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The Development for Osteoinductive and Biodegradable Bone Substitute Materials

骨诱导和可生物降解骨替代材料的开发

基本信息

批准号：
03557086
负责人：
ENOMOTO Shoji
金额：
$ 7.81万
依托单位：
The Department of Oral and Maxillofacial Surgery 2, Faculty of Dentistory, Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

项目摘要

There are many cases to make bone defects in the oral and maxillo-facial region, for example cysts, cancers and so on. So Bone Morphogenetic Protein (BMP) is needed for clinical usage. And a lot of studies have been done to purify the BMP and to investigate the physiological function of that. Several reports suggested the possibility that BMP (Osteogenin, BMP-3 from demineralized bone matrix have stronger osteoinductive activity rather than rhBMP -2. Therefore the BMP was purified from the demineralized bone extract by gel filtration on Sephacry1 S-200HR, cation exchange with Mono-s, Heparin affinity column and finally by c1/8 reverse phase chromatography. The BMP preparation obtained in this study induced ectopic bone and marrow tissue in ectopic site in 2 weeks after implantation into the thoracic subcutaneous tissue. The results of 15% SDS-PAGE revealed the molecular weight in reduced condition is half that in non-reduced condition as reported before. Peptide sequence analysis revealed that the purified BMP fraction contained "BMP-3" reported by Wozney et al.A skull defect which is 8 mm in diamenter was prparated, and porus poly-L-lactic acid disc including the BMP was implanted into the defect. Osteoinductive activity induced by the disc was evaluated by means of soft X-ray image, calcium content, and histological observation. 2 weeks after implantation, bone tissue was induced and large value of calcium was detected in the excised discs. 4 weeks after implantation, also bone tissue was induced and larger value of calcium was detected from the excised discs. These results supported that the porus poly-L-lactic acid disc will be effective for recovery of bone defect.

口腔颌面部骨缺损的情况很多，如囊肿、肿瘤等，临床上需要骨形态发生蛋白（BMP）。人们对BMP的纯化及其生理功能进行了大量的研究。一些报道认为BMP（Osteogenin，脱钙骨基质中的BMP-3）可能比rhBMP-2具有更强的骨诱导活性。因此，通过Sephacry 1 S-200 HR凝胶过滤、Mono-s阳离子交换、肝素亲和柱以及最后通过c1/8反相色谱法从脱矿骨提取物中纯化BMP。本研究中获得的BMP制剂植入胸部皮下组织后2周诱导异位部位的异位骨和骨髓组织。15% SDS-PAGE结果显示，还原条件下的分子量为非还原条件下的一半。肽段序列分析表明，纯化的BMP组分中含有Wozney等报道的“BMP-3”。制备直径为8 mm的颅骨缺损，将含有BMP的多孔聚L-乳酸圆盘植入缺损处。通过软X线图像、钙含量和组织学观察评价椎间盘诱导的骨诱导活性。植入后2周，诱导骨组织，并在切除的椎间盘中检测到大量的钙。植入后4周，也诱导骨组织，并从切除的椎间盘中检测到较大的钙值。这些结果支持多孔聚-L-乳酸盘将有效地用于骨缺损的修复。