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  • Title: [Preparation and in vivo osteogenesis of acellular dermal matrix/dicalcium phosphate composite scaffold for bone repair].
    Author: Lan Y, Zhang J, Ran Y, Li B, Cai X, Jiang T, Xue D.
    Journal: Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi; 2024 Jun 15; 38(6):755-762. PubMed ID: 38918199.
    Abstract:
    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, and α-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, and α-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. X-ray diffraction analysis showed that the crystalline phase of DCP in the ADM/DCP composite scaffolds remained intact. Mineralization results in vitro showed that the hydroxyapatite layer of ADM/DCP composite scaffolds was basically mature. The repair experiment of rabbit femoral condyle defect showed that the incision healed completely after operation without callus or osteophyte. Micro-CT showed that bone healing was complete and a large amount of new bone tissue was generated in the defect site of the two groups, and there was no difference in density between the defect site and the surrounding bone tissue, and the osteogenic properties of the two groups were equivalent. There was no significant difference in BV, BV/TV, BS/BV, Tb.Th, Tb.N, and BMD between the two groups ( P>0.05), except that the Tb.Sp in the experimental group was significantly higher than that in the control group ( P<0.05). At 6 and 12 weeks after operation, HE staining and Masson staining showed that the new bone and autogenous bone fused well in both groups, and the bone tissue tended to be mature. CONCLUSION: The ADM/DCP composite scaffold has good biocompatibility and osteogenic ability similar to the artificial bone material in repairing rabbit femoral condylar defects. It is a new scaffold material with potential in the field of bone repair. 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 显著大于对照组（ P<0.05）外，两组BV、BV/TV、BS/BV、Tb.Th、Tb.N、BMD比较差异均无统计学意义（ P>0.05）。术后6、12周HE染色和Masson染色示两组新生骨和自体骨融合良好，骨组织均趋于成熟。. 结论: ADM/DCP复合支架具有良好生物相容性，修复兔股骨髁缺损具有与人工骨修复材料相似的成骨能力，是骨修复领域具有应用潜力的新型支架材料。.
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