Allogeneic Tissue Research Articles

Locally controlled release of immunosuppressive promotes survival of transplanted adult spinal cord tissue.

Transplantation of adult spinal cord tissue (aSCT) is a promising treatment for spinal cord injury (SCI) basing on various types of neural cells and matrix components inside aSCT. However, long-term systemic administration of immunosuppressors (e.g. tacrolimus, TAC) is required for the survival of allogeneic tissue, which often associated with severe side effects such as infection, liver damageand renal failure. In this study, a triglycerol monostearate (TGM)-based TAC delivery system (e.g. TAC@TGM) with high drug loading concentration was developed, which possessed injectable properties as well as sustainable and immune-responsive drug release behaviors. In complete transected SCI model, locally injected TAC@TGM could reduce the infiltration of inflammation cells, enhance the survival of transplanted aSCT (e.g. Tuj-1+ and NF+ neurons) and promote the recovery of locomotor function. Moreover, controlled release of TAC by TAC@TGM attenuated side effects of TAC on liver and kidneys compared with traditional systemic administration. More importantly, the developed TAC@TGM system provided a facile single dose of long-term immunosuppressive effect not just for aSCT transplantation, but also for other tissue/organ and cell transplantations.

Polymeric Materials for Bone and Cartilage Repair Using IBM SPSS Statistics Method

Polymeric materials for bone and cartilage repair Introduction: Polymers are called macromolecules made up of very large molecules natural or artificial one of the classes of objects, the simple chemicals called monomers are multiples of units. Polymers many substances in living things make, for example, proteins, cellulose, and nucleic acids. Natural and synthetic polymeric materials in bone tissue engineering are widely used because they are extracellular similarity to matrices and have considerable biocompatibility and biodegradability. Research significance: One of the most harmful and expensive issues in human health is dysfunction, injuries, or other types of damage to human tissue and organs. Different surgical procedures have been developed to solve these concerns, including total artificial substitutes, combined artificial kidneys, etc. Inert processed tissues heart valves, autogenic or allogeneic tissues, and total artificial kidney and prosthetic kidneys (transplantation). Organ transplantation will face significant challenges due to the scarcity of organ donors, the lifetime requirement for immunity, and its devastating consequences. Also, transplanted tissue does not provide all the functionality own tissue and can covers the donor site problem. Concerning the non-biological, there could be material issues. Like, disease, and shortage limited durability and biocompatibility. These factors have made stem cells, tissue engineering (te), and organogenesis promising and significant industries. Research doesn't offer anything. Just organs and tissues transplantation, however, can provide fresh viewpoints medications for illnesses Methodology: SPSS statistics is a data management, advanced analytics, multivariate analytics, business intelligence, and criminal investigation developed by IBM for a statistical software package. A long time, spa inc. Was created by, IBM purchased it in 2009. The brand name for the most recent versions is IBM SPSS statistics. Evaluation parameters: Natural Polymer, Proteins Collagen, Silk fibroin, Polysaccharides Chitosan, Alginates, Bacterial cellulose, Synthetic Polymers. Results: the Cronbach's Alpha Reliability result. The overall Cronbach's Alpha value for the model is .590 which indicates 59% reliability. From the literature review, the above 61% Cronbach's Alpha value model can be considered for analysis. Conclusion: Cronbach's alpha reliability results. The overall Cronbach's alpha value for the model was .590, indicating a reliability of 59%. From the literature review, the above Cronbach's alpha value of 61% can be considered to analyze the model.

Developing autologous cell therapy for macular degeneration: From bench‐to‐bedside

