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Reproduction of different tissues using scaffolds and materials is a major element in regenerative medicine. The regeneration of whole organs with decellularized extracellular matrix (dECM) has remained a goal despite the use of these materials for different purposes. Recently, decellularization techniques have been widely used in producing scaffolds that are appropriate for regenerating damaged organs and may be able to overcome the shortage of donor organs. Decellularized ECM offers several advantages over synthetic compounds, including the preserved natural microenvironment features. Different decellularization methods have been developed, each of which is appropriate for removing cells from specific tissues under certain conditions. A variety of methods have been advanced for evaluating the decellularization process in terms of cell removal efficiency, tissue ultrastructure preservation, toxicity, biocompatibility, biodegradability, and mechanical resistance in order to enhance the efficacy of decellularization methods. Modification techniques improve the characteristics of decellularized scaffolds, making them available for the regeneration of damaged tissues. Moreover, modification of scaffolds makes them appropriate options for drug delivery, disease modeling, and improving stem cells growth and proliferation. However, considering different challenges in the way of decellularization methods and application of decellularized scaffolds, this field is constantly developing and progressively moving forward. This review has outlined recent decellularization and sterilization strategies, evaluation tests for efficient decellularization, materials processing, application, and challenges and future outlooks of decellularization in regenerative medicine and tissue engineering.

We aimed to investigate the independent outcome predictors of continuous antibiotic prophylaxis (CAP) in vesicoureteral reflux, train a model to predict the outcome, and evaluate which infants should be referred for endoscopic vesicoureteral reflux correction in their first visits. A total of 225 infants ≤ 2 years of age with a diagnosis of vesicoureteral reflux between 2009 and 2022 were recruited; 115 patients from a pediatric nephrology clinic received CAP, and 110 patients from a pediatric urology department underwent endoscopic injection of dextranomer/hyaluronic acid copolymer. In the multivariable analysis, only renal scarring and bladder dysfunction were significantly associated with post-treatment febrile urinary tract infections and/or renal scarring and vesicoureteral reflux persistence, respectively, in children who received CAP. The machine learning modeling showed that for both febrile urinary tract infections and/or renal scarring and vesicoureteral reflux persistence, the random forest was the best fit. On the other hand, we observed that the success rates of endoscopic injection among the patients with renal scarring and bladder dysfunction were acceptable. In conclusion, renal scarring and bladder dysfunction were predictors of vesicoureteral reflux outcomes when the infant was receiving CAP. Therefore, referring these patients to a urologist is advised during their first visits as they benefit from endoscopic injection.

Despite recent advancements in peripheral nerve regeneration, the creation of nerve conduits with chemical and physical cues to enhance glial cell function and support axonal growth remains challenging. This study aimed to assess the impact of electrical stimulation (ES) using a conductive nerve conduit on sciatic nerve regeneration in a rat model with transection injury. The study involved the fabrication of conductive nerve conduits using silk fibroin and Au nanoparticles (AuNPs). Collagen hydrogel loaded with green fluorescent protein (GFP)-positive adipose-derived mesenchymal stem cells (ADSCs) served as the filling for the conduit. Both conductive and non-conductive conduits were applied with and without ES in rat models. Locomotor recovery was assessed using walking track analysis. Histological evaluations were performed using H&E, luxol fast blue staining and immunohistochemistry. Moreover, TEM analysis was conducted to distinguish various ultrastructural aspects of sciatic tissue. In the ES + conductive conduit group, higher S100 ( p  < 0.0001) and neurofilament ( p  < 0.001) expression was seen after 6 weeks. Ultrastructural evaluations showed that conductive scaffolds with ES minimized Wallerian degeneration. Furthermore, the conductive conduit with ES group demonstrated significantly increased myelin sheet thickness and decreased G. ratio compared to the autograft. Immunofluorescent images confirmed the presence of GFP-positive ADSCs by the 6th week. Locomotor recovery assessments revealed improved function in the conductive conduit with ES group compared to the control group and groups without ES. These results show that a Silk/AuNPs conduit filled with ADSC-seeded collagen hydrogel can function as a nerve conduit, aiding in the restoration of substantial gaps in the sciatic nerve with ES. Histological and locomotor evaluations indicated that ES had a greater impact on functional recovery compared to using a conductive conduit alone, although the use of conductive conduits did enhance the effects of ES.

To evaluate the effectiveness of the Complete Primary Repair of Exstrophy (CPRE) technique for bladder exstrophy-epispadias complex (BEEC) reconstruction and its comparison with the Modern Staged Repair of Exstrophy (MSRE) technique. A comprehensive literature review of CPRE and MSRE was conducted, focusing on factors such as continence rates, postoperative outcomes, and complications. Various studies on pelvic biometry, surgical approaches, and long-term evaluations of renal function and continence were analyzed. CPRE demonstrates promising outcomes, particularly in resource-limited settings, by reducing the number of surgeries and associated risks compared to MSRE. Success rates for continence post-CPRE were higher when performed correctly in the first attempt, with fewer complications such as vesicoureteral reflux and hydronephrosis. Pelvic biometry assessments, including bladder neck placement and levator ani angle, were significant predictors of successful continence outcomes. CPRE offers a viable alternative to MSRE, particularly in low-resource environments, with favorable continence and renal outcomes. Proper surgical execution in the first attempt is critical for long-term success, emphasizing the importance of surgeon expertise and postoperative care. Further long-term studies are necessary to solidify CPRE's role as the primary surgical approach for BEEC.

