Kargarpour Zahra, Nasirzade Jila, Di Summa Francesca, Panahipour Layla, Miron Richard J, Gruber Reinhard
Department of Oral Biology, Medical University of Vienna, 1090 Vienna, Austria.
Department of Periodontology, School of Dental Medicine, University of Bern, 3012 Bern, Switzerland.
Antioxidants (Basel). 2020 Jun 26;9(6):560. doi: 10.3390/antiox9060560.
Hydrogen peroxide is a damage signal at sites of chronic inflammation. The question arises whether platelet-rich fibrin (PRF), platelet-poor plasma (PPP), and the buffy coat can neutralize hydrogen peroxide toxicity and thereby counteract local oxidative stress. In the present study, gingival fibroblasts cells were exposed to hydrogen peroxide with and without lysates obtained from PRF membranes, PPP, heated PPP (75 °C for 10 min), and the buffy coat. Cell viability was examined by trypan blue staining, live-dead staining, and formazan crystal formation. Cell apoptosis was assessed by cleaved caspase-3 Western blot analysis. Reverse transcription-quantitative polymerase chain reaction (RT-PCR) was utilized to determine the impact of PRF lysates on the expression of catalase in fibroblasts. It was reported that lysates from PRF, PPP, and the buffy coat-but not heated PPP-abolished the hydrogen peroxide-induced toxicity in gingival fibroblasts. Necrosis was confirmed by a loss of membrane integrity and apoptosis was ruled out by the lack of cleavage of caspase-3. Aminotriazole, an inhibitor of catalase, reduced the cytoprotective activity of PRF lysates yet blocking of glutathione peroxidase by mercaptosuccinate did not show the same effect. PRF lysates had no impact on the expression of catalase in gingival fibroblasts. These findings suggest that PRF, PPP, and the buffy coat can neutralize hydrogen peroxide through the release of heat-sensitive catalase.
过氧化氢是慢性炎症部位的一种损伤信号。由此产生的问题是，富血小板纤维蛋白（PRF）、贫血小板血浆（PPP）和血沉棕黄层是否能够中和过氧化氢毒性，从而对抗局部氧化应激。在本研究中，将牙龈成纤维细胞暴露于过氧化氢中，同时添加或不添加从PRF膜、PPP、加热的PPP（75℃ 10分钟）和血沉棕黄层获得的裂解物。通过台盼蓝染色、活死染色和甲臜晶体形成来检测细胞活力。通过裂解的半胱天冬酶-3蛋白质印迹分析评估细胞凋亡。利用逆转录定量聚合酶链反应（RT-PCR）来确定PRF裂解物对成纤维细胞中过氧化氢酶表达的影响。据报道，PRF、PPP和血沉棕黄层的裂解物——而非加热的PPP——消除了过氧化氢对牙龈成纤维细胞的毒性。通过膜完整性丧失证实了坏死，并且由于半胱天冬酶-3未裂解而排除了凋亡。过氧化氢酶抑制剂氨基三唑降低了PRF裂解物的细胞保护活性，但巯基琥珀酸对谷胱甘肽过氧化物酶的阻断未显示相同效果。PRF裂解物对牙龈成纤维细胞中过氧化氢酶的表达没有影响。这些发现表明，PRF、PPP和血沉棕黄层可以通过释放热敏感的过氧化氢酶来中和过氧化氢。
