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660nm 和 980nm 半导体激光对牙周膜间充质干细胞分化的影响。

Effect of photobiomodulation therapy with 660 and 980 nm diode lasers on differentiation of periodontal ligament mesenchymal stem cells.

机构信息

Department of Periodontics, Faculty of Dentistry, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.

Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Sci Rep. 2024 Sep 4;14(1):20587. doi: 10.1038/s41598-024-71386-3.

Abstract

This study aimed to compare the effects of photobiomodulation therapy (PBMT) with 660 and 980 nm diode lasers on differentiation of periodontal ligament mesenchymal stem cells (PDLMSCs). In this in vitro, experimental study, PDLMSCs were obtained from the Iranian Genetic Bank and cultured in osteogenic medium. They were then subjected to irradiation of 660 and 980 nm diode lasers, and their viability was assessed after one, two, and three irradiation cycles using the methyl thiazolyl tetrazolium (MTT) assay. The cells also underwent DAPI staining, cell apoptosis assay by using the Annexin V/PI, Alizarin Red staining, and real-time polymerase chain reaction (PCR) for assessment of the expression of osteogenic genes. Data were analyzed by two-way ANOVA. The two laser groups had no significant difference in cell apoptosis according to the results of DAPI staining. Both laser groups showed higher cell viability in the MTT assay at 4 and 6 days compared with the control group. Annexin V/PI results showed higher cell viability in both laser groups at 4 days compared with the control group. Rate of early and late apoptosis was lower in both laser groups than the control group at 4 days. Necrosis had a lower frequency in 980 nm laser group than the control group on day 6. Alizarin Red staining showed higher cell differentiation in both laser groups after 3 irradiation cycles than the control group. The highest expression of osteopontin (OPN), osteocalcin (OCN), and Runt-related transcription factor 2 (RUNX2) was noted in 660 nm laser group with 3 irradiation cycles at 14 days, compared with the control group. PBMT with 660 and 980 nm diode lasers decreased apoptosis and significantly increased PDLMSC differentiation after 3 irradiation cycles.

摘要

本研究旨在比较 660nm 和 980nm 半导体激光对牙周膜间充质干细胞(PDLMSCs)分化的影响。在这项体外实验研究中,PDLMSCs 从伊朗遗传银行获得并在成骨培养基中培养。然后,用 660nm 和 980nm 半导体激光照射细胞,并在 1、2 和 3 个照射周期后用噻唑蓝(MTT)法评估细胞活力。还对细胞进行 DAPI 染色、用 Annexin V/PI 进行细胞凋亡检测、茜素红染色和实时聚合酶链反应(PCR)评估成骨基因的表达。采用双因素方差分析进行数据分析。根据 DAPI 染色结果,两组激光细胞凋亡无显著差异。与对照组相比,两组激光在 MTT 检测中在 4 天和 6 天时的细胞活力更高。 Annexin V/PI 结果表明,与对照组相比,两组激光在 4 天时的细胞活力更高。与对照组相比,两组激光在 4 天时的早期和晚期凋亡率较低。与对照组相比,980nm 激光组在第 6 天的坏死频率较低。茜素红染色显示,与对照组相比,两组激光在 3 个照射周期后细胞分化更高。与对照组相比,在 14 天时,660nm 激光组在 3 个照射周期后骨桥蛋白(OPN)、骨钙素(OCN)和 runt 相关转录因子 2(RUNX2)的表达最高。660nm 和 980nm 半导体激光的 PBMT 可减少细胞凋亡,并在 3 个照射周期后显著增加 PDLMSC 分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb99/11375153/a9dbd6fd6eb8/41598_2024_71386_Fig1_HTML.jpg

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