异种脱钙骨基质颗粒的生物相容性实验研究  

EXPERIMENTAL STUDIES ON BIOCOMPATIBILITY OF HETEROGENEOUS DEMINERALIZED BONE MATRIX PARTICLES

作　　者：梁新刚[1,2] 杨志明[1] 项舟[1,2] 李秀群[1] 黄光平[1,2] 智伟[1] 

机构地区：[1]四川大学华西医院生物治疗国家重点实验室.干细胞与组织工程研究室,成都610041 [2]四川大学华西医院骨科

出　　处：《中国修复重建外科杂志》2009年第1期76-81,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(30271317;30570469)~~

摘　　要：目的评价制备的异种脱钙骨基质(demineralized bone matrix,DBM)植入体内的生物相容性,为进一步实验研究和临床应用提供依据。方法取猪四肢长管状骨皮质骨经理化处理后,制备脱脂脱钙(材料A)和脱脂脱钙去细胞(材料B)两种粒径为250～810μm的DBM颗粒和浸提液,骨颗粒脱脂脱钙后测钙、磷含量。采用标准的毒理学方法进行急性毒性实验、皮肤刺激实验、热原实验、溶血实验、细胞毒性实验和肌肉植入实验。结果未脱钙皮质骨钙、磷含量分别为(189.09±3.12)mg/g和(124.73±2.87)mg/g;DBM颗粒的钙、磷含量分别为(3.48±0.09)mg/g和(3.46±0.07)mg/g;DBM颗粒钙、磷含量分别为未脱钙皮质骨的1.87%和2.69%。急性毒性实验:各组小鼠活动正常,7d内小鼠无死亡,各组动物未见中毒症状或不良反应;7d后各组小鼠体重日平均增加差异无统计学意义(P>0.05)。皮内刺激实验观察显示,注射材料A、B浸提液和生理盐水处无明显红斑、水肿和皮肤坏死,极轻微刺激。热原实验两组动物体温升高最高均为0.4℃,符合<0.6℃的国家标准。材料A浸提液的溶血率为1.14%,材料B为0.93%,符合<5%的国家标准。异种DBM的细胞毒性为0～1级。肌肉内植入实验显示动物术后生存良好,各植入部位均未见组织坏死、积液及化脓感染。术后不同时间点植入材料A、B组局部细胞免疫定量评分比较差异无统计学意义(P>0.05)。结论DBM颗粒的毒性程度为无毒,皮肤无刺激,无热原性,溶血率<5%,对细胞无明显毒性,具有良好的生物相容性和一定的骨诱导性。目的评价制备的异种脱钙骨基质(demineralizedbonematrix,DBM)植入体内的生物相容性,为进一步实验研究和临床应用提供依据。方法取猪四肢长管状骨皮质骨经理化处理后,制备脱脂脱钙(材料A)和脱脂脱钙去细胞(材料B)两种粒径为250～810μm的DBM颗粒和浸提液,骨颗粒脱脂脱钙后测钙、磷含量。采用标准Objective To evaluate the biocompatibility of manufactured heterogeneous demineralized bone matrix （DBM） particles and to provide basis for further experimental study and clinical application. Methods Heterogeneous DBM particles A （degreased and demineralized） and B （degreased, demineralized and acellular）, particle size from 250 to 810 tam, and leaching liquor were made with a series of physical and chemical methods from pig limbs cortical bone. The residual calcium and phosphorus contents of bone particles were measured after degreased and demineralized. The acute toxicity test, skin stimulating test, pyrogeneous test, hemolysis test, cellular toxicity test and muscular embedded test were carried out according standard toxicological method. Results The contents of calcium and phosphorus in cortical bone were （189.09 ± 3.12） mg/g and （124.73 ± 2.87） mg/g, and in demineralized bone matrix particles were （3.48 ± 0.09） mg/g and （3.46 ± 0.07） mg/g. The residual calcium content was 1.87%, of phosphorus was 2.69%. The activity of mice was normal in the acute toxicity test. No animal died and no toxicity symptom or adverse effects were shown within 7 days. The mean weight daily increased showed no statistically significant difference （P 〉 0.05） between two groups after 7 days. Skin stimulating reactions were not found in the two experimental groups and negative control group by intradermal stimulation test. The maximal increase of body temperature in two experimental groups were 0.4 ℃ , which meet the national standard （〈 0.6℃）. The rate of haemolysis to the leaching liquor was 1.14% （A） and 0.93% （B）, which was lower than the national standard （〈 5%）. The cell proliferation rates of two experimental groups when compared with control group showed no statistically significant difference （P 〉 0.05）. The toxicity of DBM particles leaching liquor was graded from 0 to l, which means the material has no cytotoxicity. All the animals survived well. There was

关 键 词：脱钙骨基质 异种骨 生物相容性 

分 类 号：R318.08[医药卫生—生物医学工程] R318[医药卫生—基础医学]