激光辅助鼓膜穿孔再生的细胞与组织机制

文献信息

中文摘要

背景： 慢性鼓膜（TM）穿孔由于自发愈合有限以及手术干预的缺点，仍然是一个重要的临床问题。本研究旨在评估低强度红外激光照射对再生细胞机制的影响，并评估其在TM修复中的治疗潜力。方法：将从龙猫组织分离的原代成纤维细胞培养物暴露于波长为1560 nm和1450 nm的激光辐射。使用MTT测定、划痕试验、Ki-67免疫染色和II型胶原ELISA分析细胞反应。此外，使用龙猫慢性TM穿孔的体内模型来验证生理条件下的再生效应。结果：在选定的照射条件下，激光照射与划痕闭合加速和细胞外基质产生增加相关，而代谢和增殖指数随时间下降，最可能反映培养汇合和成熟而非细胞毒性。在体内，在胶原海绵辅助修复的背景下，所有激光治疗的耳朵均观察到鼓膜穿孔完全闭合，而仅支架对照穿孔仍未闭合。结论：这些发现支持低强度红外激光照射作为支架辅助鼓膜修复的微创辅助手段的潜力。然而，所提出的内源性祖细胞群参与应被视为一个需要进一步组织学和基于标志物验证的假设。

英文摘要

Chronic tympanic membrane (TM) perforations remain a significant clinical problem due to limited spontaneous healing and the drawbacks of surgical interventions. The present study aimed to evaluate the effects of low-intensity infrared laser irradiation on cellular mechanisms of regeneration and to assess its therapeutic potential in TM repair. Primary fibroblast cultures isolated from chinchilla tissue were exposed to laser radiation at wavelengths 1560 nm and 1450 nm. Cellular responses were analyzed using MTT assay, scratch test, Ki-67 immunostaining, and ELISA for collagen type II. In addition, an in vivo model of chronic TM perforation in chinchillas was used to validate the regenerative effects under physiological conditions. Laser irradiation was associated with accelerated scratch closure and increased extracellular matrix production under selected irradiation conditions, whereas metabolic and proliferative indices decreased over time, most likely reflecting culture confluence and maturation rather than cytotoxicity. In vivo, complete closure of tympanic membrane perforations was observed in all laser-treated ears in the context of collagen sponge-assisted repair, whereas scaffold-only control perforations remained unclosed. These findings support the potential of low-intensity infrared laser irradiation as a minimally invasive adjunct to scaffold-assisted tympanic membrane repair. However, the proposed involvement of endogenous progenitor cell populations should be regarded as a hypothesis requiring further histological and marker-based validation.