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低强度激光疗法应用于培养的牙龈成纤维细胞对体外伤口愈合的改善作用

In vitro wound healing improvement by low-level laser therapy application in cultured gingival fibroblasts.

作者信息

Basso Fernanda G, Pansani Taisa N, Turrioni Ana Paula S, Bagnato Vanderlei S, Hebling Josimeri, de Souza Costa Carlos A

机构信息

Faculdade de Odontologia de Piracicaba, Universidade Estadual de Campinas (UNICAMP), 13414-903 Piracicaba, SP, Brazil.

出版信息

Int J Dent. 2012;2012:719452. doi: 10.1155/2012/719452. Epub 2012 Jul 15.

Abstract

The aim of this study was to determine adequate energy doses using specific parameters of LLLT to produce biostimulatory effects on human gingival fibroblast culture. Cells (3 × 10(4) cells/cm(2)) were seeded on 24-well acrylic plates using plain DMEM supplemented with 10% fetal bovine serum. After 48-hour incubation with 5% CO(2) at 37°C, cells were irradiated with a InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm; 40 mW) with energy doses of 0.5, 1.5, 3, 5, and 7 J/cm(2). Cells were irradiated every 24 h totalizing 3 applications. Twenty-four hours after the last irradiation, cell metabolism was evaluated by the MTT assay and the two most effective doses (0.5 and 3 J/cm(2)) were selected to evaluate the cell number (trypan blue assay) and the cell migration capacity (wound healing assay; transwell migration assay). Data were analyzed by the Kruskal-Wallis and Mann-Whitney nonparametric tests with statistical significance of 5%. Irradiation of the fibroblasts with 0.5 and 3 J/cm(2) resulted in significant increase in cell metabolism compared with the nonrradiated group (P < 0.05). Both energy doses promoted significant increase in the cell number as well as in cell migration (P < 0.05). These results demonstrate that, under the tested conditions, LLLT promoted biostimulation of fibroblasts in vitro.

摘要

本研究的目的是利用低强度激光疗法(LLLT)的特定参数确定适当的能量剂量,以对人牙龈成纤维细胞培养产生生物刺激作用。使用补充有10%胎牛血清的普通杜氏改良 Eagle 培养基(DMEM),将细胞(3×10⁴个细胞/cm²)接种到24孔丙烯酸板上。在37°C、5%二氧化碳条件下孵育48小时后,用InGaAsP二极管激光原型(LASERTable;780±3nm;40mW)以0.5、1.5、3、5和7J/cm²的能量剂量照射细胞。每24小时照射一次,共照射3次。最后一次照射后24小时,通过MTT法评估细胞代谢,并选择两个最有效的剂量(0.5和3J/cm²)来评估细胞数量(台盼蓝法)和细胞迁移能力(伤口愈合试验;Transwell迁移试验)。数据采用Kruskal-Wallis和Mann-Whitney非参数检验进行分析,统计学显著性为5%。与未照射组相比,用0.5和3J/cm²照射成纤维细胞导致细胞代谢显著增加(P<0.05)。这两个能量剂量均促进了细胞数量和细胞迁移的显著增加(P<0.05)。这些结果表明,在测试条件下,低强度激光疗法在体外促进了成纤维细胞的生物刺激。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/724f/3403309/96c065e8d632/IJD2012-719452.001.jpg

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