两种去抗原异种骨性能评价及修复大鼠颅骨缺损实验研究  

Performance evaluation of two antigen-extracted xenogeneic ostein and experimental study on repairing skull defects in rats

作　　者：李矛 白玉龙 李淼 周建伟[1] LI Mao;BAI Yulong;LI Miao;ZHOU Jianwei(Department of Orthopedics,Beijing Tongren Hospital,Capital Medical University,Beijing,100730,P.R.China;Shanghai Gencong Biomedical Materials Research Center,Shanghai,201201,P.R.China;Shanghai Yapeng Biotechnology Co.,Ltd,Shanghai,201201,P.R.China)

机构地区：[1]首都医科大学附属北京同仁医院骨科,北京100730 [2]上海亘从生物医用材料研究中心,上海201201 [3]上海亚朋生物技术有限公司,上海201201

出　　处：《中国修复重建外科杂志》2021年第10期1303-1310,共8页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的评价两种去抗原异种骨支架脱钙骨基质(decalcified bone matrix,DBM)和煅烧骨的理化性能、免疫原性及成骨能力。方法取成年猪股骨,通过除去异种骨无机成分和有机成分的方式,分别制备异种DBM和煅烧骨。测量并计算两种材料的密度、pH值,扫描电镜观察材料形貌并测量孔径,全自动接触角测量仪测量材料的表面接触角;并通过细胞毒性实验、成骨细胞增殖实验、DNA残留检测和人外周血淋巴细胞增殖实验评价两种材料的安全性、成骨活性及免疫原性。将两种材料分别植入6周龄雄性SD大鼠颅骨直径5 mm全层缺损(空白对照组不植入材料),术后4、8周取材,通过大体观察、Micro-CT扫描和HE染色观察评价材料对大鼠颅骨的修复效果。结果与煅烧骨比较,DBM密度较小,亲水性较差;两种材料pH值在5.5~6.1之间,孔径介于160~800μm之间。两种材料均无细胞毒性且能够促进成骨细胞增殖,培养1、4、7 d成骨细胞增殖吸光度(A)值DBM组均显著大于煅烧骨组(P<0.05)。两种材料DNA残留量远低于50 ng/mg干重,且均不会刺激人外周血淋巴细胞增殖和分化。动物体内实验结果显示,术后4周DBM组和煅烧骨组新生骨体积百分比(bone volume/total volume,BV/TV)均显著高于空白对照组,煅烧骨组显著高于DBM组,差异均有统计学意义(P<0.05);术后8周,3组间BV/TV差异无统计学意义(P>0.05)。HE染色示,术后4、8周,空白对照组缺损部位被纤维结缔组织填充,缺损明显,未见骨长入;DBM组和煅烧骨组缺损均已被不同程度修复,可见大量新骨生成,DBM组材料降解性优于煅烧骨组。结论两种异种骨支架理化性能及降解性略有差异,无免疫原性,且均能促进大鼠颅骨缺损的修复。Objective To evaluate the physical and chemical properties, immunogenicity, and osteogenesis of two antigen-extracted xenogeneic bone scaffolds—decalcified bone matrix(DBM) and calcined bone. Methods By removing the inorganic and organic components of adult pig femus, xenogeneic DBM and calcined bone were prepared respectively. The density and pH value of the two materials were measured and calculated, the material morphology and pore diameter were observed by scanning electron microscope, and the surface contact angle was measured by automatic contact angle measuring instrument. The safety, osteogenic activity, and immunogenicity of the two materials were evaluated by cytotoxicity test, osteoblast proliferation test, DNA residue test, and human peripheral blood lymphocyte proliferation test. The two materials were implanted into the 5 mm full-thickness skull defect of 6-week-old male Sprague Dawley rats(the blank control group was not implanted with materials). The materials were taken at 4 and 8 weeks after operation, the repair effect of the materials on the rat skull was observed and evaluated by gross observation, Micro-CT scanning, and HE staining observation. Results Compared with calcined bone, DBM has lower density and poor hydrophilicity;the pH value of the two materials was 5.5-6.1, and the pore diameter was 160-800 μm. The two materials were non-cytotoxic and could promote the proliferation of osteoblasts. The absorbance(A) values of osteoblast proliferation at 1, 4, and 7 days in the DBM group were significantly higher than those in the calcined bone group(P<0.05). The DNA residues of the two materials were much lower than 50 ng/mg dry weight, and neither of them could stimulate the proliferation and differentiation of human peripheral blood lymphocytes. The results of animal experiments in vivo showed that the bone volume/total volume(BV/TV) in DBM group and calcined bone group were significantly higher than that in blank control group at 4 weeks after operation(P<0.05), and that in calcined bone

关 键 词：异种骨 脱钙骨基质 煅烧骨 颅骨缺损 大鼠 

分 类 号：R318.08[医药卫生—生物医学工程]