P M Mandrillo, G Fischetto, P Odorisio, F Cura, A Avantaggiato, F Carinci
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引用次数: 27
Abstract
Purpose: The light-emitting diodes (LEDs) have been applied in oral surgery for tissue stimulation and wound healing. Several Authors have highlighted that fibroblasts subjected to phototherapy have an increased viability, proliferation, biomodulation of inflammatory cytokines and genes expression. It remains to be determined which are the best irradiation parameters (energy, wavelength, power) for each type of cell in order to obtain the best bio-stimulation. The aim of this study was to investigate the effects of LED irradiation on primary human gingival fibroblast cells (HGF) on DSP, ELN, HAS1, ELANE, HYAL1, RPL13 genes activation using Real Time PCR. These genes activation is directly connected with elastin protein production and HGF functionality.
Materials and methods: Human gingival tissue biopsies were obtained from three healthy patients during extraction of teeth. The gingival pieces were fragmented with a scalpel and transferred in culture dishes for allow the cells growth. Human gingival fibroblasts at the second passage were seeded on multiple 6-well plates and were stimulated with three different light-emitting diodes (LEDs) fixture. After irradiation, the cells were trypsinized, harvested and lysed for RNA extraction. Genes expression was quantified using Real Time PCR.
Results: We didn't found significant differences in genes activation of HGF of the three different LEDs. The LED irradiation seems to be directly correlated with the elastin and hyaluronoglucosaminidase 1 genes activation that are directly connected with proteins production and HGF functionality.
Conclusions: HGF show an increased deposition of elastin as well as enhanced expression of collagen type I, which is the main protein related to the synthesis and of the collagen-rich matrix.
目的:发光二极管(led)在口腔外科中用于组织刺激和伤口愈合。一些作者强调,受光疗的成纤维细胞具有更高的活力、增殖、炎症细胞因子的生物调节和基因表达。为了获得最佳的生物刺激,每种细胞的最佳辐照参数(能量、波长、功率)仍有待确定。本研究采用Real Time PCR技术研究LED辐照对人原代牙龈成纤维细胞(HGF)中DSP、ELN、HAS1、ELANE、HYAL1、RPL13等基因激活的影响。这些基因的激活与弹性蛋白的产生和HGF的功能直接相关。材料与方法:对3例健康患者拔牙时的牙龈组织进行活检。用手术刀将牙龈碎片破碎,并将其转移到培养皿中以使细胞生长。将第二代人牙龈成纤维细胞播种在多个6孔板上,并使用三种不同的发光二极管(led)灯具进行刺激。照射后,细胞胰蛋白酶化,收获和裂解RNA提取。采用Real Time PCR定量检测基因表达。结果:我们发现三种不同的led在HGF的基因活化方面没有显著差异。LED照射似乎与弹性蛋白和透明质糖氨基酶1基因激活直接相关,而弹性蛋白和透明质糖氨基酶1基因激活与蛋白质产生和HGF功能直接相关。结论:HGF显示弹性蛋白沉积增加,I型胶原表达增强,而I型胶原是富胶原基质合成的主要蛋白。