Department of Anatomy, Histology and Embryology, Faculty of Medicine, University of Debrecen, Nagyerdei krt 98, Debrecen, 4032, Hungary.
Department of Biomedical Sciences, Humanitas University, Via Rita Levi Montalcini 4, Rozzano, Milan, 20090, Italy; Humanitas Clinical and Research Center, Via Alessandro Manzoni 56, Rozzano, Milan, 20089, Italy.
Differentiation. 2019 May-Jun;107:24-34. doi: 10.1016/j.diff.2019.05.002. Epub 2019 May 22.
Bone graft substitutes and bone void fillers are predominantly used to treat bone defects and bone fusion in orthopaedic surgery. Some aragonite-based scaffolds of coralline exoskeleton origin exhibit osteoconductive properties and are described as useful bone repair scaffolds. The purpose of this study was to evaluate the in vitro osteogenic potential of the bone phase of a novel aragonite-based bi-phasic osteochondral scaffold (Agili-C™, CartiHeal Ltd.) using adult human bone marrow-derived mesenchymal stem cells (MSCs). Analyses were performed at several time intervals: 3, 7, 14, 21, 28 and 42 days post-seeding. Osteogenic differentiation was assessed by morphological characterisation using light microscopy after Alizarin red and von Kossa staining, and scanning electron microscopy. The transcript levels of alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), bone gamma-carboxyglutamate (BGLAP), osteonectin (SPARC) and osteopontin (SPP1) were determined by quantitative PCR. Proliferation was assessed by a thymidine incorporation assay and proliferating cell nuclear antigen (PCNA) immunocytochemistry. Our results demonstrate that the bone phase of the bi-phasic aragonite-based scaffold supports osteogenic differentiation and enhanced proliferation of bone marrow-derived MSCs at both the molecular and histological levels. The scaffold was colonized by differentiating MSCs, suggesting its suitability for incorporation into bone voids to accelerate bone healing, remodelling and regeneration. The mechanism of osteogenic differentiation involves scaffold surface modification with de novo production of calcium phosphate deposits, as revealed by energy dispersive spectroscopy (EDS) analyses. This novel coral-based scaffold may promote the rapid formation of high quality bone during the repair of osteochondral lesions.
骨移植替代物和骨空洞填充剂主要用于治疗骨科手术中的骨缺损和骨融合。一些源自珊瑚外骨骼的方解石基支架具有骨诱导特性，被描述为有用的骨修复支架。本研究旨在使用成人骨髓间充质干细胞（MSCs）评估新型方解石基双相骨软骨支架（Agili-C™，CartiHeal Ltd.）的骨相的体外成骨潜力。分析在几个时间间隔进行：接种后 3、7、14、21、28 和 42 天。通过茜素红和 von Kossa 染色后使用光学显微镜进行形态学特征分析以及扫描电子显微镜来评估成骨分化。通过定量 PCR 测定碱性磷酸酶（ALP）、 runt 相关转录因子 2（RUNX2）、骨γ-羧基谷氨酸（BGLAP）、骨粘连蛋白（SPARC）和骨桥蛋白（SPP1）的转录水平。通过胸苷掺入测定和增殖细胞核抗原（PCNA）免疫细胞化学评估增殖。我们的结果表明，双相方解石基支架的骨相支持骨髓间充质干细胞的成骨分化和增强增殖，在分子和组织学水平上均如此。支架被分化的 MSC 定植，表明其适合掺入骨空洞以加速骨愈合、重塑和再生。成骨分化的机制涉及支架表面改性，通过能量色散光谱（EDS）分析揭示了新生成的磷酸钙沉积物。这种新型珊瑚基支架可能会在修复骨软骨病变时促进高质量骨的快速形成。
