To investigate the modifying effect of graphene oxide (GO) on small intestinal submucosa (SIS), GO-SIS biocomposite films were fabricated via a non-covalent coating strategy. Structural analyses confirmed the non-covalent interactions between GO and SIS collagen fibers and the preservation of SIS's native fibrous structure. The GO-SIS biocomposite film showed significantly improved hydrophilicity (contact angle: 71.3 ± 1.0°, p < 0.001; water absorption: 159.00 ± 5.60%, p < 0.01) compared to the SIS film. It also exhibited superior mechanical properties under both dry and wet conditions, with significantly higher tensile strength (dry: 24.46 ± 0.99 MPa; wet: 10.16 ± 0.37 MPa) and elongation at break (dry: 11.41 ± 0.55%; wet: 21.26 ± 0.65%) than the SIS film (p < 0.001 for all comparisons). After in vitro degradation, the GO-SIS biocomposite film showed better morphological stability compared to the SIS film. At 4, 8, 16, 24, 48, and 72 hours, the in vitro degradation of the GO-SIS biocomposite film was significantly slower than that of the SIS film. Furthermore, at 4, 8, 16, and 24 hours, the tensile strength and elongation at break of the degraded GO-SIS biocomposite film were significantly higher than those of the degraded SIS film. Biocompatibility assessment indicated no impact on L929 fibroblast viability or proliferation, along with favorable hemocompatibility. Collectively, the non-covalent incorporation of GO effectively enhances the hydrophilicity and mechanical performance of the GO-SIS biocomposite film and slows down its in vitro degradation, offering a promising strategy for the design of advanced tissue repair materials.

Objective: Conventional therapeutic approaches frequently fail to achieve optimal healing and scar reduction in the management of certain extensive and deep wounds. Acellular porcine urinary bladder matrix (UBM) has demonstrated favourable clinical outcomes in tissue regeneration. This study aims to investigate the therapy advantages of combined UBM powder and sheet, and elucidate its whole healing process in comparison to a collagen sponge, using full-thickness skin defect porcine model. Method: Using a full-thickness skin defect porcine model, the porcine urinary bladder was decellularised and lyophilised, then characterised by mass spectrometry for composition analysis. In this model, 4.0 & times;4.0cm full-thickness dorsal wounds were created and randomly divided into three treatment groups: UBM powder and sheet; collagen sponge (Pelnac; Gunze Limited, Inc., Japan); and paraffin gauze (Zhende Medical, China). Wound healing processes were evaluated at two, four and eight weeks post-surgery by gross observation and histopathological staining, as well as immunohistochemical staining of Ki-67, CD31, collagen IV and laminin to assess their expression levels. Tissue regeneration and remodelling were evaluated based on nascent collagen structure and scar degree at eight weeks post-surgery Results: The acellular UBM biological material contained an abundance of protein compositions, which are conducive to tissue regeneration. Combined therapy of UBM improved the healing rate and quality, associated with greater expression of Ki-67, CD31, collagen IV and laminin at two weeks post-surgery. Remodelling results demonstrated superior collagen weave structure and smaller scar area using UBM. Conclusion: In this porcine model, combined therapy of UBM powder and sheet improved deep skin wounds healing and remodelling, making it a promising option for the treatment of deep wounds.

