Department of Physical Therapy, Federal University of Rio Grande do Norte, Natal, Brazil.
Department of Health Sciences, Federal University of the Semiarid Region, Mossoró, Brazil.
PLoS One. 2019 Feb 4;14(2):e0211643. doi: 10.1371/journal.pone.0211643. eCollection 2019.
The aim of this study was to compare the treatment effects of laser photobiomodulation (LPBM) therapy and aerobic exercise on the biomechanical properties, tissue morphology and the expression of tendon matrix molecules during early remodeling of Achilles tendon (AT) injury in diabetic rats. Animals were randomly assigned to five groups: injured non diabetic (I, n = 15), injured diabetic (ID, n = 15), injured diabetic plus LPBM (IDL, n = 16), injured diabetic plus aerobic exercise (IDE, n = 16) and injured diabetic plus aerobic exercise and LPBM (IDEAL, n = 17). Type 1 diabetes was induced via a single intravenous injection of Streptozotocin at a dose of 40 mg/kg. A partial tenotomy was performed in the right AT. LPBM was performed with an indium-gallium-aluminum-phosphide 660 nm 10 mW laser device (spot size 0.04 cm2, power density 250 mW/cm2, irradiation duration 16 s, energy 0.16 J, energy density 4 J/cm2) on alternate days for a total of 9 sessions over 3 weeks (total energy 1.44 J), using a stationary contact technique to a single point over the dorsal aspect of the AT. Moderate aerobic exercise was performed on a motorized treadmill (velocity 9 m/min for 60 minutes). At 3 weeks post-injury, biomechanical analyzes as well as assessment of fibroblast number and orientation were performed. Collagen 1 (Col1) and 3 (Col3) and matrix metalloproteinases (MMPs) -3 and 13 protein distributions were studied by immunohistochemistry; while Col1 and Col3 and MMP-2 and 9 gene expression were assessed by quantitative RT-PCR (qRT-PCR). IDEAL exhibited significant increases in several biomechanical parameters in comparison to the other groups. Moreover, IDEAL presented stronger Col1 immunoreactivity when compared to ID, and weaker Col3 immunoreactivity than IDE. Both IDL and IDEAL demonstrated weaker expression of MMP-3 in comparison to I, while IDL presented no expression of MMP-13 when compared to ID. ID, IDL and IDE showed an increased number of fibroblasts in comparison to I, while IDEAL decreased the number of these cells in comparison to ID and IDE. IDL and IDEAL groups exhibited decreased angular dispersion among the fibroblasts when compared to I. The gene expression results showed that IDE demonstrated a downregulation in Col1 mRNA expression in comparison to I and ID. IDEAL demonstrated upregulation of Col1 mRNA expression when compared to IDL or IDE alone and increased MMP-2 expression when compared to IDL and IDE. MMP-9 expression was upregulated in IDEAL when compared to I, IDL and IDE. Our results suggest a beneficial interaction of combining both treatment strategies i.e., aerobic exercise and LPBM, on the biomechanical properties, tissue morphology and the expression of matrix molecules in diabetic tendons.
本研究旨在比较激光光生物调节(LPBM)治疗与有氧运动对糖尿病大鼠跟腱(AT)损伤早期重塑过程中生物力学特性、组织形态和腱基质分子表达的治疗效果。动物随机分为五组:非糖尿病损伤组(I,n=15)、糖尿病损伤组(ID,n=15)、糖尿病损伤加 LPBM 组(IDL,n=16)、糖尿病损伤加有氧运动组(IDE,n=16)和糖尿病损伤加有氧运动和 LPBM 组(IDEAL,n=17)。1 型糖尿病通过单次静脉注射链脲佐菌素(Streptozotocin)诱导,剂量为 40mg/kg。右侧 AT 行部分腱切开术。LPBM 采用铟镓铝磷 660nm 10mW 激光设备(光斑尺寸 0.04cm2,功率密度 250mW/cm2,辐照时间 16s,能量 0.16J,能量密度 4J/cm2)进行,每两天进行一次,共 9 次,持续 3 周(总能量 1.44J),采用固定接触技术在 AT 背侧的单个点进行照射。采用电动跑步机进行适度有氧运动(速度 9m/min,持续 60min)。损伤后 3 周,进行生物力学分析以及成纤维细胞数量和方向评估。通过免疫组织化学研究胶原 1(Col1)和 3(Col3)以及基质金属蛋白酶(MMPs)-3 和 13 蛋白分布;通过定量 RT-PCR(qRT-PCR)评估 Col1 和 Col3 以及 MMP-2 和 9 基因表达。与其他组相比,IDEAL 组的多个生物力学参数显著增加。此外,与 ID 相比,IDEAL 组 Col1 免疫反应性更强,而 Col3 免疫反应性比 IDE 组弱。与 I 相比,IDL 和 IDEAL 组 MMP-3 的表达均减弱,而与 ID 相比,IDL 组 MMP-13 无表达。ID、IDL 和 IDE 组的成纤维细胞数量与 I 组相比均增加,而 IDEAL 组的成纤维细胞数量与 ID 和 IDE 组相比减少。与 I 组相比,IDL 和 IDEAL 组的成纤维细胞角分布减少。基因表达结果表明,与 I 和 ID 相比,IDE 组 Col1mRNA 表达下调。与单独的 IDL 或 IDE 相比,IDEAL 组 Col1mRNA 表达上调,与 IDL 和 IDE 相比,MMP-2 表达上调。与 I、IDL 和 IDE 相比,IDEAL 组 MMP-9 表达上调。我们的结果表明,有氧运动和 LPBM 联合应用这两种治疗策略对糖尿病肌腱的生物力学特性、组织形态和基质分子表达具有有益的相互作用。