AbstractInduced pluripotent stem cells (iPSCs) can provide autologous and allogeneic replacement tissues, potentially for all degenerative diseases. Autologous tissues have the advantage of not requiring immune‐suppressive drugs that have deleterious side‐effects. The safety and feasibility of autologous iPSC‐based therapies has not been established. Here, we developed an autologous iPSC‐based therapy for age‐related macular degeneration (AMD), a blinding eye disease that affects over 30 million people world‐wide. AMD is caused by the progressive degeneration of retinal pigment epithelium (RPE), a monolayer tissue that maintains photoreceptor function and survival. We tissue engineered a clinical‐grade iPSC‐RPE‐patch on a biodegradable scaffold using autologous cells from AMD patients. Preclinical investigational new drug (IND)‐enabling studies performed on iRPE‐patch derived from multiple AMD patients demonstrated reproducible manufacturing, validating our manufacturing process – a key requirement for an autologous phase I trial. Functional validation of clinical‐grade iRPE‐patches allowed determination of tissue barrier resistance, purity of RPE cells, and RPE cell shape metrics as key critical quality attributes now used as clinical product release criteria. Preclinical animal studies performed in immune‐compromised rats confirmed safety of the auto‐iRPE‐patch and efficacy studies performed in a porcine laser‐induced RPE injury model that mimics AMD‐like eye conditions demonstrated integration and functionality of RPE patches. A phase I/IIa IND‐application for an auto‐iRPE‐patch to treat AMD was recently cleared by the FDA. This Phase I/IIa clinical trial will test safety, feasibility, and integration of an auto‐iRPE‐patch in twelve advanced AMD patients. The trial is currently being run as a single site study at the NIH and targets patient enrollment in two cohorts – with visual acuity between 20/100 to 20/500 and between 20/80 to 20/500.

Physicochemical evaluation of the acellular tracheal graft for tissue remodeling

In a biological system, the airway system upper respiratory tract, or trachea connects the nostrils and lungs, to help in breathing process and trapping dust and pathogen particles. Clinically, to overcome the tracheal defects and dsyfunctioning different type of artificial prosthetics, stents, and grafts are used to reconstruct tracheal tissue at the injured tissue. Decellularized tissue fabricated from xenogeneic and allogeneic tissue are Extracellular matrix (ECM) protein rich, biocompatible, 3D biomimetic grafts having potential mechanical stability for better tissue regeneration. Acellular tracheal graft fabricated from the cadaver goat-tracheal tissue have 3D interconnected porous structure with good mechanical stability upto 18 MPa measured by UTM, stability and control degradability rate. Biochemical analysis by the FTIR spectra analysis and assays confirmed the presence of stable collagenous protein triple helix structure with retained majority of the ECM (extra cellular matrix) molecules (collagen, proteoglycans, elastin and GAGs) without any significant loss, bio-degradability, and physiochemical stability.

Multifunctional hydrogel modulates the immune microenvironment to improve allogeneic spinal cord tissue survival for complete spinal cord injury repair

Transplantation of allogeneic adult spinal cord tissues (aSCTs) to replace the injured spinal cord, serves as a promising strategy in complete spinal cord injury (SCI) repair. However, in addition to allograft immune rejection, damage-associated molecular pattern (DAMP)-mediated inflammatory microenvironments greatly impair the survival and function of transplants. In this study, we aimed to regulate the immune microenvironment after aSCT implantation by developing a functional hybrid gelatin and hyaluronic acid hydrogel (F-G/H) modified with cationic polymers and anti-inflammatory cytokines that can gelatinize at both ends of the aSCT to glue the grafts for perfect matching at defects. The F-G/H hydrogel exhibited the capacities of DAMP scavenging, sustainably released anti-inflammatory cytokines, and reduced lymphocyte accumulation, thereby modulating the immune response and enhancing the survival and function of aSCTs. When the hydrogel was used in combination with a systemic immunosuppressive drug treatment, the locomotor functions of SCI rats were significantly improved after aSCTs and F-G/H transplantation. This biomaterial-based immunomodulatory strategy may provide the potential for spinal cord graft replacement for treating SCI. Statement of significanceIn this study, we aimed to regulate the immune microenvironment by developing a functional hybrid gelatin and hyaluronic acid hydrogel (F-G/H) modified with cationic polymers and anti-inflammatory cytokines that can gelatinize at both ends of the aSCT to glue the grafts for perfect matching at defects. We found that with the treatment of F-G/H hydrogel, the aSCT survival and function was significantly improved, as a result of reducing recruitment and activation of immune cells through TLR- and ST-2- related signaling. With the combination of immunosuppressive drug treatment, the locomotor functions of SCI rats were significantly improved after aSCTs and F-G/H transplantation. Findings from this work suggest the potential application of the F-G/H as a biomaterial-based immunoregulatory strategy for improving the therapeutic efficiency of the transplanted spinal cord graft for spinal cord injury repair.

