脱细胞真皮基质/磷酸氢钙骨复合支架的制备和体内成骨研究  

Preparation and in vivo osteogenesis of acellular dermal matrix/dicalcium phosphate composite scaffold for bone repair

作　　者：蓝益南 张静逸 冉永峰 李博 蔡晓斌 江涛 薛德挺[1] LAN Yinan;ZHANG Jingyi;RAN Yongfeng;LI Bo;CAI Xiaobin;JIANG Tao;XUE Deting(Department of Orthopedics,the Second Affiliated Hospital,College of Medicine,Zhejiang University,Hangzhou Zhejiang,310009,P.R.China;Department of Orthopedics,Lishui Hospital of Zhejiang University,Lishui Zhejiang,323000,P.R.China;Hangzhou Huamai Medical Technology Co.Ltd.,Hangzhou Zhejiang,310052,P.R.China)

机构地区：[1]浙江大学医学院附属第二医院骨科,杭州310009 [2]浙江大学丽水医院骨科,浙江丽水323000 [3]杭州华迈医疗科技有限公司,杭州310052

出　　处：《中国修复重建外科杂志》2024年第6期755-762,共8页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家重点研发计划项目(2019YFA0110600);浙江省医药卫生科技计划项目(2024KY1850、2024KY1851);浙江省科技厅项目(2017C37181)。

摘　　要：目的探讨脱细胞真皮基质(acellular dermal matrix,ADM)/磷酸氢钙(dicalcium phosphate,DCP)复合支架的理化特性、成骨性能及在兔股骨髁缺损模型中的成骨能力。方法采用微纤维化技术制备ADM/DCP复合支架,行DNA残留、脂肪含量及α-半乳糖基抗原(α-1,3-galactosyle,α-Gal)抗原表位数检测ADM/DCP复合支架的脱细胞效果;场发射扫描电镜观察及压汞仪测定支架孔隙率行支架微观结构表征;行支架X射线衍射分析晶型是否发生变化;支架溶解性检测溶液pH值和钙离子含量;支架体外矿化实验观察表面矿化物。选取健康雄性新西兰大白兔12只,制备兔股骨髁缺损模型,左、右侧缺损处分别植入ADM/DCP复合支架(实验组)和骼金?人工骨修复材料(对照组)。术后6、12周取材行大体观察;行Micro-CT检测并定量分析相关指标[骨体积(bone volume,BV)、骨体积分数(bone volume/tissue volume,BV/TV)、骨表面积和骨体积之比(bone surface/bone volume,BS/BV)、骨小梁厚度(trabecular thickness,Tb.Th)、骨小梁数目(trabecular number,Tb.N)、骨小梁间隙(trabecular separation,Tb.Sp)、骨密度(bone mineral density,BMD)];行HE染色和Masson染色,观察骨缺损修复及骨基质成熟情况。结果大体观察示ADM/DCP复合支架为白色海绵状固体。与ADM相比,ADM/DCP复合支架的DNA残留、脂肪含量和α-Gal抗原含量均大幅降低(P<0.05)。场发射扫描电镜观察示ADM/DCP复合支架具有多孔联通结构,猪真皮纤维上附着DCP颗粒;压汞仪测得ADM/DCP复合支架孔隙率为76.32%±1.63%。X射线衍射分析示,ADM/DCP复合支架中的DCP晶相保持完整。体外矿化结果示ADM/DCP复合支架的羟基磷灰石层基本成熟。兔股骨髁缺损修复实验示,术后12周切口完全愈合,无骨痂、骨赘。术后12周,Micro-CT观察示两组材料缺损部位已完成骨性愈合且生成大量新生骨组织,密度与周围骨组织无差异,两组材料成骨性能相当;除实验组Tb.Sp显�Objective To investigate the physicochemical properties,osteogenic properties,and osteogenic ability in rabbit model of femoral condylar defect of acellular dermal matrix(ADM)/dicalcium phosphate(DCP)composite scaffold.Methods ADM/DCP composite scaffolds were prepared by microfibril technique,and the acellular effect of ADM/DCP composite scaffolds was detected by DNA residue,fat content,andα-1,3-galactosyle(α-Gal)epitopes;the microstructure of scaffolds was characterized by field emission scanning electron microscopy and mercury porosimetry;X-ray diffraction was used to analyze the change of crystal form of scaffold;the solubility of scaffolds was used to detect the pH value and calcium ion content of the solution;the mineralization experiment in vitro was used to observe the surface mineralization.Twelve healthy male New Zealand white rabbits were selected to prepare the femoral condylar defect models,and the left and right defects were implanted with ADM/DCP composite scaffold(experimental group)and skeletal gold®artificial bone repair material(control group),respectively.Gross observation was performed at 6 and 12 weeks after operation;Micro-CT was used to detect and quantitatively analyze the related indicators[bone volume(BV),bone volume/tissue volume(BV/TV),bone surface/bone volume(BS/BV),trabecular thickness(Tb.Th),trabecular number(Tb.N),trabecular separation(Tb.Sp),bone mineral density(BMD)],and HE staining and Masson staining were performed to observe the repair of bone defects and the maturation of bone matrix.Results Gross observation showed that the ADM/DCP composite scaffold was a white spongy solid.Compared with ADM,ADM/DCP composite scaffolds showed a significant decrease in DNA residue,fat content,andα-Gal antigen content(P<0.05).Field emission scanning electron microscopy showed that the ADM/DCP composite scaffold had a porous structure,and DCP particles were attached to the porcine dermal fibers.The porosity of the ADM/DCP composite scaffold was 76.32%±1.63%measured by mercury porosimetry.

关 键 词：骨组织工程 脱细胞真皮基质 磷酸氢钙 复合支架 骨修复 

分 类 号：R318.08[医药卫生—生物医学工程]