Wang Zhifa, Han Leng, Sun Tianyu, Ma Junli, Sun Shuohui, Ma Limin, Wu Buling
School of Stomatology, Southern Medical University, Nanfang Hospital, Southern Medical University, Guangzhou 510515, PR China; Department of Stomatology, General Hospital of Southern Theater of PLA, Guangzhou 510010, China.
Department of Pathology, General Hospital of Southern Theater of PLA, Guangzhou 510010, China.
Acta Biomater. 2020 Dec;118:54-68. doi: 10.1016/j.actbio.2020.10.022. Epub 2020 Oct 15.
Bioactive scaffolds from synthetical polymers or decellularized cartilage matrices have been widely used in osteochondral regeneration. However, the risks of potential immunological reactions and the inevitable donor morbidity of these scaffolds have limited their practical applications. To address these issues, a biological extracellular matrix (ECM) scaffold derived from allogenic decellularized bone marrow mesenchymal stem cell (BMSC) sheets was established for osteochondral reconstruction. BMSCs were induced to form cell sheets. Three different concentrations of sodium dodecyl sulfate (SDS), namely, 0.5%, 1%, and 3%, were used to decellularize these BMSC sheets to prepare the ECM. Histological and microstructural observations were performed in vitro and then the ECM scaffolds were implanted into osteochondral defects in rabbits to evaluate the repair effect in vivo. Treatment with 0.5% SDS not only efficiently removed BMSCs but also successfully preserved the original structure and bioactive components of the ECM When compared with the 1% and 3% SDS groups, histological observations substantiated the superior repair effect of osteochondral defects, including the simultaneous regeneration of well-vascularized subchondral bone and avascular articular cartilage integrated with native tissues in the 0.5% SDS group. Moreover, RT-PCR indicated that ECM scaffolds could promote the osteogenic differentiation potential of BMSCs under osteogenic conditions while increasing the chondrogenic differentiation potential of BMSCs under chondrogenic conditions. Allogenic BMSC sheets decellularized with 0.5% SDS treatment increased the recruitment of BMSCs and significantly improved the regeneration of osteochondral defects in rabbits, thus providing a prospective approach for both articular cartilage and subchondral bone reconstruction with cell-free transplantation.
合成聚合物或脱细胞软骨基质制成的生物活性支架已广泛应用于骨软骨再生。然而,这些支架存在潜在免疫反应风险以及不可避免的供体发病问题,限制了它们的实际应用。为解决这些问题,建立了一种源自同种异体脱细胞骨髓间充质干细胞(BMSC)片层的生物细胞外基质(ECM)支架用于骨软骨重建。诱导BMSC形成细胞片层。使用三种不同浓度的十二烷基硫酸钠(SDS),即0.5%、1%和3%,对这些BMSC片层进行脱细胞处理以制备ECM。在体外进行组织学和微观结构观察,然后将ECM支架植入兔的骨软骨缺损处,以评估体内修复效果。0.5% SDS处理不仅有效去除了BMSC,还成功保留了ECM的原始结构和生物活性成分。与1%和3% SDS组相比,组织学观察证实了0.5% SDS组对骨软骨缺损的修复效果更佳,包括同时再生血管丰富的软骨下骨和与天然组织整合的无血管关节软骨。此外,RT-PCR表明,ECM支架在成骨条件下可促进BMSC的成骨分化潜能,同时在软骨形成条件下增加BMSC的软骨形成分化潜能。经0.5% SDS处理脱细胞的同种异体BMSC片层增加了BMSC的募集,并显著改善了兔骨软骨缺损的再生,从而为无细胞移植的关节软骨和软骨下骨重建提供了一种前瞻性方法。