Established quantitative standards for assessing decellularization of biologic scaffolds based on residual DNA levels have been well-documented and widely acknowledged. However, post-implantation complications, such as fever and seroma, are commonly observed which negatively impact clinical outcomes. The presence of cellular debris following decellularization or using source tissues that are naturally high in endotoxin may contribute to the host response to a biologic scaffold. In the study, several multi-step decellularization methods were used to decellularize small intestinal submucosa (SIS) to obtain materials with three distinct levels of residual DNA, lipid residues, and endogenous endotoxin. The potential influence of these residual components on macrophage and lymphocyte polarization in vitro, as well as on the host inflammatory response in vivo post intra-abdominal implantation or abdominal wall defect repair in rats, was assessed. Urinary bladder matrix (UBM) meeting established decellularization criteria and naturally devoid of endotoxin was utilized as a control. The presence of endogenous endotoxin in SIS-ECM resulted in notable changes in macrophage phenotype. SIS-ECM samples with endotoxin levels below FDA limits still upregulated pro-inflammatory factors in vitro. Conversely, SIS with minimal endotoxin content and UBM controls prompted a shift towards a pro-remodeling M2 phenotype, fostering constructive tissue remodeling in a rodent model of abdominal wall defects, irrespective of DNA content. These findings suggest that endotoxin may be a crucial factor influencing biologic scaffolds that are not fully accounted by current decellularization standards. Statement of significance Clinically utilized decellularized biologic scaffolds that meet the established quantitative standards still suffer problems in high incidence of inflammatory complications, including fever and seroma. In this study, we confirmed that endotoxin, rather than residual DNA, is the crucial factor influencing host responses and regenerative outcomes. Tissue sources and decellularization processes are critical for reducing endotoxin levels and attenuating immuno-inflammatory complications. These findings enhance the evaluation of ECM scaffold performance for clinical application, thereby facilitating improved preparation and utilization for tissue defect repairs.

Decellularized biomaterial-based dural patches are clinically employed as dura mater substitutes, owing to their biocompatibility, biodegradability, mechanical flexibility, and efficacy in promoting tissue regeneration. However, these biological grafts have challenges related to patch durability, cerebrospinal fluid leakage, and brain tissue adhesion. In this study, we developed a composite dural patch with a biomimetic structure by encapsulating multilayered small intestinal submucosa (SIS) membranes using basement membrane-containing urinary bladder matrix (UBM) membranes as outer layers, to enhance dural repair. The newly designed UBM@SIS patch demonstrates superior mechanical properties, excellent tissue regeneration capabilities, and enhanced anti-adhesion effects. Immunofluorescent staining revealed a densely organized cell layer anchored to the surface of the UBM@SIS patch, identified as mesothelial cells (Pan-Cytokeratin+/E-cadherin+) and arachnoid barrier cells (ABCs), which are essential for lubricating brain tissue, preventing adhesion, and reducing cerebrospinal fluid leakage. In contrast, only a sparse presence of Pan-Cytokeratin-labeled mesothelial cells and E-cadherin-labeled ABCs was observed on the SIS patch, exhibiting incomplete pia mater and significant adhesion on interface between patch and brain tissue. Furthermore, neurotoxicity evaluations confirmed that the encapsulation of UBM reduces astrogliosis and neuroinflammatory response in early stage. These results suggest that the novel UBM@SIS patch effectively addresses the limitations of existing biological patches and holds significant potential for clinical application.

Natural resorbable collagen membranes are widely used in guided bone regeneration (GBR) in oral implantology and prosthodontics. However, the rapid degradation and inadequate mechanical properties raise concerns about potentially compromising the ultimate bone regeneration efficacy. In this study, we developed a novel GBR membrane by coating a urinary bladder matrix (UBM) onto small intestinal submucosa (SIS), a commonly used acellular material, to improve the cell-barrier and bone-regeneration performances. The results showed that the UBM-SIS membrane exhibited superior tensile strength, high compliance, and slower degradation rate compared to a commercial Bio-Gide membrane, thereby enhancing the support and maintenance properties. In vitro studies indicated enhanced osteogenic behavior and higher osteogenic cytokine expression in human bone marrow-derived mesenchymal stromal cells (hBMSCs) cultured with UBM-SIS extract. In canine mandibular defect models, we proved that the UBM layer effectively resisted fibroblast invasion in the early stage, thereby enhancing the bone meal duration and osteoblast growth. Microcomputed tomography analysis revealed the greater quantity and maturity of bone trabeculae using a UBM-SIS membrane, exhibiting enhanced bone formation at 12 weeks and trabecular maturity at 24 weeks. In conclusion, we proposed a modified resorbable GBR membrane that may improve clinical outcomes in prosthodontics.

