制备去细胞人体肌腱支架材料的实验研究  

Experimental study of preparing aceilular human tendon scaffold by chemical extraction methods

作　　者：许银峰[1] 胡军[1] 肖良宝[1] 江丽[1] 易建华[1] 牛晓锋[1] 

机构地区：[1]中山大学附属第一医院黄埔院区,广州510700

出　　处：《中华显微外科杂志》2013年第2期144-148,共5页Chinese Journal of Microsurgery

基　　金：广东省科技计划项目（20108031100005）;广东省自然科学基金（10451008901005905）;国家科学技术部“863”科技攻关项目（2006AA02A130）;广州市科技攻关重大项目（200521-E0091）

摘　　要：目的探索化学萃取法制备去细胞人体肌腱支架材料的实验方法。方法用1．0％磷酸三丁酯[tri（n—butyl）Phosphate，TnBP]分别联合0％、0．25％、0．50％及1．00％曲拉通-100（TritonX-100）对人体肌腱进行去细胞处理，通过组织学、扫描电镜观察、生物力学和羟脯氨酸含量测定，并与新鲜人体肌腱对比，评价去细胞人体肌腱支架材料的特性。结果肌腱去细胞处理后色泽光亮，腱膜完整，韧性好；1．00％TnBP加0％TritonX-100组仍有少量细胞存留，其余3个处理组去细胞彻底，1．00％TnBP加0．25％TritonX-100组胶原纤维完整、走形规则。1．00％TnBP加0．50％TritonX-100与1．00％TnBP加1．00％TritonX-100组胶原纤维完整，但间隙略增宽，最大载荷分别为（385．220±80．316）N、（398．220±127．195）N，抗张强度分别为（46．690±16．295）Mpa、（46．20±5．517）Mpa，羟脯氨酸含量分别为（0．2826630±0．0110109）μg/mg、（0．2793550±0．0102129）μg/mg，分别和新鲜对照组[最大载荷（533．280±135．774）N、抗张强度（65．560±14．401）Mpa、羟脯氨酸含量（0．2928820±0．0100988）μg/mg]相比，力学性能降低、羟脯氨酸含量减少，差异有统计学意义（P〈0．05）。结论1．oo％TnBP加0．25％Tritonx-100处理是较理想的制备去细胞人体肌腱的方法。Objective To explore a method of preparing the acellular tendons of human with chemical approaches. Methods From April 2011 to June 2012, several treatments were performed on human tendons using 1.00% tri（n-butyl） Phosphate （TnBP） combinded with Triton X-100 at the concentration of 0, 0. 25%, 0. 50% , 1.00% respectively. Specimens were examined using histological observation, scanning electron mi- croscopy, biomechanical testing and hydroxyproline quantitation. The results were then compared with fresh ten-dons of control group, so as to value the characteristics of decellularized human tendon scaffold. Results A- cellular human tendons had glossy surface, intact aponeurotic membrane and satisfactory flexibility. A small number of disrupted ceils remained in the 1.00% TnBP ＋ 0% Triton X-100 treated tissue, while other three ex-perimental groups successfully eliminate all cells. Intact and regular collagen architecture was retained in 1.00% TnBP ＋0. 25% Triton X-100 treated tissue. 1.00% TnBP ＋0. 50% Triton X-100 and 1.00% TnBP ＋ 1.00% Triton X-100 treated tissue were nearly identical to 1.00% TnBP ＋0. 25% Triton X-100 treated tissue, but the interval of collagen was slightly wider than the control group, the maximum load （ 385.22 ± 80. 32N, 398.22 ± 127.20N） , ultimate tensile strength（46. 69 ± 16. 30Mpa, 46. 20 ± 5.52Mpa）and hydroxyproline con-tent（0. 282663 ±0. 0110109 μg/mg, 0. 279355 ±0. 0102129 μg/mg） were statistically lower （P 〈0. 05） com-pared with those of the eontrol group, maximum load （ 533.28 ± 135.77 N ） , ultimate tensile strength （ 65.56 ± 14. 40Mpa） and hydroxyproline content （ 0. 292882 ± 0. 0100988 μg/mg） respectively. Conclusion The de-cellularization treatment with 1.00% TnBP ＋ 0. 25% Triton X-100 could be optimized for preparing acellular hu-man tendons.

关 键 词：肌腱 化学萃取 去细胞 磷酸三丁酯 曲拉通-100 

分 类 号：R6[医药卫生—外科学] R3[医药卫生—临床医学]