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目的 观察贮存时间对天然牛骨形态发生蛋白 (bBMP)诱导成骨活性及抗原性的影响。方法 取 1989～ 1998年提取并于 4℃贮存的 6个批次的 bBMP，以 SDS-PAGE凝胶电泳法测定其相对分子质量。 108只 Balb/c雄性小鼠被分为 6组，每组 18只，按 Urist方法于每只鼠股部肌间隙肌袋内植入 2 mg bBMP，术后第 7、第 14及第 28 d取材，以间接免疫荧光染色法分别测定抗体滴度及阳性染色率；以酶动力学方法测定碱性磷酸酶（ ALP）活性，并观察 bBMP诱导成骨活性。结果 (1)SDS-PAGE凝胶电泳显示， 8条电泳带均为多肽片段， 6个批次 bBMP的相对分子质量集中在 14× 10 3、 18× 10 3、 25× 10 3、 30× 10 3和 40× 10 3，各批次区带分布基本一致。 (2)间接免疫荧光染色测定显示， bBMP可刺激小鼠产生弱的荧光抗体，抗体滴度 1∶ 2～ 1∶ 4，随着贮存时间的增加荧光抗体的阳性染色率有所下降，但差异无显著性意义 (P >0.05)。 (3)植入 bBMP后第 7、第 14及第 28 d各批次 ALP活性明显高于对照组 (均 P0.05)。 (4)组织学观察，在 bBMP植入第 7 d即可见大量间充质细胞分化，并向软骨细胞转化；第 14 d，有大量骨软骨形成；第 28 d，见成熟板层骨，并有骨髓形成。结论 于 4℃贮存 10年的 bBMP仍具有良好的诱导成骨活性，植?
Objective To observe the effect of storage time on bBMP induced osteogenic activity and antigenicity. Methods Six batches of bBMP extracted from 1989 to 1998 and stored at 4 ℃ were determined by SDS-PAGE gel electrophoresis. A total of 108 Balb / c male mice were divided into 6 groups with 18 rats in each group. 2 mg bBMP was implanted into the intramural muscle space of each rat according to the Urist method. At the 7th, 14th and 28th day after operation, The antibody titers and positive staining rates were determined by indirect immunofluorescence staining. The alkaline phosphatase (ALP) activity was measured by enzyme kinetics and the osteoblast activity induced by bBMP was observed. Results (1) SDS-PAGE gel electrophoresis showed that the eight electrophoresis bands were all polypeptide fragments. The relative molecular mass of the 6 batches of bBMP was 14 × 10 3, 18 × 10 3, 25 × 10 3 and 30 × 10 3 and 40 × 10 3, the distribution of each batch basically the same. (2) Indirect immunofluorescence staining showed that bBMP could stimulate mice to produce weak fluorescent antibody with antibody titers of 1: 2 to 1: 4. With the increase of storage time, the positive staining rate of fluorescent antibody decreased, but the difference No significant (P> 0.05). (3) ALP activity in batches of 7th, 14th and 28th day after bBMP implantation were significantly higher than those in control group (all P <0.05). (4) Histological observation showed that a large number of mesenchymal cells were differentiated and transformed into chondrocytes on the 7th day after bBMP implantation. On the 14th day, there was a large amount of osteochondral formation. On the 28th day, mature lamellar bone was seen Bone marrow formation. Conclusion bBMP stored at 4 ℃ for 10 years still has good osteogenic activity,