Ohlrich E J, Coates D E, Cullinan M P, Milne T J, Zafar S, Zhao Y, Duncan W D, Seymour G J
Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, PO Box 647, Dunedin 9054, New Zealand.
Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, PO Box 647, Dunedin 9054, New Zealand.
Arch Oral Biol. 2016 Mar;63:7-14. doi: 10.1016/j.archoralbio.2015.11.013. Epub 2015 Nov 23.
Osteonecrosis of the jaws is recognised as a serious complication for patients receiving bisphosphonates. The anti-angiogenic effects of bisphosphonates have been implicated in the pathogenesis of bisphosphonate-related osteonecrosis of the jaw (BRONJ). The purpose of this study was to determine the effects of zoledronic acid on cultured human gingival fibroblasts in relation to the modulation of genes associated with angiogenic regulation.
Primary cultures of fibroblasts were developed from gingival tissues excised during crown-lengthening surgery from three patients. Cells were cultured with and without 30μM zoledronic acid for 6, 12 and 24h and cellular proliferation and migration investigated using CellTiter-Blue and scratch wound assays, respectively. Gene expression was determined using semi-quantitative PCR array technology that allowed the analysis of 84 pathway-focused genes known to be important in the regulation of angiogenesis.
Zoledronic acid increased the proliferation of the gingival fibroblasts in a dose dependent manner with 12 and 24h of exposure. Scratch wounding of the human gingival fibroblasts and treatment with increasing doses and time exposure to zoledronic acid (ZA) inhibited their migration. Statistically significant increases in gene expression were found for RHOB, VEGFA, CD55 and BMP2 (p≤0.05) in response to 30μM zoledronic acid. CCL2 and IL6 genes were significantly downregulated (p≤0.05).
The regulation of the prenylated protein RHOB in this study was consistent with the known effects of zoledronic acid on the mevalonate pathway. The down regulation of CCL2 and IL6 and the upregulation of CD55 may be associated with suppression of inflammation. An increase in VEGFA and BMP2 gene expression suggests that fibroblasts respond to zoledronic acid by producing a proangiogenic environment.
颌骨坏死被认为是接受双膦酸盐治疗患者的一种严重并发症。双膦酸盐的抗血管生成作用被认为与双膦酸盐相关颌骨坏死（BRONJ）的发病机制有关。本研究的目的是确定唑来膦酸对培养的人牙龈成纤维细胞的影响，以及与血管生成调节相关基因的调控情况。
从三名患者在牙冠延长手术中切除的牙龈组织中培养原代成纤维细胞。细胞分别在添加和不添加30μM唑来膦酸的条件下培养6、12和24小时，然后分别使用CellTiter - Blue和划痕伤口试验研究细胞增殖和迁移情况。使用半定量PCR阵列技术测定基因表达，该技术可分析84个已知在血管生成调节中起重要作用的通路相关基因。
唑来膦酸在暴露12和24小时时以剂量依赖方式增加牙龈成纤维细胞的增殖。人牙龈成纤维细胞划痕损伤后，用递增剂量和时间暴露于唑来膦酸（ZA）进行处理可抑制其迁移。在30μM唑来膦酸作用下，RHOB、VEGFA、CD55和BMP2的基因表达有统计学意义的增加（p≤0.05）。CCL2和IL6基因显著下调（p≤0.05）。
本研究中异戊二烯化蛋白RHOB的调节与唑来膦酸对甲羟戊酸途径的已知作用一致。CCL2和IL6的下调以及CD55的上调可能与炎症抑制有关。VEGFA和BMP2基因表达的增加表明成纤维细胞通过产生促血管生成环境对唑来膦酸作出反应。