A Survey of 1000 Respondents on the Polish Population's Knowledge and Attitudes about Tissue/Organ Donation and Transplantation in Times of Allogeneic Tissue Shortage.

Tissue-engineered human allogeneic skin grafts retrieved from a deceased donor play an important role in the therapy of extensive and deeply burned patients. However, there is a vital deficit of allogeneic skin donors, and the reserves of human allogeneic skin grafts are not sufficient. The goal of this work was to analyze the level of knowledge and attitudes of Polish society in the field of transplantation, with particular emphasis on allogeneic skin transplantation. The study used a self-made questionnaire comprised of 23 questions. 1000 respondents took part in this research. The respondents were a diverse group in terms of age, sex, education, and place of residence. The obtained results show a general positive attitude of the respondents towards the idea of transplantology. However, people with lower education presented a more negative attitude towards the donation of tissues and organs. Additionally younger people were not able to clearly declare readiness for organ procurement. What is more data analysis revealed certain gaps in more detailed knowledge and surprising attitudes. In that respect, the lack of awareness about the criteria for determining brain death could be mentioned. There was also a lack of acceptance for skin procurement in specific population groups. It can therefore be concluded that a key role in the success of the idea of transplantation in Poland is the broad and systematic education of the society.

Allogeneic osteoplastic materials for reconstructive surgery of combat injuries

The number of operations with the use of osteoplastic materials is increasing annually, which is explained by the increasing number of high-tech operations, reconstructive interventions for bone pathology, oncoorthopedics, bone deformities, as well as tissue defects resulting from combat trauma. Bone as an object of transplantation and is preferable because the use of bone tissue and materials made on its basis creates the necessary conditions for biological restoration of bone as an organ.
 The modern trend is the development of regenerative medicine technologies and the development of modified materials with improved and predetermined properties osteoplastic materials ceased to be static structures and their inertness gave way to functional activity.
 The article describes the basic advantages of bone tissue and allogenic osteoplastic materials for their prospective use in reconstructive and reconstructive surgery of the musculoskeletal system. Data on the experience in the use of donor bone tissues for homoplasy at the clinic of military traumatology and orthopedics of the Kirov Military Medical Academy are presented. The advantages of allogeneic decellularized tissues for use in clinical and scientific practice are considered. The importance of preserving the architecture of native tissue for successful bone transplantation is shown. Decellularization is a method that allows the removal of immunogenic agents from tissues and organs, including cells and residual DNA, preserving the natural composition and architecture of the extracellular matrix for the most effective use of allogeneic bone tissue. The use of decellularized allogenic bone tissue purified to a mineral-collagen matrix or demineralized is the most practical option among other osteoplastic materials when large bone fragments need to be replaced.

Результаты хирургического лечения птеригиума с применением двухкомпонентного аутофибринового клея

The only effective way to treat pterygium is to perform surgical intervention in various ways (without replacing the defect of the conjunctiva and replacing the defect with allogeneic or autologous tissues). One of the most effective methods is considered to be the removal of the pterygium with defect plasty with an amniotic membrane. The issue of choosing the fixation of the amniotic membrane is topical. The use of a suture fixation method is considered the gold standard, but this method increases the duration of the operation, and is also accompanied by discomfort for the patient in the postoperative period. The use of an adhesive fixation method using autologous fibrin adhesives is considered promising. Purpose of the study: to conduct a comparative analysis of the effectiveness of surgical treatment of pterygium using a two-component autofibrin glue and sutures for fixing the amniotic membrane. Material and methods. The study included 15 patients. All patients were divided into 2 groups depending on the method of fixing the amniotic membrane: the 1st group was the main group, where the amniotic membrane was fixed with a two-component autofibrin glue, the 2nd group was the control group, where the amniotic membrane was fixed with sutures. All patients underwent biomicroscopy and optical coherence tomography of the anterior segment to determine the stage of pterygium. Before surgical treatment, tear production was assessed using the Schirmer test. The study groups compared the time spent on the operation. After the surgical intervention, the control periods were followed by photo registration, optical coherence tomography of the anterior segment, as well as an assessment of the presence of the following "symptoms" in the patient: pain, sensation of a foreign body, itching, lacrimation, assessed in points on a visual analogue scale. After 6 months, the presence of pterygium recurrence in the study groups was assessed. Statistical analysis was carried out using the StatTech v. 2.8.8. Results. As a result of evaluating the indicator "Duration of surgery" depending on the study group, we found statistically significant differences (p = 0.032). When assessing the "Pain" indicator, statistically significant differences were detected on the 1st (p = 0.007), 3rd (p = 0.03) and 7th days (p = 0.047) after the operation. When assessing the indicators of the presence of "Foreign body sensation" and "Itching", statistically significant differences between the groups were detected only on the 1st day after the operation (p = 0.04 and p = 0.017, respectively), in the remaining control periods, differences between the groups were revealed did not have. When analyzing the indicator of "Lacrimation", no statistically significant differences between the studied groups were revealed for all control periods of the study. No recurrence of pterygium was recorded in any of the cases 6 months after the surgical interventions. Conclusion. Fixation of the amniotic membrane with the help of the developed two-component autofibrin glue during the surgical treatment of pterygium provides not only a reduction in the operation time, but also a more comfortable postoperative period. Keywords: pterygium, autofibrin glue, surgical treatment, amniotic membrane.