Biological scaffolds are widely utilized in hernia treatment due to their exceptional pro-regenerative properties, which mitigate scar formation. However, serious complications occurred, caused by inflammatory response, premature degradation, and mechanical failure. Consequently, improvements of the biological scaffold are necessary to mitigate these risks. In this study, a novel biological scaffold integrating basement membrane-containing urinary bladder matrix (UBM) and small intestinal submucosa (SIS) is developed, and its safety and effectiveness are assessed in comparison to a commercial SIS (c-SIS) scaffold. The introduction of UBM as top surface layers significantly promotes cell adhesion, facilitating rapid formation of isolated regeneration zone. Proteomic analysis has demonstrated a more efficient decellularization of the UBM/SIS scaffold, which subsequently mitigates inflammation in murine models, and promotes the polarization of macrophages toward the pro-healing M2 phenotype in a rat model of abdominal wall muscle defect. Furthermore, a two-year repair trial is conducted on a full-thickness abdominal wall muscle defect in canine model and confirmed that the UBM/SIS scaffold exhibits reduced seroma occurrences and enhanced tissue repair performances. Overall, the efficacy of this novel biological scaffold suggests its potential to minimize hernia recurrence in clinical practice and mitigate patient suffering from severe inflammatory responses.

Objectives:To investigate the clinical value of a novel non-crosslinked biological mesh in laparoscopic inguinal hernia repair.Methods:The prospective randomized controlled study was conducted. The clinical data of 50 adult patients with unilateral inguinal hernia who were admitted to 3 medical centers, including Ruijin Hospital of Shanghai Jiaotong University School of Medicine et al, from September 2019 to March 2020 were selected. Based on random number table, patients were divided into two groups. Patients using the novel non-crosslinked biological mesh in repair surgery were divided into the experiment group and patients using the lightweight, micro-porous, partially absorbable synthetic mesh in repair surgery were divided into the control group. Observation indicators: (1) grouping situations of the enrolled patients; (2) endpoint of the study. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M(range), and comparison between groups was conducted using the non-parameter rank sum test. Count data were described as absolute numbers and (or) persentages, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the non-parameter rank sum test. Repeated measurement data were analyzed using the repeated ANOVA. Taking the recurrence rate of hernia as the basis of efficacy evaluation, according to the intention-to-treat analysis, the confidence interval method (Newcombe Wilson method) was used to conduct non-inferiority statistical analysis of the recurrence rate of hernia between the experiment group and the control group. If the upper limit of 95% confidence interval of the difference of recurrence rate of hernia between the experiment group and the control group is less than 10%, the experiment group is considered to be non-inferior to the control group. Results:(1) Grouping situations of the enrolled patients. A total of 50 adult patients with inguinal hernia were selected for eligibility. There were 44 males and 6 females, aged (60±15)years. All 50 patients were randomly divided into to the experiment group and the control group with 25 cases each. One patient in the control group was not followed up at postoperative month 2, and the rest of 49 patients completed all expected follow-up. No patient in the two groups fell off or were removed. (2) Endpoint of the study. ① The primary endpoint of study. The recurrence rate of hernia was 0 in the experiment group, versus 4%(1/25) in the control group, respectively, showing no significant difference between the two groups ( P>0.05). Results of non-inferiority statistical analysis showed that the 95% confidence interval of the difference of recurrence rate of hernia between the two groups was -19.54% to 9.72%, with the upper limit as 9.72%, which was less than 10%. ② The secondary endpoint of study. There were 2 patients in the control group occurred seroma at postoperative day 14, and none of the rest of patient in the two groups occurred seroma during the follow-up, showing no significant difference in the occurrence of seroma between the two groups ( P>0.05). There was 1 patient in the control group feeling discomfort or foreign body sensation in groin area at postoperative month 2, and none of the rest of patient in the two groups feeling discomfort or foreign body sensation in groin area during the follow-up, showing no significant difference in the feeling discomfort or foreign body sensation in groin area between the two groups ( P>0.05). There was no patient occurred surgical site infection in the experiment group, and there was 1 patient in the control group occurred postoperative skin infection, which had no relationship with mesh. There was no patient in both two groups occurred fever, anaphylaxis and patch related serious adverse reaction during the follow-up. The resting visual analogue scale score, active visual analogue scale score of patients at postoperative 2 days and postoperative 18 months were 0.44±1.00, 1.28±1.46 and 0, 0 in the experiment group, versus 0.40±0.76, 1.28±1.14 and 0.24±1.20, 0.44±1.29 in the control group, respectively. There was a significant difference in the time effect of postoperative active visual analogue scale score of patients between the two groups ( Ftime=10.19, P<0.05). The thickness of the novel non-crosslinked biological mesh before implantation was 0.5?0.7 mm. Two months after operation, results of B-ultrasonic examination in groin area of 10 patients from the experiment group showed a strong echo area at the patch implant area with a thickness as 2 mm. Conclusion:Application of novel non-crosslinked biological mesh in laparoscopic inguinal hernia repair is safe and effective.

