夹带剂对超临界萃取技术处理肌腱的影响

Effect of entrainer on tendon treated by supercritical fluid extraction

[目的]研究不同夹带剂对超临界萃取技术(supercritical CO2 fluid extraction, SCCO2)处理肌腱的影响。[方法]将同种异体肌腱随机分为4组,A组为未经SCCO2处理的同种异体肌腱;B组为以无水乙醇作为夹带剂经SCCO2处理的同种异体肌腱;C组为以无水乙醇+1%乙二胺四乙酸(EDTA)作为夹带剂经SCCO2处理的同种异体肌腱;D组为以(环氧乙烷/环氧丙烷)共聚醚二醇(EO/PO)作为夹带剂经SCCO2处理的同种异体肌腱。对4组肌腱行力学测试、组织学观察和体外细胞增殖毒性试验。[结果] 4组同种异体肌腱的最大载荷差异无统计学意义(P>0.05);与A组相比,D组的抗拉强度变小(P>0.05),且弹性模量显著变小以及断裂伸长率显著增大(P<0.05),而B组和C组上述指标无显著变化(P>0.05)。组织学观察显示,D组肌腱细胞外基质完整且无可见细胞核。体外细胞增殖试验显示经SCCO2处理的同种异体肌腱在B组、C组和D组均未抑制体外细胞增殖。[结论]在添加(环氧乙烷/环氧丙烷)共聚醚二醇作为夹带剂的SCCO2处理方法能获得理想的无细胞毒性脱细胞同种异体肌腱材料。

[Objective] To explore the effect of different entrainers on tendon allografts treated by supercritical CO2 fluid extraction(SCCO2). [Methods] The tendon allografts were randomly divided into four groups, The tendons in the Group A were treated by freezing only without SCCO2, while those were treated by SCCO2 with 100% ethanol as entrainer in Group B, SCCO2 with ethanol+1% ethylene diamine tetraacetic acid(EDTA) in Group C and SCCO2 with ethylene oxide/propylene oxide polyether glycol(EO/PO) as entrainer in the Group D.The tendons in the four groups were assessed by mechanical testing, histological examination, and toxicity test with cell proliferation in vitro. [Results] As consequence of mechanic tests, there was no significant difference in the maximum load among the four groups(P>0.05).However, the anti-pulling strength decreased(P>0.05), whereas elastic modulus significantly decreased and the elongation at break significantly increased in Group D compared with those in Group A(P<0.05), while which remained unchanged in Group B and C in contrast with the group A(P>0.05). The histological examination revealed that the extracellular matrix remained unchanged while cellular component vanished completely in the Group D, by contrast, a great deal of cellular nucleus seen in Group A. In term of cellular proliferation in vitro for toxicity test, there were no statistical differences in OD value of cell proliferation among the 4 groups(P>0.05). [Conclusions] The ideal decellularized tendon allograft with no cellular toxicity might be obtained by SCCO2 treatment with(EO/PO) as the entrainer.