异种脱细胞膀胱黏膜下层支架的制备

Preparation of xenogeneic acellular urinary bladder submucosa scaffold

目的构建一种具有良好组织相容性的脱细胞膀胱黏膜下层支架,探讨其作为生物材料的可行性。方法取健康猪膀胱依次置于不同浓度的叠氮钠溶液中。采用低渗处理、液氮反复冻融以及NaOH溶解等方法对猪膀胱进行脱细胞处理。通过组织学检测和紫外分光光度法确定脱细胞效率;细胞毒性试验、细胞黏附实验、以及皮下植入试验评价异种脱细胞膀胱黏膜下层支架的生物安全性。结果组织学观察结果显示,经脱细胞处理后支架内无细胞残留;与对照组比较,支架内残留的DNA含量显著降低。细胞毒性实验结果显示,支架浸提液对人膀胱平滑肌细胞的细胞活力无显著影响。细胞黏附实验结果显示,人膀胱平滑肌细胞能够在支架上贴附生长。皮下植入试验结果显示,支架植入后,炎症反应轻微,术后4周未见明显的炎性细胞浸润。结论经脱细胞处理获得的膀胱黏膜下支架具有良好的生物安全性,是一种安全的可植入材料。

Objective To construct the acellular urinary bladder submucosa scaffold with good histocompatibility and to further explore its possibility as a promising biomaterial infurture.Methods The bladder of healthy pigs was placed in different concentrations of sodium azide solution insequence.Then the porcine bladder was decellularized using methods such as hypotonic solution,freeze-thawed treatment with liquid nitrogen and NaOH spallation,respectively.Firstly,the acellul are fficiency was identified by both histological examination and ultraviolet spectrophotometry.Next,the biological assessment of xenogeneic acellular urinary bladder submucosa scaffold was evaluated by cytotoxicity assay,cell adhesion assay,as well as subcutaneous implant experiment.Results The observation from histology showed no cell residue in this scaffold after acellular treatment.Also,the residual DNA content in scaffold decreased significantly compared with control group.Meanwhile,cytotoxicity assay showed the extracts of scaffold had no significant effect on the cell viability of human bladder smooth muscle cells.Cell adhesion assay suggested that this scaffold represented an adhesion ability for human smooth muscle cells.Moreover,the subcutaneous implant experiment confirmed that the inflammatory reaction was markedly mild after implantation,and there was no significant inflammatory infiltration at 4 weeks after operation.Conclusion This urinary bladder submucosa scaffold obtained by acellular treatment exhibits a good biocompatibility,which is a safe and promising implant material.