组织工程化骨修复四肢骨缺损的初步临床研究

A Preliminary Experimental Study on Repairing Defects of Extremities Using Tissue Engineer Bone

目的探索以人自体骨髓基质干细胞(Human bone marrow stromal cells,hBMSCs)为种子细胞构建的组织工程骨用于修复四肢骨缺损的可行性。方法选择9例四肢骨缺损病例(良性骨肿瘤5例、骨缺损4例)。经病人髂前上嵴,穿刺抽取10mL骨髓,用密度梯度离心法分离得到hBMSCs,用成骨诱导培养液定向培养、扩增,应用第3代的hBMSCs与珊瑚复合,继续在体外培养1周后,手术植入骨缺损区。分别在术后2个月、6个月、12个月和24个月进行临床观察和X线检查。1例病人在二次手术时,在原骨缺损区取出少量新生组织活检,进行组织学观察(HE染色)。结果患者术后X线检查显示,组织工程骨在原位形成新骨,能较好地修复骨缺损。活检标本HE染色显示,局部形成正常松质骨组织。结论以自体hBMSCs为种子细胞,利用组织工程技术制成的自体组织工程骨可以修复四肢骨缺损,并且能够修复较大的骨缺损,但组织工程骨的临床成骨效果与植骨床血供密切相关,目前所使用的组织工程骨植骨方法具有一定的适应证。

Objective To explore the feasibility of human bone marrow stromal cells （hBMSCs） and its clinical repair of limb bone defects. Methods Nine patients of bone defects （5 bone tumors, 4 bone defects） were included in this study. The hBMSCs were isolated by Percoll gradient centrifugation with bone marrow aspirated from iliac crest of each patient, hBMSCs were cultured in vitro and induced to become osteogenic cells in the DEME medium containing 10% self-serum, β- glycerophosphate （10 nmol/L） dexamethasone （10-8 mol/L）, L-2-aseorbic acid （50 μmol/L）, and 1-25（OH）2VD3 （10 nmol/L）. Induced hBMSCs of third passage were harvested and seeded into coral to form cell-scaffold construct and in vitro co-culture for one week. The bone defects were repaired with the cell-scaffold construct. All cases were followed up by X-ray and clinical examination at 2, 6, 12, 24 months post-operation. In one case who received second surgery, a piece of tissue was harvested at the implantation site for histological examination. Results The X-ray examination showed that bone defects were repaired well. HE staining revealed the structure of new-born tissue were similar to that of normal bone. Conclusion Engineered bone can be used to repair clinical bone defect of limbs, and can repair large bone defects. But the bone-forming of tissue engineered bone is relative to the blood supply in bone implanting bed.