medial actomyosin, and basal actomyosin networks21. Actin filaments and myosin form distinct dynamic structural geometries within these different assemblies based of their spatial organization and connectivity, which leads to different proficiencies in force generation21.Directional forces within cells are necessary for a wide spectrum of developmental events including cytokinesis, left/right (L-R) asymmetry, de novo linear actin nucleation factors such as formins and Spire25  depending on the physical conformation of the actin filament42. Intriguingly, a recent study showed that the nucleotide state of actin filaments can have a major impact on the rigidity of the filament and influence its twisting and bending42. The specific nucleotide state in the presence of bending forces can additionally influence the spatial rearrangements of actin molecules and change the helical twist of actin filaments as predicted by computer simulations43. Therefore, there exists a complex feedback loop between the physical stresses applied to an actin filament, its conformation, and the combination of ABPs bound. This complex system of forces and ABPs can dictate the conformation of a given actin filament and/or the organization/geometry of actin filament assemblies to properly sense and react to cellular and environmental events.Actin filaments themselves are also inherently helical and this helicity has been demonstrated to provide the filament with its mechanical strength and play an important role in facilitating cell chirality and L-R asymmetric morphogenesis34 . InteresIn addition to their roles in assembling linear and branched actin filament assemblies, a number of ABPs have recently been shown to alter the properties of actin filaments, triggering them into curved geometries. Here, we review the ability of different ABPs—alone or in conjunction with other ABPs—to alter the conformation of actin filaments and/or the geometry of actin filament assemblies, and their effect on cellular and developmental events.46. IQGAPs possess recognizable protein motifs, including calponin homology domains (CH), coiled-coil domains (CC), and GTPase R domains (GRD), that contribute to their modulation of actin assemblies45. The CC domain has been demonstrated to bind to Ezrin, a cytoskeleton-plasma membrane anchor, and the GRD domain transiently protects the Cdc42 small GTPase in its activated state by reducing GTP hydrolysis. These two domains allow the IQGAP family to localize to the plasma membrane and sustain the interaction between the WASP branched actin nucleator and the Arp2/3 complex through its CH domain, thereby promoting branched actin nucleation47. In addition to binding WASP, the CH domain has been shown to bind actin filaments with high affinity and, strikingly, to bend actin filaments to a high curvature48. In a recent study, Palani and colleagues creatively used the CH domain of Rng2 (fission yeast IQGAP ortholog) to show that it was capable of bending actin filaments to form ring structures48  and other actin binding proteins  to show that the conformation of the actin filament can alter how well the reporter bound to actin filaments. As actin filaments are inherently helical, some bound proteins can change the pitch of actin and induce twisting or bending forces to give rise to high-order actin assemblies such as a ring. The bending or twisting of actin filaments can sterically influence the localization of other ABPs or result in preferential binding of specific ABPs 38. Thus, there is a striking reciprocity in which the binding of ABPs alters the conformation of actin filaments, while the conformation of actin filaments affects which ABPs can bind.The IQGAP (IQ motif-containing GTPase-activating proteins) protein family are scaffolding proteins with multifarious interactions that allow them to facilitate actin dynamics at the cell membrane. They have been implicated in dynamic actin processes such as cell migrationctures48 . In a se53. Notably, Cofilin has been shown to sever bent actin filament regions more efficiently54. When Cofilin binds to actin filaments, the helical half-pitch of the decorated actin filaments (referred to as “cofilactin” filaments) changes: 37 nm for bare actin filaments and ~27 nm for cofilactin filaments57. The shorter helical pitch and the transition from bare to cofilactin filaments led to accelerated actin severing. However, saturated cofilactin filaments are no longer severed cofilin-dependently and are more flexible58. While examining actin bundles within filopodia at neuronal growth cones using cryo-electron tomography, Hylton and colleagues recently showed that cofilactin filaments associate with curvature of the actin bundles, thereby regulating filopodia structure and dynamics59. Interestingly, this cofilactin filament conformational change results in an exclusion of the Fascin actin-crosslinking protein from cofilactin bundles, leading to less organized actin filament bundling, the bending of filopodial protrusions, and a disruption of filopodial structure and functions , Anillin may do more than simply bundling actin: Anillin may act to generate and facilitate organizational changes in Septins and actin to modulate the tunability and optimization achieved by actin assemblies.Anillin is a crosslinker of both Septins and actin that contains a pleckstrin homology domain allowing it to anchor to cell membranest manner 73. Simil83. Actin crosslinkers can dynamically regulate the stiffness of actin filament bundles and the stability of actin networks through bundling or bridging actin filaments in many cellular processes, including filopodia formation and focal adhesion assembly86. While both α-Actinin and Fascin can bundle individual actin filaments to form tightly packed parallel filaments, the structures of these resulting actin filament bundles are different. A combination of TEM and in vitro actin bundling assays showed that actin filaments bundled by α-actinin have ~35 nm space between each filament and form square lattice structures, whereas actin filaments bundled by Fascin have ~8 nm space and form a hexagonal lattice structure83  asymmetric morphogenesisd/buckle .105. Their conserved N-terminal motor domains bind to actin and their class-specific C-terminal domains regulate the selection of and binding to cargos or other cell components105. Unconventional myosins have been studied with a focus on their function as cargo transporters, molecular anchors (to the actin cytoskeleton or plasma membrane), tension sensors, actin crosslinkers, signal transducers, and mediators of membrane-cytoskeleton interactions needed for a wide range of cellular processes, including endo-/exo-cytosis, organelle and membrane trafficking, cell motility, and L-R asymmetry106. Type I unconventional myosins are tethered to the membrane and regulate cellular processes such as membrane tension and intracellular trafficking in cells107. While examining the role of type I unconventional myosins in the generation of L-R asymmetry in different Drosophila tissues, Lebreton and colleagues eloquently showed that two of these, Myo1D and Myo1C, generate de novo directional twisting of cells and organs in opposite directions to establish chirality110. Intriguingly, the motor domain of Myo1D, but not that of Myo1C, produced counterclockwise circular motility of actin filaments in vitro. In contrast to Drosophila Myo1C, mammalian Myo1C does produce a counterclockwise circular motility of actin filaments in vitro , provides regions for more efficient severing54, and can be used to generate force by unbending. In addition, a recent study suggests that nucleotide-dependent modifications of the actin filament conformational landscape influences the binding specificity of ABPs42. Intriguingly, α-Actinin, IQGAP, and unconventional myosins also induce directional actin filament movements such as counterclockwise swirling. This actin-dependent cell chirality is observed in a wide range of animals from Drosophila to mammals121. While much of the evidence for actin bending and its biological consequences to date comes from studies using in vitro systems, recent studies using cells and model organisms revealed that cell chirality has important roles in processes such as cell migration and L-R asymmetric organ development123. We still have much to learn about how altering the conformation of actin filaments by bending them into curved geometries affects the functions of actin assemblies, results in chirality, and the means by which it is used to achieve precise developmental outcomes. In addition, it remains unclear how ABP binding induces actin filament bending and directional movement of actin filaments. The combination of cryo-EM, super-resolution microscopy, cutting-edge live-imaging techniques, single-cell RNAseq, and computer simulations can aid in further understanding the mechanistic insights and revealing the mysteries still surrounding the role of actin bending.Actin networks must dynamically change their configurations to generate the forces needed within cells for many of their essential biological processes