Gong Weiyu, Huang Zhiwei, Dong Yanmei, Gan Yehua, Li Shenglin, Gao Xuejun, Chen Xiaofeng
Department of Cariology and Endodontology, Peking University School and Hospital of Stomatology, Beijing, China.
Department of Cariology and Endodontology, Peking University School and Hospital of Stomatology, Beijing, China.
J Endod. 2014 Jan;40(1):83-8. doi: 10.1016/j.joen.2013.08.018. Epub 2013 Nov 7.
This study aimed to investigate the effects of a novel nano-sized 58S bioactive glass (nano-58S BG) on the odontogenic differentiation and mineralization of human dental pulp cells (hDPCs) in vitro.
Extractions were prepared by incubating nano-58S BG, 45S5 BG, or 58S BG particulates in Dulbecco modified Eagle medium at 1% w/v for 24 hours and were filtrated through 0.22-μm filters. The supernatants were used as BG extractions. The hDPCs were cultured in nano-58S BG, 45S5 BG, and 58S BG extractions. The proliferation of hDPCs was evaluated using the methylthiazol tetrazolium assay. Odontogenic differentiation was evaluated based on the real-time polymerase chain reaction of differentiation- and mineralization-related genes, namely, alkaline phosphatase (ALP), collagen type I, dentin sialophosphoprotein (DSPP), and dentin matrix protein 1. The gene expressions were verified using ALP activity assessment, immunocytochemistry staining of osteocalcin and DSPP, and mineralization assay using alizarin red S stain.
All BG extractions up-regulated the expression of odontogenic genes, and the most significant enhancement was in the nano-58S BG group. All BG extractions, especially nano-58S, increased ALP activity, osteocalcin and DSPP protein production, and mineralized nodules formation.
Compared with regular BG, the novel nano-58S BG can induce the differentiation and mineralization of hDPCs more efficiently and might be a better potential candidate for dentin-pulp complex regeneration.
本研究旨在探讨新型纳米级58S生物活性玻璃(纳米-58S BG)对人牙髓细胞(hDPCs)体外牙源性分化和矿化的影响。
将纳米-58S BG、45S5 BG或58S BG颗粒在杜氏改良 Eagle培养基中以1% w/v的浓度孵育24小时,制备提取物,然后通过0.22μm滤器过滤。上清液用作BG提取物。将hDPCs培养在纳米-58S BG、45S5 BG和58S BG提取物中。使用甲基噻唑四氮唑法评估hDPCs的增殖。基于分化和矿化相关基因,即碱性磷酸酶(ALP)、I型胶原蛋白、牙本质涎磷蛋白(DSPP)和牙本质基质蛋白1的实时聚合酶链反应,评估牙源性分化。通过ALP活性评估、骨钙素和DSPP的免疫细胞化学染色以及茜素红S染色的矿化试验验证基因表达。
所有BG提取物均上调牙源性基因的表达,纳米-58S BG组的增强最为显著。所有BG提取物,尤其是纳米-58S,均增加了ALP活性、骨钙素和DSPP蛋白的产生以及矿化结节的形成。
与常规BG相比,新型纳米-58S BG能更有效地诱导hDPCs的分化和矿化,可能是牙本质-牙髓复合体再生的更好潜在候选物。