Introduction and hypothesis To investigate the tissue reactions of a novel porcine-derived urinary bladder matrix/small intestinal submucosa (UBM/SIS) biological mesh and SIS mesh implanted in a rabbit vaginal defect model. Methods Thirty-two rabbits were implanted with UBM/SIS mesh (Group A) and SIS mesh (Group B), respectively. Rabbits were sacrificed at 7, 14, 60, and 180 days after implantation. The tensile strength, elongation at break, and elastic modulus of the tissue were measured using biomechanical methods. The inflammatory response, cell infiltration, vascularization, and collagen fibers were observed. Results Compared with Group B, the tensile strength and elongation at break of group A was higher at 14, 60, and 180 days. The elastic modulus of group A was lower at 180 days. Inflammatory response of group A was milder at 14, 60, and 180 days. There was more cell infiltration in group A at 7 and 14 days. Vascularization was higher in group A at 7 days and 14 days. The order of collagen in group A was better at 14, 60, and 180 days. The proportion of thick red fibers in both groups showed an increasing trend. At 14 days, group A had more thick red fibers. Conclusions The novel UBM/SIS composite mesh had a milder inflammatory response; earlier induction of cell infiltration, angiogenesis, and collagen regeneration. Collagen fibers had a better order. It has a higher tensile strength and greater elongation at break, and can be used as a potential material for the treatment of pelvic organ prolapse.

目的 比较3种不同组织来源生物补片的体内外降解性能.方法 采用Ⅰ型胶原酶对来源于基底膜(BM)与小肠黏膜下层(SIS)复合细胞外基质、单纯小肠黏膜下层、戊二醛化学交联心包(PC)三种生物补片进行体外降解试验,确定各材料的降解率;采用各材料修补大鼠腹壁肌部分层次缺损评估3种补片的降解和组织修复情况.结果 体外降解试验中,酶溶液作用120 h后,非交联的BM/SIS复合补片和SIS补片完全降解,PC补片的降解率仅为4.3％±1.9％.体内降解试验中,术后2个月,大体观和组织病理切片染色结果证实BM/SIS复合补片完全降解,组织重塑,再生胶原有序;SIS补片完全降解,再生胶原有序性较差;术后12个月,PC补片未见明显降解,未见细胞长入补片中央区.结论 虽然BM/SIS复合补片的体内外完全降解时间均较短,但其可以实现组织的良好快速重塑,提示BM/SIS复合补片是一种降解与再生同步的组织修复材料.

