摘要
目的:运用自行建立的血管脱细胞方法,制备人股动脉同种异体血管移植材料。方法:实验于2004-09/2005-09在空军总医院临床实验中心完成。①取材:股动脉供者为急性外伤后右下肢截肢患者,男,26岁,患者及家属同意捐献截除的右下肢,并经过医院伦理道德委员会许可。右下肢离体至取材时间为6h,取材在无菌条件下进行。②人股动脉血管脱细胞化学萃取:①无离子水(低渗)12h→②10g/LTriton24h→③磷酸盐缓冲液洗涤3次后,无离子水4h→④10g/LSDS24h→⑤用磷酸盐缓冲液洗涤3次后,无离子水4h,用含抗生素的Hanks溶液漂洗2次,4h/次。上述各步中均加入蛋白酶抑制剂苯甲基磺酰氟5μmol/L。③力学特性测量及组织学观察:选择其中两根脱细胞血管测量二维爆破压。并通过苏木精-伊红、V-W等染色方法观测细胞脱出情况和脱细胞后胶原和弹性纤维情况,并通过扫描电镜观察脱细胞股动脉的内膜超微结构。结果:①经去垢剂10g/LTritonX-100,10g/LSDS等多步骤脱细胞方法处理后,人股动脉血管的细胞基本脱除,剩余成分主要为维持血管张力的胶原蛋白和弹性蛋白,细胞外基质和血管的弹性、韧性保持完好,可在正常张力下行常规的血管缝线吻合。②两根脱细胞血管的二维爆破压分别为950mmHg(1mmHg=0.133kPa)和>1000mmHg;远高于人体血压,可以满足移植的需要。③脱细胞血管的内膜表面内皮细胞已不复存在,断面上可见纤维组织排列疏松不够整齐,组织之间可见细小间隙。结论:经去垢剂TritonX-100,SDS加低渗溶液和蛋白酶抑制剂处理的多步脱细胞法可以脱除人股动脉血管的细胞成分,细胞外基质和血管的弹性、韧性保持完好,制备出的人脱细胞股动脉可以作为同种异体血管移植材料。
AIM： To set up a new process to access the preparation of human acellular femoral artery allograft.
METHODS： The experiment was accomplished in the Clinical Experimental Center of Air Force General Hospital of Chinese PLA between September 2004 and September 2005. （1）Material source： The femoral artery of right limbs were donated by one male amputation patient after acute trauma, aged 26 years and agreed to donation, with the approval of ethic and morality committee. The excision was carried out sterily within 6 hours after isolation.（2）Chemical extraction： A multi-step process, including hypotension of deionized water for 12 hours, combination with 10 g/L 1% t-octyl-phenoxypolyethoxyethanol （Triton X-100）, and sodium dodecyl sulfate （SDS） detergents for 3 times, then deionized water for 4 hours, 10 g/L SDS for 24 hours, and phosphate buffer for 3 times, which were followed by deionized water for 4 hours and washed by Hanks solution containing antibody for twice, 4 hours each were performed in human acellular femoral artery vascular added with 5 μmol/L phenylmethylsulfonyl fluoride. （3）Mechanics feature measurement and histological examination： Two-dimensional blasting pressure was assessed in two decellularized vessels. Hematoxylin-eosin and V-W staining were applied to determine the collagen and elastic fiber after decellularization. Scan electron microtelescope was done to observe the intimal ultrastructure on the samples.
RESULTS： （1）A multi-step process with 10 g/L SDS and 10 g/L Triton X-100 could remove all cells with the extracellular matrix well conserved in human acellular femoral artery, remaining the collagen and elastin to maintain angiotasis and present elasticity and toughness well. And vessel could be sutured well under normal tension. （2） Two-dimensional blasting pressure of two decellularized vessels were 950 mm Hg （1 mm Hg=0.133 kPa） and 〉 1 000 mm Hg, respectively, which were higher than human blood pressure and satisfied the transplantation.�
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第16期3022-3025,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