Optimal therapeutic conditions for the neural stem cell-based management of ischemic stroke: a systematic review and network meta-analysis based on animal studies

BackgroundIn order to promote the clinical translation of preclinical findings, it is imperative to identify the most optimal therapeutic conditions and adopt them for further animal and human studies. This study aimed to fullyexplore the optimal conditions for neural stem cell (NSC)-based ischemic stroke treatment based on animal studies.MethodsThe PubMed, Ovid-Embase, and Web of Science databases were searched in December 2021. The screening of search results, extraction of relevant data, and evaluation of study quality were performed independently by two reviewers.ResultsIn total, 52 studies were included for data analysis. Traditional meta-analysis showed that NSCs significantly reduced the modified neurological severity score (mNSS) and volume of cerebral infarct in animal models of ischemic stroke. Network meta-analysis showed that allogeneic embryonic tissue was the best source of NSCs. Further, intracerebral transplantation was the most optimal route of NSC transplantation, and the acute phase was the most suitable stage for intervention. The optimal number of NSCs for transplantation was 1–5×105 in mouse models and 1×106 or 1.8×106 in rat models.ConclusionsWe systematically explored the therapeutic strategy of NSCs in ischemic stroke, but additional research is required to develop optimal therapeutic strategies based on NSCs. Moreover, it is necessary to further improve and standardize the design, implementation, measuring standards, and reporting of animal-based studies to promote the development of better animal experiments and clinical research.

Tissue Engineering Neovagina for Vaginoplasty in Mayer-Rokitansky-Küster-Hauser Syndrome and Gender Dysphoria Patients: A Systematic Review.

Background: Vaginoplasty is a surgical solution to multiple disorders, including Mayer-Rokitansky-Küster-Hauser syndrome and male-to-female gender dysphoria. Using nonvaginal tissues for these reconstructions is associated with many complications, and autologous vaginal tissue may not be sufficient. The potential of tissue engineering for vaginoplasty was studied through a systematic bibliography search. Cell types, biomaterials, and signaling factors were analyzed by investigating advantages, disadvantages, complications, and research quantity. Search Methods: A systematic search was performed in Medline, EMBASE, Web of Science, and Scopus until March 8, 2022. Term combinations for tissue engineering, guided tissue regeneration, regenerative medicine, and tissue scaffold were applied, together with vaginoplasty and neovagina. The snowball method was performed on references and a Google Scholar search on the first 200 hits. Original research articles on human and/or animal subjects that met the inclusion (reconstruction of vaginal tissue and tissue engineering method) and no exclusion criteria (not available as full text; written in foreign language; nonoriginal study article; genital surgery other than neovaginal reconstruction; and vaginal reconstruction with autologous or allogenic tissue without tissue engineering or scaffold) were assessed. The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist, the Newcastle-Ottawa Scale, and the Gold Standard Publication Checklist were used to evaluate article quality and bias. Outcomes: A total of 31 out of 1569 articles were included. Data extraction was based on cell origin and type, biomaterial nature and composition, host species, number of hosts and controls, neovaginal size, replacement fraction, and signaling factors. An overview of used tissue engineering methods for neovaginal formation was created, showing high variance of cell types, biomaterials, and signaling factors and the same topics were rarely covered multiple times. Autologous vaginal cells and extracellular matrix-based biomaterials showed preferential properties, and stem cells carry potential. However, quality confirmation of orthotopic cell-seeded acellular vaginal matrix by clinical trials is needed as well as exploration of signaling factors for vaginoplasty. Impact statement General article quality was weak to sufficient due to unreported cofounders and incomplete animal study descriptions. Article quality and heterogenicity made identification of optimal cell types, biomaterials, or signaling factors unreliable. However, trends showed that autologous cells prevent complications and compatibility issues such as healthy cell destruction, whereas stem cells prevent cross talk (interference of signaling pathways by signals from other cell types) and rejection (but need confirmation testing beyond animal trials). Natural (orthotopic) extracellular matrix biomaterials have great preferential properties that encourage future research, and signaling factors for vascularization are important for tissue engineering of full-sized neovagina.

