人纳米脱钙骨基质复合物的理化性质和安全性

背景：脱钙骨基质和骨形态发生蛋白已被证实具有良好的骨诱导性,但有关纳米脱钙骨基质的研究较少,其理化性质和生物安全性尚不明确。 目的：在前期实验制备人纳米脱钙骨基质的基础上加载重组人骨形态发生蛋白2,分析人纳米脱钙骨基质复合重组人骨形态发生蛋白2的理化性质及生物安全性。 方法：采用改良Urist法制备人脱钙骨基质,并进行纳米化处理,再将骨形态发生蛋白2与其按特定比例混合,冻干塑型行以下实验：①热源实验：将材料浸提液经耳静脉注入兔体内。②毒性实验：将材料浸提液与生理盐水分别经尾静脉注入小白鼠体内。③植入实验：在兔两侧后肢肌肉内分别植入实验材料和β-磷酸三钙。 结果与结论：冻干塑型后,纳米人脱钙骨基质材料表面致密,孔隙直径100-400 μm,孔隙分布欠均匀,孔隙率小于30%,以碳、氧和氮为主要元素组成。人纳米脱钙骨基质复合重组人骨形态发生蛋白2材料无热源效应,注射后未见兔体温有明显波动。急性全身毒性实验结果表明人纳米脱钙骨基质复合重组人骨形态发生蛋白2材料符合国家相关规定,注射后未见小鼠出现明显毒性反应。人纳米脱钙骨基质复合重组人骨形态发生蛋白2材料植入兔体内的炎症反应明显轻于β-磷酸三钙植入后的反应。结果表明人纳米脱钙骨基质复合重组人骨形态发生蛋白2是一种无毒、组织相容性好、生物利用度高、炎症反应轻的纳米同种异体骨移植替代物。

Physicochemical property and safety of nanometer human demineralized bone matrix composite

BACKGROUND: Demineralized bone matrix and bone morphogenetic protein have been shown to have good bone induction, but less studies concerned nanometer demineralized bone matrix. Its physical and chemical properties and biological security are not yet clear. OBJECTIVE: On the basis of preparing the nanometer human demineralized bone matrix in previous experiment, we mixed the recombinant human bone morphogenetic protein-2 together to obtain the new bone graft substitute and to research its physical and chemical properties and biological security. METHODS:The human demineralized bone matrixes were prepared by the method of modified Urist and nano-processed then mixed with the bone morphogenetic protein-2 in specific proportions in order to be lyophilized to complete the following experiments. (1) Pyrogen experiment: the material extracts were injected in the rabbits by ear intravenous. (2) Toxicity experiments: material extracts and saline were separately injected via the tail vein of mice in vivo. (3) Implantation experiments: experimental materials and β-tricalcium phosphate were implanted into rabbits on both sides of the hindlimb muscle. RESULTS AND CONCLUSION: After lyophilized shaping, the nanometer demineralized bone matrix material had dense surface and it’s pore diameter was 100-400 μm. The pore distribution was less uniform and the porosity was of less than 30%. The main elements were carbon, oxygen and nitrogen. Nanometer human demineralized bone matrix with recombinant human bone morphogenetic protein-2 did not have pyrogen effect and the rabbits’ body temperature had no significant fluctuations after injection. The acute systemic toxicity test results showed that the nanometer human demineralized bone matrix with recombinant human bone morphogenetic protein-2 complied with the relevant provisions of the State, without obvious toxic reaction. The inflammatory response of nanometer human demineralized bone matrix with recombinant human bone morphogenetic protein-2 was significantly lighter than the reaction of β-tricalcium phosphate. The results showed that the nanometer human demineralized bone matrix with recombinant human bone morphogenetic protein-2 is a nanometer allogeneic bone graft substitutes with nontoxicity, good biocompatibility, high bioavailability, and less inflammatory reaction.