脱细胞血管基质制备和异体移植的实验研究

Experimental study on preparation of decellularized artery vascular graft matrix and explantation of carotid artery allografts

目的研究建立无细胞血管基质制备的新方法,并探讨脱细胞血管异体移植的可行性。方法采用不同去垢剂〔1%TritonX100、1%十二烷基磺酸钠(SDS)〕多步骤对血管进行脱细胞处理,并通过组织学和力学观测,建立兔动脉血管脱细胞的方法;并进行脱细胞血管的异体移植。结果经低渗溶液和去垢剂1%TritonX100、1%SDS等多步骤处理,兔胸主动脉血管的细胞基本脱除,细胞外基质保持完好,血管的弹性、韧性和力学强度仅轻度降低;用该法制备的兔颈动脉(直径约2.0mm)进行异体移植,经2个月观察,全部通畅。结论经去垢剂TritonX100、SDS加低渗溶液和蛋白酶抑制剂处理的多步法,可以脱除血管的细胞成分,细胞外基质和力学特性保持完好,是一种较好的方法;用该法制备的兔颈动脉(直径约2.0mm)进行异体移植,初步取得成功。

Objective To set up a new process to access the preparation of decellularized artery grafts. And to evaluate the feasibility of decellularized artery allografts was evaluated. Methods This study compared the effects of four extraction chemicals [1% t-octyl- phenoxypolyethoxyethanol (Triton X-100), 1%tri(n-butyl)phosphate (TnBP),and 1% sodium dodecyl sulfate (SDS) and trypsin(0.125, 0.25%) on thoracic artery vascular for 24 h (except trypsin for 2 h). At the base of it, a four-step process, including hypotonic, hypertonic solutions and combining with 1% Triton X-100 and 1% SDS detergrnts, were performed in rabbit thoracic artery vascular. Histological examination, tensile tests and expanding-burst tests were done on the samples. The decellularized carotid artery allografts were transplanted in other rabbits. Results Treatment with 1% SDS or 1% Triton X-100 for 24 h could remove most cells with retention of near normal structure. A four-step process could remove all cells with the extracellular matrix well conserved. The pulling mechanical properties and burst pressure of decellularized carotid artery were similar to the control. The decellularized carotid artery allografts(diameter of 2 mm) were patent at explanting up to 2 months. Conclusions The acellular artery vascular graft matrix is well prepared with four-step process including detergents, such as TritonX-100、SDS without compromising the graft structure or mechanical properties significently. The carotid artery allografts(diameter of 2 mm) decellularized by the process are patent at explanting up to 2 months. [