Regenerative Medicine to Improve Outcomes of Ventral Hernia Reconstruction (REPAIR Study) Phase 1: Find the Best Decellularization Protocol for the Human Dermis.

Background: Tissue regeneration is a complex process that allows wounds to heal. Many options are currently available to help human skin repair and to reduce the recurrence of hernias. The aim of this study is to analyze the best decellularization protocol for allogenic human dermal tissues. Methods: Dermal flaps from donors were used and compared with a control group. Each flap was subjected to seven different decellularization protocols and washed with a sequence of five solutions. The samples were then subjected to four control tests (such as Nile Red), and long-term contacts were analyzed to assess whether the decellularized dermis samples could support the growth of human fibroblasts. Results: All the samples had an average residual viability of 60%. Except for one sample, the decellularization treatments were able to reduce cell viability significantly. The Nile Red test showed a significant reduction in phospholipid content (mean 90%, p-value < 0.05) in all treatments. The cell growth increased in a linear manner. As described in the literature, sodium-dodecyl-sulfate (SDS) caused an interference between the test and the detergent. Conclusions: This paper shows the first step to finding the best decellularization protocol for allografting human dermal tissues. Further biocompatibility tests and DNA quantification are necessary.

Problems of creating bioartificial organs and the competition of medical technologies

A crucial direction in the development of biotechnology is production of human bioartificial organs and of related products for tissue and organ engineering – constituent components for biofabrications of organs and tissues: human biopolymers of extracellular matrix (ECM), cytokines, biomass of autologous and allogeneic human cells, organoids and tissue spheroids, as well as special equipment – tissue bioreactors, bioprinters, etc. The biotechnology development level today allows creating live ‗spare parts‘ for humans, and there is the prospect of competition with manufacturers of prostheses and implants. The present review considers the unsolved problems of creating bioartificial organs from the perspective of the classical triad of tissue engineering technology – cells, scaffold, and bioreactor. The author focuses on clinical examples and discusses possible strategies and new approaches for growing bioartificial organs inside in vivo bioreactors (hereinafter –regeneratrons) integrated with a living organism. Regeneratrons are the latest generation of tissue bioreactorsrarely used in clinical practice yet with a potential of increasing the efficiency of treatment.

A minimally manipulated preservation and virus inactivation method for amnion/chorion.

Allogeneic amnion tissues have been widely used in tissue repair and regeneration, especially a remarkable trend of clinical uses in chronic wound repair. The virus inactivation procedures are necessary and required to be verified for the clinical use and approval of biological products. Cobalt-60 (Co-60) or electron-beam (e-beam) is the common procedure for virus and bacterial reduction, but the excessive dose of irradiation was reported to be harmful to biological products. Herein, we present a riboflavin (RB)-ultraviolet light (UV) method for virus inactivation of amnion and chorion tissues. We used the standard in vitro limiting dilution assay to test the viral reduction capacity of the RB-UV method on amnion or chorion tissues loaded with four types of model viruses. We found RB-UV was a very effective procedure for inactivating viruses of amnion and chorion tissues, which could be used as a complementary method to Co-60 irradiation. In addition, we also screened the washing solutions and drying methods for the retention of growth factors.

