The effect of low-intensity pulsed ultrasound on wound healing using scratch assay in epithelial cells
The effect of low-intensity pulsed ultrasound on wound healing using scratch assay in epithelial cells
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DOI：
10.1016/j.jpor.2016.03.002
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发表时间：
2016-10-01
影响因子：
3.6
通讯作者：
Hosokawa, Ryuji
中科院分区：
文献类型：
--
作者：
Iwanabe, Yujiro;Masaki, Chihiro;Hosokawa, Ryuji
目的：低强度脉冲超声(LIPUS)在骨折治疗中缩短生物创面愈合所需时间，在医学领域得到广泛应用。此外，在牙科领域，LIPUS应该被有效地用于种植治疗，但大多数相关报道都是关于LIPUS对种植体周围骨形成的影响，而LIPUS对软组织愈合的影响尚不清楚。在本研究中，我们利用牙龈上皮细胞来研究LIPUS对软组织愈合的影响。方法：在培养皿中培养牙龈上皮细胞，然后LIPUS以3 MHz的频率照射15min。用血细胞计数仪计数细胞数，用显微镜测量划痕伤口闭合面积进行划痕试验。在LIPUS暴露后，收集总RNA用于微阵列分析。采用实时荧光定量聚合酶链式反应检测细胞整合素α6β4的表达水平，并收集细胞总蛋白，用免疫印迹法检测整合素α6β4的蛋白表达水平。结果：细胞计数和划痕实验结果表明，LIPUS照射促进了细胞的增殖和划痕愈合。微阵列分析表明，包括整合素在内的粘连相关基因的表达水平增加。实时定量聚合酶链式反应分析显示，LIPUS暴露可显著上调整合素α6β4的mRNA表达水平，Western blotting显示整合素α6β4表达增强。结论：与对照组相比，Lipus暴露可促进创面愈合，上调整合素α6β4的表达水平。(C)2016年日本口腔修复学会。爱思唯尔有限公司出版。保留所有权利。
Purpose: Low-intensity pulsed ultrasound (LIPUS) is widely used in medical fields because it shortens the time required for biologic wound healing in fracture treatment. Also, in dental fields, LIPUS should be effectively employed for implant treatment.However, most of the relevant reports have been published on its effects on bone formation around implants, and the effects of LIPUS on soft tissue healing remain unclear. In the present study, we examined the effects of LIPUS on soft tissue healing using gingival epithelial cells.Methods: Gingival epithelial cells were cultured on a dish, followed by LIPUS exposure at a frequency of 3 MHz for 15 min. The cells were counted with a hemocytometer, and a scratch assay was conducted by measuring the closing area of the scratch wound using a microscope. Following LIPUS exposure, total RNA was collected for microarray analysis. In addition, real-time PCR was performed to examine the mRNA expression level of integrin alpha 6 beta 4. Furthermore, total protein was collected to examine the protein expression level of integrin alpha 6 beta 4 by western blotting.Results: The cell count and scratch assay demonstrated that LIPUS exposure promoted cell proliferation and scratch-wound closure. Microarray analysis demonstrated the increased expression levels of adhesion-related genes, including integrin. Real-time PCR analysis demonstrated that LIPUS exposure significantly up-regulated the mRNA expression level of integrin alpha 6 beta 4. Western blotting showed intense staining of integrin alpha 6 beta 4.Conclusion: LIPUS exposure promotes wound closure in the scratch assay and up-regulates the expression level of integrin alpha 6 beta 4 as compared with the control. (C) 2016 Japan Prosthodontic Society. Published by Elsevier Ltd. All rights reserved.