Purpose Successful in vitro transplantation of spermatogonial stem cells (SSCs) demands effective culture systems for SSCs proliferation and differentiation. Natural extracellular matrix (ECM) creates a microenvironment suitable for culture of stem cells. In the present study, we intended to assess the capability of the porous scaffold consisting of hyaluronic acid (HA), chitosan, and decellularized testicular matrix (DTM) as a proper niche for SSCs seeding. Methods The testes of four NMRI mice were extracted for further detergent-based decellularization process. We isolated, cultured, and clarified neonate mouse SSC, and a three-dimensional scaffold was prepared for SSCs culture. The loaded SSCs and hydrogel-based scaffold were investigated by several studies including scanning electron microscopy (SEM), 4′,6-diamidino-2-phenylindole (DAPI), 3-[4, 5-dimethyl (thiazol-2yl)-3,5diphenyl] tetrazolium bromide (MTT), Acridine orange, and Immunohistochemistry (IHC) staining. Results The efficiency of decellularization process was confirmed by DAPI, hematoxylin and eosin (H&E), and Masson’s Trichrome staining. Acridine orange also depicted SSCs proliferation and viability. SEM approved the preservation of ECM components and also showed complex, coiled, and tubular seminiferous tubules, with intact and condensed collagenous form of the tunica albuginea. MTT test also revealed the scaffold’s non-toxicity. Expression of PLZF, TP1, and TEKT1 markers also verified the capacity of SSCs proliferation on a cogel scaffold. Conclusion In conclusion, cogel scaffold consisting of DTM, HA, and chitosan may provide the supporting layer for in vitro SSC differentiation and proliferation.

Chronic kidney disease poses a significant threat to public health. Renal replacement therapy is the primary treatment option for end-stage kidney disease. However, there is a promising and relatively new method in regenerative medicine for creating a functional organ known as whole kidney decellularization. This method uses the intrinsic vasculature to perfuse the decellularizing agent into the tissue, effectively penetrating and removing cellular material. The regenerated bioscaffolds could serve as a source of organ donation. This study is focused on evaluating the effectiveness of various SDS exposures in decellularizing human fetal kidneys. The study included human fetal kidneys harvested from fetuses terminated before 14 weeks of gestational age. Kidneys were divided into six treatment groups based on SDS concentration and duration of perfusion. Decellularization, scanning electron microscopy, histopathological staining, immunofluorescent staining, and immunohistochemistry staining were performed to evaluate the adequacy of the process. The statistical analysis revealed that the SDS 0.1% treatment group had the highest collagen deposition after 24 h, significantly greater than the SDS 0.5% treatment group at 24 and 48 h. No significant differences were observed among the other treatment groups. The study concludes that the SDS 0.1% treatment group for 24 h was the most effective in terms of ECM content preservation and effective cell removal. This treatment showed better results than the other treatment groups and can be considered for future whole kidney decellularization studies.

Introduction Over the past two decades, there has been a notable rise in the number of individuals afflicted with End-Stage Renal Disease, resulting in an increased demand for renal replacement therapies. While periodic dialysis is beneficial, it can negatively impact a patient’s quality of life and does not fully replicate the secretory functions of the kidneys. Additionally, the scarcity of organ donors and complications associated with organ transplants have underscored the importance of tissue engineering. Regenerative medicine is revolutionized by developing decellularized organs and tissue engineering, which is considered a cutting-edge area of study with enormous potential. Developing bioengineered kidneys using tissue engineering approaches for renal replacement therapy is promising. Method and materials We aimed to systematically review the essential preclinical data to promote the translation of tissue engineering research for kidney repair from the laboratory to clinical practice. A PubMed search strategy was systematically implemented without any linguistic restrictions. The assessment focused on complete circumferential and inlay procedures, thoroughly evaluating parameters such as cell seeding, decellularization techniques, recellularization protocols, and biomaterial types. Results Of the 1,484 studies retrieved from the following primary searches, 105 were included. Kidneys were harvested from eight different species. Nine studies performed kidney decellularization from discarded human kidneys. Sixty-four studies performed whole organ decellularization. Some studies used acellular scaffolds to produce hydrogels, sheets, and solutions. Decellularization is achieved through physical, chemical, or enzymatic treatment or a combination of them. Sterilization of acellular scaffolds was also thoroughly and comparatively evaluated. Lastly, different recellularization protocols and types of cells used for further cell seeding were demonstrated. Conclusion A comprehensive review of the existing literature about kidney tissue engineering was conducted to evaluate its effectiveness in preclinical investigations. Our findings indicate that enhancements in the design of preclinical studies are necessary to facilitate the successful translation of tissue engineering technologies into clinical applications.