Middle Aged, Everlasting Athletes, and Osteoarthritis: The Present and Future.

One issue was definitely not enough! We are prompted to be active, and we are told that exercise is medicine. Indeed, exercise is great medicine: it keeps at bay metabolic conditions associated with sedentarism, and exerts a favorable effect on our mental health. Some of us are born exercisers, some become so, some drift from being great athletes in youth to a life in an armchair, and some discover exercise later in life. Many of us are competitive, and we often compete against ourselves, never to win. For all of us, however, middle-age looms, and osteoarthritis (OA) may become a major issue: in the West, up to 20% of individuals close to retirement age suffer from it, and their life can be ruined. It is not surprising, therefore, that so much interest is placed on how to prevent, diagnose, manage, “cure” OA: a never-ending battle, with no end in sight. As physicians, we would like to know why some of our patients have developed OA, and others are spared. Why some present little evidence of joint degeneration, and are crippled with pain. Above all, we want to make our patients better. The first part of this series produced a great set of evidence-based systematic reviews of the literature on several epidemiological aspects of OA and middle age sports practice. In a natural evolution, this second issue of Sports Medicine and Arthroscopic Review continues along those lines, and dwells in some aspects of biology and regenerative medicine, examining, for example, how drugs can be delivered in a novel fashion, and whether allogeneic tissues can be used to regenerate our battered articular cartilage. In this field, imagination has to be married with high-level science to progress our clinical knowledge. Eventually, exercise is medicine, and, as with all medications, too much is never too good. The spectre of overdosing must be corrected by our regenerative capabilities. Hopefully, this issue will help pave the way. On the Cover: Role of cytokines in mediating the communication between immune cells. In color, the molecules upregulated in platelet-rich plasma compared with platelet-poor plasma. Sports Med Arthrosc Rev. 2022;30:141–146.

Allogenic Umbilical Cord Tissue for Treatment of Knee Osteoarthritis.

Interest in use of perinatal allogenic tissues including clinical-grade minimally manipulated umbilical cord tissue-derived allograft formulations to treat knee osteoarthritis (OA) patients is increasing. Limited studies have characterized these formulations and evaluated their safety and efficacy in knee OA patients. We developed such formulation and reported the presence of growth factors, cytokines, hyaluronic acid, and exosomes. We reported that its administration is safe, and resulted in 50% pain reduction and improvement in knee injury and osteoarthritis outcome score (over 10%) and 36-item short form survey (25%). Another study reported no adverse events post injection of similar formulation and statistically significant ( P <0.001) improvement in visual analog scale and Western Ontario and McMaster Universities Osteoarthritis Index scores and reduction in medication usage in patients (77.8%). We also summarized the clinical trials registered on ClinicalTrials.gov utilizing umbilical cord tissue for knee OA treatment. In conclusion, available studies are preliminary but pave the way to higher level appropriately powered investigations, and these formulations should be considered as nonoperative alternative to manage knee OA.

Allogeneic testes transplanted into partially castrated adult medaka (Oryziaslatipes) can produce donor-derived offspring by natural mating over a prolonged period

Generally, successful testis transplantation has been considered to require immune suppression in the recipient to avoid rejection of the transplanted tissue. In the present study, we demonstrate in medaka that allogeneic adult testicular tissue will engraft in adult recipients immediately after partial castration without the use of immunosuppressive drugs. The allografted testes are retained in the recipient’s body for at least 3 months and are able to produce viable sperm that yield offspring after natural mating. Some recipients showed a high frequency (over 60%) of offspring derived from spermatozoa produced by the transplanted testicular tissue. Histological analyses showed that allografted testicular tissues included both germ cells and somatic cells that had become established within an immunocompetent recipient testis. The relative simplicity of this testis transplantation approach will benefit investigations of the basic processes of reproductive immunology and will improve the technique of gonadal tissue transplantation.

