Mass spectrometric identification of intrinsic growth factors accumulated with Immaterial and reconstruction of highly activated bone regenerative material

将磷酸八钙（OCP）植入标准化大鼠颅骨缺损。植入10天后，取出植入物进行冻干，提取随植入物积累的蛋白质。对提取的蛋白进行纯化，并采用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）测定。虽然从实验组中检测到200多种蛋白质，但很难确定哪种蛋白质直接参与骨再生。在研究过程中，我们制备了骨形态发生蛋白-2 （BMP-2）复合OCP (OCP/BMP-2)，并将其植入大鼠颅骨缺损。由于OCP/BMP-2可协同促进骨再生，OCP可作为BMP-2的有效支架[J] .中国生物医学工程杂志，2001,19(1):29 - 31。然后，我们设计了OCP-胶原复合材料（OCP/Col）作为骨组织的仿生成分，并进行了植入。OCP/Col比单独使用OCP显著增强骨再生（J Biomed Mater Res B apple Biomater 79B: 210-217, 2006），申请了“替代硬组织支架”专利。此外，我们证实在大鼠颅骨缺损中植入OCP/Col比β-磷酸三钙胶原复合材料或羟基磷灰石胶原复合材料更能促进骨再生，这表明OCP/Col将是一种有效的骨再生材料（J Biomed Mater Res a in press, 2007）。

Octacalcium phosphate (OCP) was implanted into the standardized rat calvarial defect. After ten days of implantation, the implants were retrieved and lyophilized, then the protein accumulated with the implants were extracted. The extracted protein was purified and was determined by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Although over 200 types of protein were detected from the experimental groups, it was difficult to identify which protein was directly involved in bone regeneration.During the process of this investigation, we prepared a composite bone morphogenetic protein-2 (BMP-2) combined with OCP (OCP/BMP-2) and implanted into the rat calvarial defect. Because OCP/BMP-2 synergistically enhanced bone regeneration, OCP may be useful as an effective scaffold for BMP-2 (J Biomed Mater Res A 71A : 299-307, 2004). Then, we have engineered OCP-collagen composite (OCP/Col) that was biomimetic components of bone tissue, and implanted. OCP/Col significantly enhanced bone regeneration than OCP alone (J Biomed Mater Res B Appl Biomater 79B : 210-217, 2006), and a patent as "Substitutional scaffold for hard tissue" was applied.. Furthermore, we confirmed that the implantation of OCP/Col in a rat cranial defect enhanced more bone regeneration than β-tricalcium phosphate collagen composite or hydroxyapatite collagen composite, suggesting that OCP/Col would be an effective bone regenerative material (J Biomed Mater Res A in press, 2007).