小肠黏膜下层-蚕丝复合前交叉韧带重建支架的初步研究  

A PRELIMINARY STUDY ON SMALL INTESTINAL SUBMUCOSA-SILK COMPOSITE SCAFFOLD TO RECONSTRUCT ANTERIOR CRUCIATE LIGAMENT

作　　者：曹杰[1,2] 王旻[3] 解慧琪[3] 魏人前[1,2] 

机构地区：[1]南方医科大学第三临床医学院,广州510515 [2]佛山市第二人民医院骨科 [3]四川大学华西医院生物治疗国家重点实验室.干细胞与组织工程研究室

出　　处：《中国修复重建外科杂志》2015年第12期1534-1540,共7页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：广东省医学科研基金资助项目(A2010601)~~

摘　　要：目的制备小肠黏膜下层(small intestin al submucosa,SIS)-蚕丝复合支架用于前交叉韧带(anterior cruciate ligament,ACL)重建,并对其力学性能、细胞相容性、组织相容性以及体内重建减缓胫骨隧道关节液渗入作用进行评价。方法蚕丝脱胶后编织成蚕丝支架,将膜状SIS呈圆筒状包绕在蚕丝支架外围构建复合支架,采用生物力学测试仪评价支架生物力学性能;支架复合BMSCs培养后,以活死细胞染色及细胞计数试剂盒8法评价支架的细胞相容性。取30只6周龄SD大鼠随机分为蚕丝支架组(S组)与复合支架组(SS组),每组15只,分别将两组支架材料移植于大鼠皮下,2、4、8周取材行HE染色观察组织相容性。取20只28周龄新西兰大白兔双侧后肢随机分为S组与SS组(n=20),分别用蚕丝支架与复合支架行双膝ACL重建,胫骨隧道开窗,5 m L 10%Na Cl和墨水溶液分别行关节腔内注射,不同时间点测量骨窗内移植物的电阻值并记录骨窗处墨水渗出时间。结果支架大体观察见复合支架内部为双股螺旋蚕丝束,外围SIS呈圆筒状包绕。蚕丝支架与复合支架最大载荷分别为(138.62±11.41)、(137.05±16.95)N,刚度分别为(24.65±2.62)、(24.21±2.39)N/mm,差异均无统计学意义(P>0.05)。活死细胞染色显示蚕丝支架和复合支架上细胞活力均良好,其中复合支架上细胞伸展性更好。细胞增殖曲线示蚕丝支架组、复合支架组及正常细胞对照组各时间点吸光度(A)值差异均无统计学意义(P>0.05)。大鼠皮下埋植标本HE染色观察示,材料植入后SS组与周围组织相容性好,炎性细胞和新生血管数与S组比较,差异均有统计学意义(P<0.05)。兔关节腔灌注后SS组电阻值开始下降时间点和墨水渗出时间均明显晚于S组(P<0.05),且墨水渗出持续时间明显长于S组(P<0.05)。结论制备的SIS-蚕丝复合支架生物力学性能及细胞、组织相容性好,体内移植能早期血管化,并对ACL重建早期Objective To prepare the small intestinal submucosa （SIS）-silk composite scaffold for anterior cruciate ligament （ACL） reconstruction, and to evaluate its properties of biomechanics, biocompatibility, and the influence on synovial fluid leaking into tibia tunnel so as to provide a better choice in the clinical application of ACL reconstruction. Methods The silk was used to remove sericin and then weaved as silk scaffold, which was surrounded cylindrically by SIS to prepare a composite scaffold. The property of biomechanics was evaluated by biomechanical testing system. The cell biocompatibility of scaffolds was evaluated by live/dead staining and the cell counting kit 8 （CCK- 8）. Thirty 6-week-old Sprague Dawley rats were randomly assigned to 2 groups （n=15）. The silk scaffold （S group） and composite scaffold （SS group） were subcutaneously implanted. At 2, 4, and 8 weeks after implanted, the specimen were harvested for HE staining to observe the biocompatibility. Another 20 28-week-old New Zealand white rabbits were randomly assigned to the S group and SS group （n=20）, and the silk scaffold and composite scaffold were used for ACL reconstruction respectively in 2 groups. Furthermore, a bone window was made on the tibia tunnel. At last, the electric resistance of tendon graft in the bone window was measured and recorded at different time points after 5 mL of 10% NaC1 or 5 mL of ink solution was irrigated into the joint cavity recspectively. Results The gross observation showed that the composite scaffold consisted of the helical silk bundle inside which was surrounded by SIS. The maximal load of silk scaffold and composite scaffold was respectively （138.62_＋11.41） N and （137.05~16.95） N, showing no significant difference （P〉0.05）; the stiffness was respectively （24.65＋2.62） N/mm and （24.21_＋2.39） N/mm, showing no significant difference （P〉0.05）. The live/dead staining showed that the cells had good activity on both scaffolds. However, the cells on the c

关 键 词：前交叉韧带重建 小肠黏膜下层 蚕丝 复合支架 大鼠 兔 

分 类 号：R318.08[医药卫生—生物医学工程]