83. Vitamin D3 Enhances Adipose Stromal Cell Bioenergetics Capacity Andproliferation Under Hypoxic Conditions and Improves Fat Grafting Outcomes

Purpose: Reabsorption of autologous fat grafts is often unpredictable and thus volumetric results are not reliable. We have previously demonstrated the efficacy of vitamin D as a lipoprotectant to enhance adipose tissue survival in vitro, ex vivo, and in xenograft human-to-murine in vivo models. Our preliminary results showed that VD3 primarily affected adipose stromal cells and thus in this study, we evaluated the impact of VD3 on human adipose stromal cell (ASC) proliferation and metabolism. In addition, we tested oral VD3 efficacy in a clinically relevant porcine model of subcutaneous fat grafting. Methods: In vitro, lipoaspirate was either a) enzymatically dissociation to free the stromal vascular fraction (SVF) or maintained in culture as tissue particles for 7-days. For SVF cultures, cells were assessed under normoxic or hypoxic conditions in the presence of VD3 and general metabolic and proliferative indices were measured with Seahorse Bioanalyzer and MTT, respectively. For tissue particles, stromal cells and adipocytes were isolated and viability and size were measured with Calcein/PI cytometry. In vivo, female 3-4-month-old Yorkshire swines were sacrificed to provide fresh adipose which was milled under sterile conditions and injected subcutaneously on the dorsum with an 18-gauge cannula. Recipients were herd matched to allogeneic donors. Each donor was used to provide adipose for a matched pair of recipients. Treatment groups received oral vitamin D post-engraftment and compared to matched-chow controls. Grafts were followed serially with ultrasound. After 8- or 12-weeks post-procedure subjects were sacrificed for terminal adipose weights, volumes, and histology. Results: VD3 improves viability and proliferation of adipose stromal cells (p<0.05, N = 6 unique donors). This effect was most evident under persistent hypoxic stress and remained for up to a week post-harvest (p<0.05). Metabolically, VD3 significantly increased cellular energetic reserve capacity under hypoxia which increases the cellular ATP supply (p = 0.0001 for VD3 concentrations versus control). In vivo, as with prior xenografted murine work we were able to successfully transplant herd-matched allogeneic adipose tissue with maintained survival up to 8-12 weeks post engraftment with oral vitamin D tolerance noted in all cases. Volume retention studies are on-going. Conclusion: We have demonstrated that Vitamin D is a safe and effective option for prevention of fat graft loss in vitro and in mice. Here we perform validation in a clinically translatable porcine model. Further, we have demonstrated hypoxia dependent changes in human adipose stromal metabolism and proliferation with vitamin D supporting a more proliferative and energetic profile in those adipose stem cells necessary for fat graft renewal.

Characterization of Decellularized Implants for Extracellular Matrix Integrity and Immune Response Elicitation.

Biological scaffold is a popular choice for the preparation of tissue-engineered organs and has the potential to address donor shortages in clinics. However, biological scaffolds prepared by physical or chemical agents cause damage to the extracellular matrix (ECM) by potentially inducing immune responses after implantation. The current study explores the fate of the decellularized (DC) scaffolds using a cocktail of chemicals following implantation without using immunosuppressants. Using the syngeneic (Lewis male-Lewis female) and allogeneic (Brown Norway male-Lewis female) models and different tissue routes (subcutaneous vs. omentum) for implantation, we applied in-depth quantitative proteomics, genomics along with histology and quantitative image analysis tools to comprehensively describe and compare the proteins following DC and postimplantation. Our data helped to identify any alteration postdecullarization as well implantation. We could also monitor route-specific modulation of the ECM and regulation of the immune responses (macrophage and T cells) following implantation. The current approach opens up the possibility to monitor the fate of biological scaffolds in terms of the ECM and immune response against the implants. In addition, the identification of different routes helped us to identify differential immune responses against the implants. This study opens up the potential to identify the changes associated with chemical DC both pre- and postimplantation, which could further help to promote research in this direction. Impact Statement The development of a biological scaffold helps in the preparation of a functional organ in the clinics. In the current study, we develop a strategy for chemical decellularization and explored two different routes to understand the differential responses elicited postimplantation. The use of sensitive protein and genomic tools to study the changes creates a favorable environment for similar efforts to develop and characterize biological scaffolds before further trials in the clinics. The current study, which was carried out without any immunosuppressive agents, could help to establish (a) appropriate chemical strategies for preparing biological scaffolds as well as (b) identify putative implantable routes to circumvent any adverse immune reactions, which will ultimately decide the outcome for acceptance or rejection of the scaffold/implant.

