Ⅰ型胶原海绵／血管内皮生长因子缓释材料的组织相容性研究

Biomechanics of type Ⅰ collagen/vascular endothelial growth factor slow-release material

目的检测Ⅰ型胶原（CI）海绵／血管内皮生长因子（VEGF）缓释材料的组织相容性。方法将兔骨髓间充质干细胞（BMSCs）分别接种到Ⅰ型胶原（CI）、Ⅰ型胶原．血管生长因子（CI＋VEGF）海绵材料及普通细胞培养板上，于第1～10天每天进行细胞计数试剂盒（CCK-8）法检测，于第5天进行扫描电镜观察。将1cm^2 CI海绵及CI＋VEGF缓释海绵材料分别植入兔皮下，于第7、14、21、28天取材进行苏木素-伊红（HE）染色法分析。结果细胞计数试剂盒（CCK-8）分析显示在各个检测时间点，BMSCs增殖程度CI＋VEGF组〉CI组〉单纯培养组，第4天3组细胞活力分别为2．77±0．21、2．21±0．17、1．86±0．13（P〈0．05）；第8天分别为2．47±0．19、1．89±0．15、1．16±0．11（P〈0．05）。扫描电镜观察：有大量BMSCs黏附生长在CI＋VEGF海绵支架表面及孔隙壁。HE结果显示CI＋VEGF缓释海绵材料与CI具有相似的炎性反应，28d基本完全降解。结论CI＋VEGF缓释海绵材料具有良好组织相容性。

Objective To investigate the histocompatibility of the type Ⅰ collagen （ CI）/vascular endothelial growth factor （VEGF） slow-release material. Methods Bone marrow-derived mesenchymal stem cells （BMSCs） were obtained with passage isolation, and then seeded on cross-linked CI, CI ＋ VEGF materials and cell cuhure plate. Cytoeompatibility was detected by using scanning electron microscopy （SEM） at day 5 and cell counting kit-8 （CCK-8） test at every day during 10 days after seeding. The CI and CI ＋ VEGF materials were subcutaneously implanted into 8 New Zealand albino rabbits. The rabbits were sacrificed at 7, 14, 21 and 28 days and the scaffolds and the surrounding tissues were extracted for hematoxylin-eosin （HE） staining. Results CCK-8 test revealed that the cell proliferation in 3 groups was as follows ： CI ＋ VEGF group 〉 CI group 〉 common culture group, and the cell viability in CI ＋ VEGF group was significantly higher than that in controls （ day 4 ： 2. 77 ± 0. 21 〉 2. 21 ± 0. 17 〉 1.86 ± 0. 13 P 〈 0. 05 ; day 8：2. 47 ± 0. 19 〉 1.89 ± 0. 15 〉 1.16 ± 0. 11, P 〈 0. 05 ）. SEM image of the scaffolds showed that BMSCs spread well and homogenously distributed throughout the entire scaffolds. Histological FIE staining showed that the immune rejection of CI ＋ VEGF sponge material was similar to CI, and CI ＋ VEGF sponge material could be degradated at 28 days in animals. Conclusion CI ＋ VEGF sponge materials have well histocomnatibilitv.