生物素化去细胞猪主动脉瓣构建组织工程心脏瓣膜研究

Biotinylated porcine decellularized aortic valve enhances attachment of mesenchymal stem cells in tissue engineered heart valve

目的制备生物素化去细胞猪主动脉瓣，通过生物素-亲和素系统提高去细胞瓣膜的细胞黏附效率。方法将10mmol／L的生物素和5g／L亲和素分别依次与去细胞猪主动脉瓣反应，制备表面含亲和素的生物素化去细胞猪主动脉瓣。将1．0×10^3生物素化和未生物素化的骨髓基质干细胞，分别种植于上述改性前后的去细胞瓣膜，应用Hoechst 33258试验确定细胞的黏附效率，扫描电镜观察细胞在瓣膜上的生长。结果生物素和亲和素能链接到去细胞猪主动脉瓣，生物素化骨髓基质干细胞在表面含亲和素的生物素化去细胞猪主动脉瓣上的黏附效率（55．73±4．53）％显著高于各对照组分别为（31．20±3．94）％、（30．75±3．15）％和（28．51±4．52）％，且扫描电镜示细胞在去细胞猪主动脉瓣膜表面形成-细胞层，生长良好。结论去细胞猪主动脉瓣可生物素化，并能以亲和素为中介，与细胞表面生物素的结合，增加细胞的黏附效率，且不影响细胞的生长。

Objective To evaluate the utilization of biotinylated porcine decellularized aortic valve （B-PDAV） to improve cell attachment. Methods Porcine aortic valves were decellularized using detergent. Ten mmol/L Biotin and 5 g/L avidin were introduced into the decellularized valves sequentially. The modified scaffolds were seeded with 1.0 × 10^5 biotinylated mesenchymal stem cells （B-MSC）. As controls, PDAV was seeded with B-MSC, PDAV was seeded with MSC, and B-PDAV was seeded with MSC in the same manner. Hoechst 33258 assay was performed to determine the efficiency of cell attachment in each group. Cell growth on the scaffold was evaluated under the scanning electron microscopy. Resuits PDAV could be biotinylated and further conjugated with avidim The efficiency of cell attachment in the experimental and the control groups was （ 55.73 ± 4.53 ） % , （ 31.20 ±3.94 ） % , （ 30.75 ±3.15 ） % and （28.51± 4.52）% respectively. The difference in cell attachment between the experimental group and the control group was significant. Scanning electron microscopy revealed that cells adhered and grew well on this scaffold. Conclusion B-PDAV could enhance the attachment of B-MSC through the connection of avidin.