负载抗sca-1抗体和碱性成纤维细胞生长因子的脱细胞膀胱基质的生物性能研究

Evaluation of biocompatibility of porcine urinary bladder matrix loaded with both anti-sca-1 and basic fibroblast growth factor

目的观察负载抗sca-1抗体(anti-sca-1)和碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)-脱细胞膀胱基质(urinary bladder matrix,UBM)生物支架的生物学性能,探讨其作为新型盆底修复材料的可能性。方法通过双特异性化学交联试剂4-(N-马来酰亚胺甲基)环己烷-1-羧酸磺酸基琥珀酰亚胺酯钠盐(sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate,sulfosmcc)将anti-sca-1和b FGF特异性结合到UBM上,得到anti-sca-1/b FGF-UBM生物支架。通过扫描电子显微镜观察生物支架表面结构,CCK-8(cell counting kit-8)间接检测体外富集sca-1阳性细胞情况,免疫荧光观察支架诱导小鼠骨髓间充质干细胞(mouse mesenchymal stem cells,m MSCs)分化成熟情况。结果扫描电子显微镜显示anti-sca-1和b FGF能特异性的结合到UBM上。anti-sca-1和b FGF组荧光强度明显高于对照组(P<0.05)。anti-sca-1/b FGF-UBM生物支架富集的sca-1阳性细胞明显较对照组多(P<0.05)。免疫荧光观察结果提示anti-sca-1/b FGF-UBM生物支架可以促进m MSCs向平滑肌细胞分化。结论 anti-sca-1/b FGF-UBM生物支架显示出了良好的生物性能,既能富集sca-1阳性细胞,又能进一步促进细胞分化成熟。

Objective To evaluate the biocompatibility of porcine urinary bladder matrix（ UBM）loaded with anti-sca-1 antibody and basic fibroblast growth factor（ b FGF） and test its feasibility as a new material for repairing pelvic prolapse. Methods A bispecific cross-linking reagent sulfosuccinimidyl 4-[Nmaleimidomethyl] cyclohexane-1-carboxylate（ sulfo-smcc） was used to bind anti-sca-1 and b FGF to the UBM to obtain the anti-sca-1/b FGF-UBM biological scaffold. The structure of this scaffold material was observed using scanning electron microscopy,and its performance in enriching sca-1-positive cells in vitro was evaluated using cell counting kit-8（ CCK-8） assay. Immunofluorescence assay was employed to assess the differentiation of mouse mesenchymal stem cells（ m MSCs） seeded in this scaffold material. Results Scanning electron microscopy showed that anti-sca-1 and b FGF could specifically bind to the UBM. The anti-sca-1/b FGF-UBM biological scaffold exhibited a significantly stronger fluorescence intensity than the control material（ P 0. 05）and effectively enriched sca-1-positive cells in vitro（ P 0. 05）. Immunofluorescence assay confirmed that m MSCs seeded in this scaffold material were capable of differentiating into smooth muscle cells. Conclusion The anti-sca-1/b FGF-UBM scaffold shows a good biocompatibility.