Background Retinal degenerative disorders (RDDs) cause vision loss by damaging retinal neurons and photoreceptors, affecting individuals of all ages. Cell-based therapy has emerged as an effective approach for the treatment of RDDs with promising results. This meta-analysis aims to comprehensively evaluate the efficacy of cell therapy in treating age-related macular degeneration (AMD), retinitis pigmentosa (RP), and Stargardt macular degeneration (SMD) as the most prevalent RDDs. Methods PubMed, Scopus, Web of Science, and Embase were searched using keywords related to various retinal diseases and cell therapy treatments until November 25th, 2023. The studies’ quality was evaluated using the Joanna Briggs Institute’s (JBI) checklist for quasi-experimental studies. Visual acuity measured as LogMAR score was used as our main outcome. A three-level random-effect meta-analysis was used to explore the visual acuity in patients who received cell-based therapy. Heterogeneity among the included studies was evaluated using subgroup and sensitivity analyses. Moreover, meta-regression for the type of cells, year of publication, and mean age of participants were performed. Results Overall, 8345 studies were retrieved by the search, and 39 met the eligibility criteria, out of which 18 studies with a total of 224 eyes were included in the meta-analysis. There were 12 studies conducted on AMD, 7 on SMD, and 2 on RP. Cell therapy for AMD showed significant improvement in LogMAR ( p  < 0.05). Also, cell therapy decreased the LogMAR score in SMD and RP ( p  < 0.01 and p  < 0.0001, respectively). Across all conditions, no substantial publication bias was detected (p < 0.05). Conclusion The findings of the study highlight that the application of cell therapy can enhance the visual acuity in AMD, SMD, and RP.

Extracellular matrix (ECM) remodeling in cancer provides an essential substructure for tumor progression. We utilized patient-derived scaffolds (PDS) to model tumor ECM changes and study their impact on cell behavior. PDS were obtained from breast tumor and normal healthy breast tissue by decellularizing surgically resected specimens. We used PDS to design a 3D culture of the breast cancer cell line MCF-7. We utilized bioinformatics pipeline to identify hub genes indicative of cell invasiveness, and assessed their expression using quantitative real-time PCR. Our decellularization protocol led to decellularization of tissues while preserving key ECM components. ECM components such as collagen, glycosaminoglycans, collagen IV, and vimentin were significantly overexpressed in tumor compared to normal PDS. In 3D cultures, cells cultured on normal PDS had significantly lower viability and proliferation. Moreover, cells cultured for 15 days on tumor PDS showed significant overexpression of hub genes, CAV1, CXCR4, CNN3, MYB, and TGFB1, and secreted higher levels of IL-6 (122.91 vs. 30.23 pg/10⁶ cells, P  < 0.05), all markers of an aggressive breast cancer phenotype. Breast cancer cells fail to acquire aggressive features when cultured in an ECM lacking tumor-specific alterations. This underscores the potential of therapeutic approaches targeting the mechanobiological properties of the tumor ECM.

Severely immunocompromised NOD.Cg‐PrkdcscidIl2rgtm1Sug (NOG) mice are among the ideal animal recipients for generation of human cancer models. Transplantation of human solid tumors having abundant tumor‐infiltrating lymphocytes (TILs) can induce xenogeneic graft‐versus‐host disease (xGvHD) following engraftment and expansion of the TILs inside the animal body. Wilms' tumor (WT) has not been recognized as a lymphocyte‐predominant tumor. However, 3 consecutive generations of NOG mice bearing WT patient‐derived xenografts (PDX) xenotransplanted from a single donor showed different degrees of inflammatory symptoms after transplantation before any therapeutic intervention. In the initial generation, dermatitis, auto‐amputation of digits, weight loss, lymphadenopathy, hepatitis, and interstitial pneumonitis were observed. Despite antibiotic treatment, no response was noticed, and thus the animals were prematurely euthanized (day 47 posttransplantation). Laboratory and histopathologic evaluations revealed lymphoid infiltrates positively immunostained with anti‐human CD3 and CD8 antibodies in the xenografts and primary tumor, whereas no microbial infection or lymphoproliferative disorder was found. Mice of the next generation that lived longer (91 days) developed sclerotic skin changes and more severe pneumonitis. Cutaneous symptoms were milder in the last generation. The xenografts of the last 2 generations also contained TILs, and lacked lymphoproliferative transformation. The systemic immunoinflammatory syndrome in the absence of microbial infection and posttransplant lymphoproliferative disorder was suggestive of xGvHD. While there are few reports of xGvHD in severely immunodeficient mice xenotransplanted from lymphodominant tumor xenografts, this report for the first time documented serial xGvHD in consecutive passages of WT PDX‐bearing models and discussed potential solutions to prevent such an undesired complication. Three consecutive generations of NOG mice bearing Wilms tumor patient‐derived xenografts from a single donor showed different degrees of immunoinflammatory symptoms. Histopathologic evaluations of the xenografts and parental tumor showed lymphoid infiltrates and human‐originated CD3 and CD8 positive cells in the tissue blocks, while no lymphoproliferative disorder was evident (suggestive of a lymphodominant tumor graft). In the absence of microbial infection and lymphoproliferative disorder, the syndrome was suggestive of xenogeneic graft‐versus‐host disease.