Department of Removable Prosthodontics, School of Dental Medicine, Tsurumi University, 2-1-3 Tsurumi, Tsurumi-Ku, Yokohama, 230-8501, Japan.
Department of Biochemistry and Molecular Biology, School of Dental Medicine, Tsurumi University, 2-1-3 Tsurumi, Tsurumi-Ku, Yokohama, 230-8501, Japan.
Int J Implant Dent. 2022 Nov 1;8(1):49. doi: 10.1186/s40729-022-00449-9.
To investigate the bone augmentation ability of demineralized bone sheets mixed with allogeneic bone with protein fractions containing bioactive substances and the interaction between coexisting bioactive substances and proteins.
Four types of demineralized bone sheets mixed with allogeneic bone in the presence or absence of bone proteins were created. Transplantation experiments using each demineralized bone sheet were performed in rats, and their ability to induce bone augmentation was analysed by microcomputed tomography images. Bioactive substances in bone proteins were isolated by heparin affinity chromatography and detected by the measurement of alkaline phosphatase activity in human periodontal ligament cells and dual luciferase assays. Noncollagenous proteins (NCPs) coexisting with the bioactive substances were identified by mass spectrometry, and their interaction with bioactive substances was investigated by in vitro binding experiments.
Demineralized bone sheets containing bone proteins possessed the ability to induce bone augmentation. Bone proteins were isolated into five fractions by heparin affinity chromatography, and transforming growth factor-beta (TGF-β) was detected in the third fraction (Hep-c). Dentin matrix protein 1 (DMP1), matrix extracellular phosphoglycoprotein (MEPE), and biglycan (BGN) also coexisted in Hep-c, and the binding of these proteins to TGF-β increased TGF-β activity by approximately 14.7% to 32.7%.
Demineralized bone sheets are capable of inducing bone augmentation, and this ability is mainly due to TGF-β in the bone protein mixed with the sheets. The activity of TGF-β is maintained when binding to bone NCPs such as DMP1, MEPE, and BGN in the sheets.
研究脱矿骨片与含有生物活性物质的同种异体骨蛋白混合的骨增强能力，以及共存生物活性物质与蛋白质之间的相互作用。
制备了四种含有或不含有骨蛋白的脱矿骨片与同种异体骨混合的脱矿骨片。在大鼠中进行了使用每种脱矿骨片的移植实验，通过 microCT 图像分析其诱导骨增强的能力。通过肝素亲和层析分离骨蛋白中的生物活性物质，并通过测量人牙周膜细胞的碱性磷酸酶活性和双荧光素酶测定法检测其活性。通过质谱鉴定与生物活性物质共存的非胶原蛋白（NCPs），并通过体外结合实验研究它们与生物活性物质的相互作用。
含有骨蛋白的脱矿骨片具有诱导骨增强的能力。肝素亲和层析将骨蛋白分离成五个部分，在第三部分（Hep-c）中检测到转化生长因子-β（TGF-β）。牙本质基质蛋白 1（DMP1）、基质细胞外磷酸糖蛋白（MEPE）和 biglycan（BGN）也共存于 Hep-c 中，这些蛋白质与 TGF-β 的结合使 TGF-β 的活性增加了约 14.7%至 32.7%。
脱矿骨片具有诱导骨增强的能力，这种能力主要归因于与片混合的骨蛋白中的 TGF-β。当与 DMP1、MEPE 和 BGN 等片内的骨 NCPs 结合时，TGF-β 的活性得以维持。
