In vitro wound healing improvement by low-level laser therapy application in cultured gingival fibroblasts.
In vitro wound healing improvement by low-level laser therapy application in cultured gingival fibroblasts.
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DOI：
10.1155/2012/719452
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发表时间：
2012
影响因子：
2.1
通讯作者：
de Souza Costa CA
中科院分区：
文献类型：
--
作者：
Basso FG;Pansani TN;Turrioni AP;Bagnato VS;Hebling J;de Souza Costa CA
本研究的目的是利用LLLT的特定参数确定适当的能量剂量，以产生对人牙龈成纤维细胞培养的生物刺激效应。将细胞(3×104个/cm2)接种于24孔丙烯酸培养板上，普通DMEM+10%胎牛血清。在37°C与5%CO2孵育48小时后，用InGaAsP半导体激光样机(LASERTable；780±3 nm；40 mW)照射，能量剂量分别为0.5、1.5、3、5和7 J/cm~2。每24 h照射一次细胞，共照射3次。最后一次照射后2 4h，用四甲基偶氮唑盐比色法测定细胞代谢，选择最有效的两个剂量(0.5和3 J/cm2)检测细胞数(台盼蓝实验)和细胞迁移能力(伤口愈合实验和跨孔迁移实验)。数据分析采用Kruskal-Wallis和Mann-Whitney非参数检验，统计学意义为5%。0.5cm2和3cm2 J/cm2照射组细胞代谢率明显高于未照射组(P<；0.05)。两种能量剂量均能显著增加细胞数量和细胞迁移能力(P<；0.05)。这些结果表明，在所测试的条件下，LLLT在体外促进成纤维细胞的生物刺激。
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 × 104 cells/cm2) were seeded on 24-well acrylic plates using plain DMEM supplemented with 10% fetal bovine serum. After 48-hour incubation with 5% CO2 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/cm2. 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/cm2) 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/cm2 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.