Shi PeiKai, Zhou WanTong, Dong JingBo, Li ShuJun, Lv PengJun, Liu Chenxi
Department of Oral Implantology, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang, 050017, PR China.
Heliyon. 2022 Jan 19;8(1):e08796. doi: 10.1016/j.heliyon.2022.e08796. eCollection 2022 Jan.
The study aimed to investigate the osteogenic ability of bioactive glass (bioglass) combined with recombinant human bone morphogenetic protein-9 (rhBMP-9) on rat bone marrow mesenchymal stem cells (BMSCs) in vitro. The study also compares bone regeneration using rhBMP9 soaked with different carrier systems, including bioglass or collagen membranes (BioGide, BG) in a rat alveolar bone site preservation model in vivo.
Scanning electron microscopy was employed to analyze bioglass surface. The absorption and release potential of rhBMP9 from bioglass were researched by ELISA.The cell viability, adhesion, proliferation, and differentiation were assessed for rhBMP9 soaked on bioglass by cck-8 kit, alkaline phosphatase (ALP) activity assay, alizarin red staining, and real-time PCR. Furthermore, prepared grafts (bioglass + BG, bioglass/rhBMP9+BG, and bioglass + BG/rhBMP9) were implanted into the maxillary right first incisor sockets of Sprague Dawley rats for 8 weeks, and new bone formation was quantified by micro-CT and histological analysis.
Bioglass absorbed rhBMP9 dramatically and released it with a slow and stable speed within ten days by ELISA. When used with cck-8 kit detection, cell viability at 24 h, cell adhesion rate at 8 h, and cell proliferation at 1, 3, and 5 days were decreased in the bioglass alone group versus the control group but slightly increased with the addition of rhBMP9. Similarly, the effect of osteogenic differentiation on bioglass increased significantly when combined with rhBMP9 by upregulating the expression of ALP, mineralized matrix, and osteogenic related genes. Furthermore, both bioglass/rhBMP9+BG samples and bioglass + BG/rhBMP9 samples significantly improved several bone formation parameters compared with bioglass + BG samples. Interestingly, bioglass + BG/rhBMP9 samples demonstrated more bone regeneration in rat site preservation models.
Both bioglass and BG can be applied in GBR surgery as effective carriers of rhBMP9. However, BG may be more suitable than bioglass for investigating site preservation effect after tooth extraction when associated with rhBMP9 and provides a practical clinical solution to the problem of bone deficiency caused by alveolar bone atrophy.
本研究旨在体外研究生物活性玻璃（生物玻璃）联合重组人骨形态发生蛋白-9（rhBMP-9）对大鼠骨髓间充质干细胞（BMSCs）的成骨能力。该研究还在大鼠牙槽骨位点保存模型体内比较了使用不同载体系统（包括生物玻璃或胶原膜（BioGide，BG））浸泡rhBMP9后的骨再生情况。
采用扫描电子显微镜分析生物玻璃表面。通过酶联免疫吸附测定法研究rhBMP9从生物玻璃中的吸收和释放潜力。通过cck-8试剂盒、碱性磷酸酶（ALP）活性测定、茜素红染色和实时聚合酶链反应评估浸泡在生物玻璃上的rhBMP9对细胞活力、粘附、增殖和分化的影响。此外，将制备好的移植物（生物玻璃+BG、生物玻璃/rhBMP9+BG和生物玻璃+BG/rhBMP9）植入Sprague Dawley大鼠右上颌第一切牙牙槽窝8周，通过显微计算机断层扫描和组织学分析对新骨形成进行定量。
酶联免疫吸附测定法显示生物玻璃能大量吸收rhBMP9，并在十天内以缓慢且稳定的速度释放。当使用cck-8试剂盒检测时，与对照组相比，单独生物玻璃组在24小时的细胞活力、8小时的细胞粘附率以及1、3和5天的细胞增殖均降低，但添加rhBMP9后略有增加。同样，与rhBMP9联合使用时，生物玻璃上的成骨分化作用通过上调ALP、矿化基质和成骨相关基因的表达而显著增强。此外，与生物玻璃+BG样本相比，生物玻璃/rhBMP9+BG样本和生物玻璃+BG/rhBMP9样本均显著改善了多个骨形成参数。有趣的是，在大鼠位点保存模型中，生物玻璃+BG/rhBMP9样本显示出更多的骨再生。
生物玻璃和BG均可作为rhBMP9的有效载体应用于引导骨再生手术。然而，在与rhBMP9联合使用时，BG可能比生物玻璃更适合研究拔牙后的位点保存效果，并为牙槽骨萎缩引起的骨缺损问题提供切实可行的临床解决方案。
