新鲜骨髓吸出物直接种植于支架构建组织工程韧带

Seeding fresh bone marrow aspirate directly on scaffolds to construct tissue-engineered ligament

背景:有关于应用新鲜骨髓吸出物直接局部注射修复韧带部分损伤的报道,少有提及应用新鲜骨髓吸出物直接种植于支架构建组织工程韧带的研究报道。目的:评价将新鲜骨髓吸出物直接种植于支架构建组织工程韧带的可行性。方法:构建丝素纤维/小肠黏膜下层复合支架后,骨髓种植组将骨髓吸出物直接种植于复合支架上;细胞种植组将骨髓间充质干细胞种植于复合支架上;以未接种任何物质的复合支架作为空白对照,检测各组细胞黏附密度、细胞增殖活性及细胞外基质分泌情况。将骨髓种植组复合物植入兔前交叉韧带横断处,12周后取材评价骨髓种植组材料体内生物相容性。结果与结论:骨髓种植组孵育4 h后的细胞黏附密度、不同时间点细胞增殖率显著高于细胞种植组(P<0.05)。骨髓种植组及细胞种植组Ⅰ型、Ⅲ型胶原分泌量均显著高于空白对照组(P<0.05),但骨髓种植组及细胞种植组两组间Ⅰ型、Ⅲ型胶原分泌量差异无显著性意义。骨髓种植组材料植入兔体内未引起致死性免疫排斥反应及严重炎症反应,未见明显韧带再生及血管化。说明新鲜骨髓吸出物直接种植于支架可构建组织工程韧带,并且体内短期生物相容性良好。

BACKGROUND：There are several reports about the application of fresh bone marrow aspirate being injected directly to repair partial ligament injury, but the application about fresh bone marrow aspirate directly being planted on scaffolds to build tissue-engineered ligament is rarely mentioned. OBJECTIVE：To evaluate the feasibility of applying fresh bone marrow aspirate planted directly on scaffolds to construct tissue-engineered ligament METHODS：We constructed fibroin fiber/smal intestinal submucosa composite scaffold, then planting fresh bone marrow directly to built bone marrow seeding group and planting seed cel s （bone marrow mesenchymal stem cel s） on the scaffold to built cel seeding group. The control group had no treatment. After that, we detected the density of cel adhesion, cel proliferation ability and extracel ular matrix secretion. Then, the composite in the bone marrow seeding group was implanted into the broken anterior cruciate ligament in rabbits, and material biocompatibility in vivo was evaluated after 12 weeks. RESULTS AND CONCLUSION：After 4 hours of incubation, bone marrow seeding group was significantly higher than the cel seeding group in cel adhesion density and proliferation rate （P＜0.05）. Bone marrow seeding group and cel seeding group showed higher type I, III col agen secretion compared with the control group （P＜0.05）, but the col agen secretion of bone marrow seeding group and cel seeding group showed no significant difference. Composite cel scaffold implantation in vivo did not cause fatal immune rejection and severe inflammatory＆amp;nbsp;reaction, and no significant ligament regeneration and vascularization occurred. These findings indicate that fresh bone marrow aspirate can be seeded directly on scaffolds to construct tissue-engineered ligament, and the short-term biocompatibility in vivo is good.