Fávaro-Pípi Elaine, Ribeiro Daniel Araki, Ribeiro Juliana Uema, Bossini Paulo, Oliveira Poliani, Parizotto Nivaldo A, Tim Carla, de Araújo Heloísa Sobreiro Selistre, Renno Ana Claudia Muniz
Department of Physiotherapy, Federal University of São Carlos, Brazil.
Photomed Laser Surg. 2011 May;29(5):311-7. doi: 10.1089/pho.2010.2841. Epub 2011 Feb 9.
The aim of this study was to measure the temporal pattern of the expression of osteogenic genes after low-level laser therapy during the process of bone healing. We used quantitative real-time polymerase chain reaction (qPCR) along with histology to assess gene expression following laser irradiation on created bone defects in tibias of rats.
The animals were randomly distributed into two groups: control or laser-irradiated group. Noncritical size bone defects were surgically created at the upper third of the tibia. Laser irradiation started 24 h post-surgery and was performed for 3, 6, and 12 sessions, with an interval of 48 h. A 830 nm laser, 50 J/cm(2), 30 mW, was used. On days 7, 13, and 25 post-injury, rats were sacrificed individually by carbon dioxide asphyxia. The tibias were removed for analysis.
The histological results revealed intense new bone formation surrounded by highly vascularized connective tissue presenting slight osteogenic activity, with primary bone deposition in the group exposed to laser in the intermediary (13 days) and late stages of repair (25 days). The quantitative real-time PCR showed that laser irradiation produced an upregulation of BMP-4 at day 13 post-surgery and an upregulation of BMP4, ALP, and Runx 2 at day 25 after surgery.
Our results indicate that laser therapy improves bone repair in rats as depicted by differential histopathological and osteogenic genes expression, mainly at the late stages of recovery.
本研究旨在测量低强度激光治疗后骨愈合过程中成骨基因表达的时间模式。我们使用定量实时聚合酶链反应(qPCR)以及组织学方法来评估激光照射大鼠胫骨所造成的骨缺损后的基因表达情况。
将动物随机分为两组:对照组和激光照射组。在胫骨上三分之一处通过手术制造非临界尺寸的骨缺损。术后24小时开始激光照射,共进行3次、6次和12次照射,间隔为48小时。使用波长830nm、能量密度50J/cm²、功率30mW的激光。在受伤后第7天、13天和25天,分别通过二氧化碳窒息法处死大鼠。取出胫骨进行分析。
组织学结果显示,在激光照射组的修复中期(13天)和后期(25天),有强烈的新骨形成,周围是血管高度丰富的结缔组织,呈现轻微的成骨活性,有初级骨沉积。定量实时PCR结果显示,激光照射在术后第13天使BMP - 4表达上调,在术后第25天使BMP4、ALP和Runx 2表达上调。
我们的结果表明,如不同的组织病理学和成骨基因表达所示,激光治疗可改善大鼠的骨修复,主要是在恢复后期。