Edelmayer Michael, Al-Habbal Diana, Pensch Manuela, Janjić Klara, Agis Hermann
1 Department of Oral Surgery, School of Dentistry, Medical University of Vienna, Vienna, Austria.
2 Department of Conservative Dentistry and Periodontology, School of Dentistry, Medical University of Vienna, Vienna, Austria.
J Biomater Appl. 2017 May;31(10):1370-1379. doi: 10.1177/0885328217702563. Epub 2017 Apr 4.
Prolyl hydroxylase inhibitors induce a proangiogenic response and are therefore proposed to optimize regenerative approaches in periodontics and oral surgery. Here the effect of the prolyl hydroxylase inhibitors dimethyloxalylglycine and deferoxamine, released from collagen barrier membranes, on osteoclastogenesis and osteoblastogenesis was evaluated. Collagen barrier membranes were loaded with dimethyloxalylglycine and deferoxamine. Release studies were performed and supernatants were taken after 1, 3, 6, 24, and 48 h. The effect of these supernatants on osteoblast- and osteoclast-precursor cells was evaluated. Furthermore, dose response studies for dimethyloxalylglycine and deferoxamine were performed. Osteoclastogenesis was evaluated with RAW 264.7 cells based on the number of multinuclear tartrate-resistant acid phosphatase positive cells. Osteoblastogenesis was evaluated with MC3T3-E1 cells based on alkaline phosphatase. Metabolic activity and cell proliferation were assessed based on MTT and BrdU assays. Vascular endothelial growth factor production was evaluated using an immunoassay. We found that supernatants taken in the first hour from collagen barrier membranes loaded with dimethyloxalylglycine or deferoxamine reduced osteoclastogenesis. Osteoblastogenesis was not reduced significantly. Cell proliferation and metabolic activity of RAW 264.7 and MC3T3-E1 cells were inhibited by supernatants of collagen barrier membranes loaded with deferoxamine but not dimethyloxalylglycine. In RAW 264.7 cell culture, vascular endothelial growth factor production was increased only by supernatants of collagen barrier membranes loaded with dimethyloxalylglycine, but not deferoxamine. In MC3T3-E1 cell culture, supernatants of collagen barrier membranes loaded with dimethyloxalylglycine and deferoxamine both increased vascular endothelial growth factor production. Direct measurements showed that the majority of dimethyloxalylglycine and deferoxamine is released in the first hours. Dose-response studies supported the divergent effects of deferoxamine and dimethyloxalylglycine in RAW 264.7 and MC3T3-E1 cultures. Our findings show diverse effects of dimethyloxalylglycine- and deferoxamine-loaded collagen barrier membranes during osteoclastogenesis and osteoblastogenesis. Preclinical studies will reveal if the increase in vascular endothelial growth factor together with the inhibitory effect on osteoclasts can stimulate oral tissue regeneration.
脯氨酰羟化酶抑制剂可诱导促血管生成反应，因此被提议用于优化牙周病学和口腔外科的再生方法。在此，评估了从胶原屏障膜释放的脯氨酰羟化酶抑制剂二甲基草酰甘氨酸和去铁胺对破骨细胞生成和成骨细胞生成的影响。胶原屏障膜加载二甲基草酰甘氨酸和去铁胺。进行释放研究，并在1、3、6、24和48小时后收集上清液。评估这些上清液对成骨细胞和破骨细胞前体细胞的影响。此外，进行了二甲基草酰甘氨酸和去铁胺的剂量反应研究。基于抗酒石酸酸性磷酸酶阳性多核细胞的数量，用RAW 264.7细胞评估破骨细胞生成。基于碱性磷酸酶，用MC3T3-E1细胞评估成骨细胞生成。基于MTT和BrdU测定评估代谢活性和细胞增殖。使用免疫测定评估血管内皮生长因子的产生。我们发现，在加载二甲基草酰甘氨酸或去铁胺的胶原屏障膜的第一小时收集的上清液可减少破骨细胞生成。成骨细胞生成没有显著减少。加载去铁胺而非二甲基草酰甘氨酸的胶原屏障膜的上清液可抑制RAW 264.7和MC3T3-E1细胞的细胞增殖和代谢活性。在RAW 264.7细胞培养中，仅加载二甲基草酰甘氨酸而非去铁胺的胶原屏障膜的上清液可增加血管内皮生长因子的产生。在MC3T3-E1细胞培养中，加载二甲基草酰甘氨酸和去铁胺的胶原屏障膜的上清液均可增加血管内皮生长因子的产生。直接测量表明，大部分二甲基草酰甘氨酸和去铁胺在最初几小时内释放。剂量反应研究支持了去铁胺和二甲基草酰甘氨酸在RAW 264.7和MC3T3-E1培养中的不同作用。我们的研究结果表明，加载二甲基草酰甘氨酸和去铁胺的胶原屏障膜在破骨细胞生成和成骨细胞生成过程中具有不同的作用。临床前研究将揭示血管内皮生长因子的增加以及对破骨细胞的抑制作用是否能刺激口腔组织再生。
