目的:建立犬声带瘢痕模型,分析损伤后不同时间段内声带愈合的形态学、组织病理学及细胞外基质(ECM)相关影响因子的变化。方法:5只实验犬随机分为对照组(1只)和实验组(4只),对照组不做特殊处理,实验组行支撑喉镜下CO_2激光双侧声带烧灼术。各只实验犬分别在5个不同时间(术前和术后6h、3周、8周和12周)于支撑喉镜下观察声带损伤的大体形态学变化后处死,取材。采用苏木精-伊红染色、免疫荧光等方法观察声带各层组织病理学变化和超微形态学变化。观测声带组织内赖氨酰氧化酶(LOX)、热休克蛋白70(HSP70)和透明质酸(HA)的累积光密度值(IOD)的变化规律。结果:1喉内镜示,损伤后3周声带创面轻度充血水肿及炎性渗出;术后8周创面充血水肿消失,局部挛缩凹陷,无粘连和肉芽形成;术后12周双侧声带表面光滑,局部挛缩,声带瘢痕形成,声嘶明显。2苏木精-伊红染色示,术后6h声带创面大量炎性细胞浸润、红细胞渗出,细胞充血水肿;术后3周声带创面成纤维细胞增生,新生血管形成,大量纤维组织无序排列;术后8周声带创面各层纤维组织增生紊乱,血管增粗;术后12周时,声带创面固有层增厚,大量胶原纤维增多,排列紊乱,呈团状或束状,纤维间较多不规则间隙;血管和腺体少见或消失。3免疫荧光示LOX、HSP70、HA均定位于细胞质和细胞核,术后不同时间段表达的阳性程度不同,大多在炎性细胞、血管内皮细胞和腺体内表达较强烈。4IOD值比较,在不同时间段LOX的IOD值之间均有差异(P<0.05);HSP70在术前与术后12周的IOD值无明显异常,余各组间差异有统计学意义(P<0.01);HA在术后3周与12周间无异常,余各组间差异有统计学意义(P<0.05)。5LOX在术后6h表达减弱,术后3~12周呈上升趋势;HSP70在术后6h升至峰值,术后3~12周呈下降趋势;HA在术后6h表达减弱,术后8周升至峰值,术后8~12周下降。6透射电镜显示,声带术后3~8周固有层成纤维细胞极度活跃状态,细胞肿胀,细胞器丰富;术后12周时,声带固有层内仍功能活跃,纤维细胞呈网状排列紊乱,纤维间隙较大。结论:支撑喉镜下CO_2激光声带烧灼术建立犬声带瘢痕模型可靠,损伤后3个月声带瘢痕基本形成。LOX、HSP70、HA在声带瘢痕形成的早中晚期发挥着不同的作用,可以作为检测声带瘢痕形成较敏感的指标。 OBJECTIVE: To establish a model of vocal cord scars in dogs and analyze the changes of vocal cord healing morphological, histopathological and extracellular matrix (ECM) -related factors in different time periods after injury. Methods: Five experimental dogs were randomly divided into control group (n = 1) and experimental group (n = 4). The control group received no special treatment. The experimental group received bilateral vocal cords cauterization of CO_2 laser under the support of laryngoscope. The dogs were sacrificed at 5 different times (preoperative and postoperative 6h, 3weeks, 8weeks and 12weeks) under the support of laryngoscope to observe the general morphological changes of vocal cord injury. Hematoxylin-eosin staining, immunofluorescence and other methods were used to observe the changes of histopathological and ultrastructural changes of vocal folds. The changes of cumulative optical density (IOD) of lysyl oxidase (LOX), heat shock protein 70 (HSP70) and hyaluronic acid (HA) in the vocal cords were observed. Results: 1 The laryngeal endoscope showed slight hyperemia, edema and inflammatory exudation of vocal cord wound 3 weeks after injury; the congestive and edema of wound disappeared 8 weeks after operation, the local contracture depression, no adhesions and granulation formation; bilateral vocal folds Smooth surface, local contracture, vocal cord scar formation, obvious hoarseness. 2 hematoxylin-eosin staining, a large number of inflammatory cell infiltration, erythrocyte exudation, cell congestion and edema at 6 h after operation; fibroblast hyperplasia, neovascularization and a large number of fibrous tissue disordered ; At 8 weeks after operation, the proliferation of fibrous tissue in each layer of vocal cord was disordered and the blood vessels became thick; at 12 weeks after operation, the lamina propria was thickened, a large number of collagen fibers were increased, disordered, clumped or bundled, Irregular gap; blood vessels and glands rare or disappear. Immunofluorescence showed that LOX, HSP70 and HA were localized in the cytoplasm and nucleus. The expression of LOX, HSP70 and HA were different at different time points after surgery. Most of them expressed more strongly in inflammatory cells, vascular endothelial cells and glands. (P <0.05). The IOD values ​​of HSP70 in preoperation and postoperative 12 weeks were not significantly different (P> 0.05). There were significant differences in IOD values ​​among different groups <0.01). There was no difference between the 3 weeks and 12 weeks of HA in the HA group, with significant difference (P <0.05). The expression of 5LOX was weakened at 6h after operation and increased from 3 to 12 weeks after operation. HSP70 peaked at 6h after operation and decreased from 3 to 12 after operation. The expression of HA decreased 6h after operation, To peak, after 8 to 12 weeks decreased. Transmission electron microscopy showed that the lamina propria fibroblasts became extremely active at 3 to 8 weeks after vocal cord surgery, with swollen cells and abundant organelles. At 12 weeks after operation, the lamina propria was still active in the lamina propria, Gap larger. Conclusion: The model of vocal cords scar formation with CO_2 laser vocal cord cauterization under the support of laryngoscope is reliable. The vocal cord scar formation is basically formed 3 months after injury. LOX, HSP70 and HA play different roles in the early and middle stages of vocal cord scar formation, which can be used as a sensitive index to detect vocal cord scar formation.