Department of Oral Biology, Faculty of Dentistry, Mahidol University, 6 Yothi Street, Ratchathewi, Bangkok, Thailand.
Department of Implantology, School and Hospital of Stomatology, Kunming Medical University, Hecheng International Community, Building C, No.1088 the Middle of Hai Yuan Road, Wuhua District, Kunming, Yunnan, People's Republic of China.
Sci Rep. 2023 Aug 12;13(1):13140. doi: 10.1038/s41598-023-40258-7.
Demineralized dentin matrix (DDM) is an osteoconductive and osteoinductive material that has been successfully used in sinus floor augmentation and alveolar ridge augmentation in clinical applications. It releases bone morphogenetic proteins (BMPs) and other growth factors, making DDM a suitable grafting material. However, the granular particle of DDM makes it difficult to anchor into the bone defect area. The aim of this study was to investigate the biological effects and osteoinductivity of the combination of DDM and Fibrin Glue (FG) at an optimal ratio on bone healing from a critical bone defect in an animal model. The mouse osteoblastic cell line (MC3T3-E1) was co-cultured with various ratios of DDM and FG to examine their effects on osteoblast proliferation and differentiation, as indicated by alkaline phosphatase (ALP) activity, osteocalcin (OC) production and mineralized nodules formation. The optimal ratio was then chosen for further study with a rabbit calvarial defective model, in which they were implanted with DDM or DDM-FG1 (1 g: 0.1 ml) and DDM-FG2 (1 g: 0.5 ml) compounds, or left blank for 2, 4, 8 and 12 weeks to investigate soft tissue and new bone regeneration. Micro-CT and histology analysis were used to evaluate the total grafting properties according to the different healing periods. The result from in vitro studies demonstrated that the ratio of 1:0.1 induced more ALP activity and mineralized nodules, while the ratio of 1: 0.5 (DDM-FG combined) induced more osteocalcin (OC) at specific time points. In the animal model, the 3D new bone volume in all DDM-FG treatment groups was significantly greater than that in the blank group at 2, 4, 8 and 12 weeks. Furthermore, the new bone volume was greater in DDM-FG2 when compared to the other groups during the early weeks of the healing period. In histological analysis, clusters of osteoblasts were formed adjacent to the DDM particles, and newly formed bone was observed in all groups, suggesting an osteoinductive property of DDM. Moreover, the greater new collagen synthesis observed at 4 weeks suggested that early bone healing was induced in the DDM-FG2 group. This study demonstrated that at an optimal ratio, the DDM-FG compound enhances osteogenic activities and bone regeneration.
脱矿牙本质基质 (DDM) 是一种具有骨诱导和骨传导作用的材料，已成功应用于临床中的鼻窦底提升和牙槽嵴增高。它释放骨形态发生蛋白 (BMPs) 和其他生长因子，使 DDM 成为一种合适的移植物材料。然而，DDM 的颗粒状使得其难以固定在骨缺损区域。本研究旨在探讨在动物模型中，从临界骨缺损处的骨愈合角度出发，DDM 与纤维蛋白胶 (FG) 以最佳比例结合的生物学效应和骨诱导性。将小鼠成骨细胞系 (MC3T3-E1) 与不同比例的 DDM 和 FG 共培养，通过碱性磷酸酶 (ALP) 活性、骨钙素 (OC) 产生和矿化结节形成来检测其对成骨细胞增殖和分化的影响。然后，选择最佳比例的 DDM 和 FG 用于进一步研究，将 DDM 或 DDM-FG1(1g:0.1ml)和 DDM-FG2(1g:0.5ml) 复合物植入兔颅骨缺损模型中，并在 2、4、8 和 12 周时观察空白组，以研究软组织和新骨再生。使用微 CT 和组织学分析根据不同的愈合期评估总移植物特性。体外研究结果表明，1:0.1 的比例诱导更多的 ALP 活性和矿化结节，而 1:0.5(DDM-FG 联合)的比例在特定时间点诱导更多的骨钙素 (OC)。在动物模型中，所有 DDM-FG 治疗组的 3D 新骨体积在 2、4、8 和 12 周时均明显大于空白组。此外，在愈合早期，DDM-FG2 组的新骨体积大于其他组。在组织学分析中，成骨细胞簇形成于 DDM 颗粒附近，所有组均观察到新骨形成，提示 DDM 具有骨诱导性。此外，在 4 周时观察到更多的新胶原合成，提示 DDM-FG2 组诱导了早期骨愈合。本研究表明，在最佳比例下，DDM-FG 复合物增强了成骨活性和骨再生。
