不同肌腱处理方法对同种异体肌腱生物学性能的影响

Biological properties after different allogeneic tendon managements

目的探讨不同处理方法对同种异体肌腱生物学性能的影响,并筛选最佳处理方法。方法应用四种处理方法:1%TnBP+1%Triton X-100(A组)、0.5%胰蛋白酶+0.5%Triton X-100(B组)、1%TnBP(C组)、1%Triton X-100+70%异丙醇(D组)分别对新鲜肌腱进行处理,组织学染色观察去细胞效果;CCK8法检测细胞毒性;扫描电镜观察细胞黏附情况;皮内刺激实验评价体内相容性。结果采用0.5%胰蛋白酶+0.5%Triton X-100去除细胞最彻底,纤维排列整齐,结构完整,其它组均观察到有少部分细胞残留;CCK8结果显示,A组和C组处理的肌腱为二级细胞毒性,B组和D组处理的肌腱为1级细胞毒性。扫描电镜结果显示A组和C组处理的肌腱上细胞黏附数量较少,且基本呈圆形;D组处理的肌腱上细胞黏附数量较多,细胞状态较好;B组处理的肌腱上细胞黏附数量最多,细胞成片生长,状态较好。皮内刺激实验结果显示,各组均出现红肿和焦痂。结论在四种肌腱处理方法中,0.5%胰蛋白酶+0.5%Triton X-100的处理方法最为理想。

Objective To investigate different biological properties after four allogeneic tendon managements,and to select the best option.Methods Group division:1%TnBP+1%Triton X-100(Group A);0.5%trypsin+0.5%Triton X-100(Group B);1%TnBP(Group C);1%Triton X-100+70%isopropanol(Group D).Four methods above were applied in the fresh tendon to observe the decellularization after histological staining.CCK8,scanning electron microscopy(SEM),and intradermal stimulation test were used to detect cytotoxicity,observe cell adhesion,and evaluate in vivo compatibility respectively.Results Cells were removed most thoroughly in Group B with neatly arranged fibers and intact structure.Cell residues were observed in other groups.CCK8 results:grade 2 cytotoxicity in Group A and Group C;grade 1 cytotoxicity in Group B and Group D.SEM results:less cell adhesion in Group A and Group C,and most cells were in round-shape;more cell adhesion in Group D,and cells were in a better state;most cell adhesion in Group B,and cells grew in piece of a better state.Intradermal stimulation results:redness,swelling,and eschar in each group.Conclusions 0.5%trypsin+0.5%Triton X-100 is an ideal option in the allogeneic tendon management.