Department of Orthopedics, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu Province, People's Republic of China.
Department of Trauma Center, The Affiliated Changzhou No.2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu Province, People's Republic of China.
Int J Nanomedicine. 2023 Oct 21;18:5907-5923. doi: 10.2147/IJN.S428965. eCollection 2023.
This study aims to investigate the impact of enhancing subchondral bone repair on the efficacy of articular cartilage restoration, thereby achieving improved osteochondral regeneration outcomes.
In this study, we modified the surface of nano-hydroxyapatite (n-HAp) through alkylation reactions to prepare n-HApMA. Characterization techniques, including X-ray diffraction, infrared spectroscopy scanning, thermogravimetric analysis, particle size analysis, and electron microscopy, were employed to analyze n-HApMA. Bioinks were prepared using n-HApMA, high porosity GelMA hydrogel, and adipose tissue derived stromal cells (ADSCs). The rheological properties of the bioinks during photocuring were investigated using a rheometer. Based on these bioinks, a biphasic scaffold was constructed. The viability of cells within the scaffold was observed using live-dead cell staining, while the internal morphology was examined using scanning electron microscopy. The stiffness of the scaffold was evaluated through compression testing. Scaffolds were implanted into the osteochondral defects of New Zealand rabbit knees, and microCT was utilized to observe the subchondral bone repair. Hematoxylin and eosin (H&E) staining, Masson's trichrome staining, and Safranin O/Fast Green staining were performed to assess the regeneration of subchondral bone and cartilage. Furthermore, immunohistochemical staining was employed to detect the expression of osteogenic and chondrogenic-related molecules.
Scaffold characterization revealed that surface modification enables the uniform distribution of n-HApMA within the GelMA matrix. The incorporation of 5% n-HApMA notably enhanced the elastic modulus and stiffness of the 6% high-porosity GelMA in comparison to n-HAp. Moreover, in-vivo study showed that the homogeneous dispersion of n-HApMA on the GelMA matrix facilitated the osteogenic differentiation of adipose-derived stem cells (ADSCs) and promoted osteochondral tissue regeneration.
These findings suggest potential applications of the n-HApMA/GelMA composite in the field of tissue engineering and regenerative medicine.
本研究旨在探讨增强软骨下骨修复对关节软骨修复效果的影响,从而实现更好的骨软骨再生效果。
本研究通过烷基化反应对纳米羟基磷灰石(n-HAp)进行表面修饰,制备 n-HApMA。采用 X 射线衍射、红外光谱扫描、热重分析、粒度分析和电子显微镜等表征技术对 n-HApMA 进行分析。采用 n-HApMA、高孔隙率 GelMA 水凝胶和脂肪组织来源的基质细胞(ADSCs)制备生物墨水。使用流变仪研究生物墨水在光固化过程中的流变性能。基于这些生物墨水构建了双相支架。使用活死细胞染色观察细胞在支架内的活力,使用扫描电子显微镜观察内部形态。通过压缩试验评估支架的硬度。将支架植入新西兰兔膝关节的骨软骨缺损部位,利用 microCT 观察软骨下骨修复情况。进行苏木精和伊红(H&E)染色、Masson 三色染色和番红 O/快绿染色,评估软骨下骨和软骨的再生情况。此外,还进行了免疫组织化学染色,检测成骨和软骨形成相关分子的表达。
支架表征结果表明,表面修饰使 n-HApMA 在 GelMA 基质中均匀分布。与 n-HAp 相比,5% n-HApMA 的掺入可显著提高 6%高孔隙率 GelMA 的弹性模量和硬度。此外,体内研究表明,n-HApMA 在 GelMA 基质中的均匀分散促进了脂肪来源干细胞(ADSCs)的成骨分化,促进了骨软骨组织的再生。
这些发现表明 n-HApMA/GelMA 复合材料在组织工程和再生医学领域具有潜在应用价值。