目的 筛选合适的抗菌剂制备生物抗菌修补片,验证抑菌效果和安全性.方法 评估硝酸银、苯扎溴铵和三氯生的体外浮游、生物膜状态革兰氏阴/阳性菌的抑菌效果,选择合适浓度的抗菌剂与猪小肠黏膜下层脱细胞基质结合,制备生物抗菌修补片,考察补片的体外抑菌性,抗菌剂释放和细胞毒性.结果 硝酸银对革兰氏阴性菌、苯扎溴铵对革兰氏阳性菌抑菌效果较好,但对生物膜状态的细菌抑制作用较差.三氯生对浮游菌和细菌生物膜均具有良好抑菌效果.故选择三氯生和猪小肠黏膜下层制备生物抗菌修补片(TSIS).体外释放实验证实TSIS可以持续释放三氯生,对金葡菌体外抑菌作用长达24 d、大肠杆菌的抑菌作用长达7 d.细胞毒性实验证实TSIS的生物安全性良好.结论 新型生物抗菌修补片可持续释放抗菌成分,生物安全性良好.

Objective To evaluate the therapeutic effects of biological grafts derived from different tissue,to provide reference information for the clinical choice of biological grafts.Methods Healthy SD rats were randomly divided into 5 groups (n=10).Bilateral partial thickness defect in abdominal wall of rats were created and repaired with either basement membrane (BM)/small intestine submucosa (SIS)composite extracellular matrix (ECM) graft,SIS,dermis or pericardium,while untreated defects were served as control.Animals were sacrificed at 2,4,8 and 16 weeks after surgery,the incidence of seroma,shrinkage in repair area and degradation of implants were recorded.The repaired abdominal walls were harvested for histological evaluation to observe cell ingrowth,neovascularization,and fibrous encapsulation.Results No seroma formation was observed in BM/SIS composite ECM graft repaired samples,and the samples replaced with dense and well-organized collagen fibers with mainly initial dimensions at 4 weeks post-surgery,degraded at 8 weeks post-surgery.There still presents massive inflammatory cells infiltration in SIS repaired area at 4 weeks post-surgery.At 8 weeks post-surgery,seroma incidence in SIS repaired samples was 65％ and SIS was degraded.SIS had reconstructed tissue defects with a notable shrinkage rate of-52.0％±9.8％.50％ of dermis repaired area was infiltrated by cells and fully degraded.Fibrous encapsulation was formed in other dermis repaired area and cells were only infiltrated in the interface area.Significant enlargement compared with original implant area and no obvious degradation were observed.During the experimental period,scarcely any cells infiltrated the scaffold of pericardium with a shrinkage rate of-29.5％±14.0％ at 16 weeks post-surgery.No degradation was observed and fibrous encapsulations were formed in pericardium repaired samples.Conclusion BM composite graft has shown a better tissue regeneration compared with SIS,dermis and pericardium.

Objective To explore the mechanical properties of the hybrid patch made of porcine acellular small intestinal submucosa (P-SIS) and polydioxanone (PDO) and its effect on repairing the abdominal wall defects of rats.Methods The PDOSIS patch and 8SIS patch were constructed by weaving and vacuum lamination and the mechanical properties were measured.In the both sides of abdomen of 36 healthy female SD rats,a partially defect (lcm × lcm) was created by surgery and repaired with a same area of PDO-SIS patch or 8SIS patch,each 12 rats were randomly sacrificed at 2,4 and 8 weeks after surgery,respectively,and tissue regeneration was evaluated.Results The bending length and tensile strength of the PDO-SIS patch were stronger than the 8SIS patch (P＜0.05) with the increase of diameter.No acute inflammation occurred at the repaired sites of the two groups,but a moderate chronic inflammation was observed 2 weeks after surgery,and the inflammatory response reduced gradually over time,no obvious chronic inflammation was found in the 8th week after surgery,with no statistical differences between the two groups (P＞0.05).Two patch materials were degraded gradually in the repaired area,and the regenerated collagen tissues were deposited continuously,but no significant differences existed between the two groups in the collagen content and collagen arrangement (P＞0.05).Conclusion The PDO-SIS patch has better mechanical properties than the 8SIS patch,and does not cause strong immune rejection when used to repair abdominal partially defect of SD rats.

