试剂对脱钙骨基质成骨能力影响的实验研究

The influence of defatting and decalcifying solutions on the matrix-induced bone formation

目的 :研究异体骨基质制备过程中常用的脱脂和脱钙试剂溶液对其成骨能力的影响。方法 :8周鼠龄的Wistar雌鼠作为供体 ,切取胫股骨 ,碾磨成 12 5～ 80 0微米的颗粒 ,分成四组 ,分别用丙酮 -盐酸、丙酮 -乙二胺四乙酸 (EDTA)、三氯甲烷 -甲醇 -EDTA或三氯甲烷 -甲醇 -盐酸 (控制组 )脱脂和脱钙。冷冻干燥处理后 ,每 3 0mg装入一胶囊中。 60只 6周鼠龄的Wistar雌鼠作为受体 ,植入腰背肌中 ,每只鼠接受同一组植入材料 2枚。术后 6周取标本测定重量、干燥后的重量、钙含量、碱性磷酸酶含量和组织形态学定量测定。结果 :丙酮 -盐酸组的干燥重量 (P <0 .0 1)和钙含量 (P <0 .0 5 )明显高于控制组。结论 :用丙酮清除脂质的异体骨基质成骨能力优于用三氯甲烷 -甲醇混合液。EDTA或盐酸脱钙对其成骨能力无明显影响。

Objective:To investigate the effect of some defatting and decalcifying solutions on the capacity of the matrix induced bone formations. Methods:The pulverized bones, which were 125～800 micron, from eight week old Wistar female rats were divided into four groups. The particles defatted and decalcified separately with acetone HCl, acetone EDTA, chloroform methanol EDTA served as experimental groups while chloroform methanol HCl as control group. The demineralized bone matrixes (DBM) were lyophilized. 60 Wistar female rats acted as receptors. Each 30mg DBM was implanted in the paravertebral muscles. Evaluations were assessed by the explanted weight ratio, calcium and alkaline phosphatase contents and histomorphometric measures 6 weeks postoperatively. Results:The acetone HCl group was significantly greater than the control when the final dried weight ( P <0.01) and calcium content ( P <0.05) were measured. Conclusion:The lipids extraction of DBM with acetone solution has more osteoinductive capacity than that with chloroform methanol solution. No different influence on matrix induced osteogenesis occurrs when the bone demineralized with either EDTA or HCl.