Allogenic perinatal tissue for musculoskeletal regenerative medicine applications: a systematic review protocol

BackgroundMusculoskeletal ailments impact the lives of millions of people, and at times necessitate surgery followed by physiotherapy, drug treatments, or immobilization. Regenerative musculoskeletal medicine has undergone enormous progress over the last few decades. Sources of tissues used for regenerative medicine purposes can be grouped into autologous or allogenic. Although autologous sources are promising, there is a wide range of limitations with the treatment, including the lack of randomized controlled studies for orthopaedic conditions, donor site morbidity, and highly variable outcomes for patients. Allogenic sources bypass some of these shortcomings and are a promising source for orthopaedic regenerative medicine applications.MethodsA systematic search will be performed using PubMed, Elsevier, ScienceDirect, and Google Scholar databases for articles published in English before May 2022. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement and guidelines will be used. Studies will be eligible if they apply to acute and chronic orthopaedic musculoskeletal complications or animal or human disease models. Publications must include the use of MSCs and/or tissue obtained from amniotic/chorionic membrane, amniotic fluid, umbilical cord, and/or umbilical cord-derived Wharton’s jelly as an intervention. Placebos, noninjury models, acute injury models, non-injury models, and gold standard treatments will be compared. The study selection will be performed by two independent reviewers using a dedicated reference management software. Data synthesis and meta-analysis will be performed separately for preclinical and clinical studies.DiscussionThe results will be published in relevant peer-reviewed scientific journals. Investigators will present results at national or international conferences.Trial registration: The Protocol will be registered on PROSPERO international prospective register of systematic reviews prior to commencement.

Nebulized exosomes derived from allogenic adipose tissue mesenchymal stromal cells in patients with severe COVID-19: a pilot study

BackgroundExisting clinical studies supported the potential efficacy of mesenchymal stromal cells as well as derived exosomes in the treatment of COVID-19. We aimed to explore the safety and efficiency of aerosol inhalation of the exosomes derived from human adipose-derived MSCs (haMSC-Exos) in patients with COVID-19.MethodsThe MEXCOVID trial is a phase 2a single-arm, open-labelled, interventional trial and patients were enrolled in Jinyintan Hospital, Wuhan, China. Eligible 7 patients were assigned to receive the daily dose of haMSCs-Exos (2.0 × 108 nano vesicles) for consecutively 5 days. The primary outcomes included the incidence of prespecified inhalation-associated events and serious adverse events. We also observed the demographic data, clinical characteristics, laboratory results including lymphocyte count, levels of D-dimer and IL-6 as well as chest imaging.ResultsSeven severe COVID-19 related pneumonia patients (4 males and 3 females) were enrolled and received nebulized haMSC-Exos. The median age was 57 year (interquartile range (IQR), 43 year to 70 year). The median time from onset of symptoms to hospital admission and administration of nebulized haMSC-Exos was 30 days (IQR, 15 days to 40 days) and 54 d (IQR, 34 d to 69 d), respectively. All COVID-19 patients tolerated the haMSC-Exos nebulization well, with no evidence of prespecified adverse events or clinical instability during the nebulization or during the immediate post-nebulization period. All patients presented a slight increase of serum lymphocyte counts (median as 1.61 × 109/L vs. 1.78 × 109/L). Different degrees of resolution of pulmonary lesions after aerosol inhalation of haMSC-Exos were observed among all patients, more obviously in 4 of 7 patients.ConclusionsOur trial shows that a consecutive 5 days inhalation dose of clinical grade haMSC-Exos up to a total amount of 2.0 × 109 nano vesicles was feasible and well tolerated in seven COVID-19 patients, with no evidence of prespecified adverse events, immediate clinical instability, or dose-relevant toxicity at any of the doses tested. This safety profile is seemingly followed by CT imaging improvement within 7 days. Further trials will have to confirm the long-term safety or efficacy in larger population.Trial Registration: MEXCOVID, NCT04276987.