In this dataset, we particularly depicted the harvest and perfusion decellularization of porcine rectus abdominis (RA), accompanied with displaying of the retained vascular trees within the perfusion-decellularized skeletal muscle matrix (pM-ECM) using vascular corrosion casting. In addition, several important tips for successful pM-ECM preparation were emphasized, which including using anatomically isolated skeletal muscle as tissue source with all main feeding and draining vessels perfused, preserving the internal microcirculation availability, aseptic technique and pyrogen free in all steps, sequential perfusion via artery or vein, and longtime washing after decellularization. The data are supplemental to our original research article describing detailed associations of pM-ECM as a clinically relevant scale, three-dimensional scaffold with a vascular network template for tissue-specific regeneration, “Perfusion-decellularized skeletal muscle as a three-dimensional scaffold with a vascular network template” Zhang et al. (2016) [1].

There exists a great need for repair grafts with similar volume to human skeletal muscle that can promote the innate ability of muscle to regenerate following volumetric muscle loss. Perfusion decellularization is an attractive technique for extracellular matrix (ECM) scaffold from intact mammalian organ or tissue which has been successfully used in tissue reconstruction. The perfusion-decellularization of skeletal muscle has been poorly assessed and characterized, but the bioactivity and functional capacity of the obtained perfusion skeletal muscle ECM (pM-ECM) to remodel in vivo is unknown. In the present study, pM-ECM was prepared from porcine rectus abdominis (RA). Perfusion-decellularization of porcine RA effectively removed cellular and nuclear material while retaining the intricate three-dimensional microarchitecture and vasculature networks of the native RA, and many of the bioactive ECM components and mechanical properties. In vivo, partial-thickness abdominal wall defects in rats repaired with pM-ECM showed improved neovascularization, myogenesis and functional recellularization compared to porcine-derived small intestinal submucosa (SIS). These findings show the biologic potential of RA pM-ECM as a scaffold for supporting site appropriate, tissue reconstruction, and provide a better understanding of the importance maintaining the tissue-specific complex three-dimensional architecture of ECM during decellularization and regeneration.

The invention relates to a canine uterus whole-organ acellular matrix and a preparation method as well as the application of the canine uterus whole-organ acellular matrix. The canine uterus whole-organ acellular matrix is prepared by the following steps: selecting raw materials, pre-treating, removing cells, sterilizing and storing. The canine uterus whole-organ acellular matrix can be used for preparing particles, fluid compositions, gels and active peptides. The canine uterus whole-organ acellular matrix completely reserves the complex three-dimensional array of the natural uterus extracellular matrix and the horizontal-inside longitudinal-outside ultra-structural of the smooth muscle membrane; the canine uterus whole-organ acellular matrix also reserves the complete and non-leaking vascular matrix network as well as reserves a large number of biological active ingredients such as growth factors, hyaluronic acid, amino glucan and laminin, so that the canine uterus whole-organ acellular matrix has high affinity to the endometrial stem cells. Thus, the canine uterus whole-organ acellular matrix has the biomechanical strength and tenacity similar to those of the natural uterus; moreover, the canine uterus whole-organ acellular matrix is thorough in cell removal, free of obvious immune rejection and free of ethical and moral issues.

Are non-crosslinked biologic mesh effective for laparoscopic inguinal hernia repair: Results of a multicenter, non-randomized controlled study Jian Zhang, Gan Yao, Zhi-chao Tian, Wei-xing Yu, Hai-yang Zhou, Wen-yue Cheng, Rong-jia He, Qiang Wang, Zhi-qian Hu Department of Surgery, Shanghai Chang Zheng Hospital, Second Military Medical University, China, Department of Regenerative Medicine, Shanghai Zhabei District Central Hospital, China Department of General Surgery, Foshan First People's Hospital, China, Department of General Surgery, Shanghai Zhabei District Central Hospital, China Department of Urology, Shangyu People's Hospital, China, Department of General Surgery, Zhongshan People's Hospital, China