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目的:研究不同时间,甲苯胺蓝(Toluidine Blue O,TBO)在大鼠炎症性口腔颊粘膜渗透的浓度变化,及甲苯胺蓝与炎症细胞分布之间的关系。方法:实验选取wistar大鼠32只,炎症组大鼠20只,建立以金葡菌为优势菌的感染炎症创口模型。将浓度为1mg/mL甲苯胺蓝溶液置于大鼠感染炎症创口组织上5、10分钟后处死,正常粘膜组大鼠8只,于甲苯胺蓝在正常颊粘膜渗透5、10、20、40分钟后处死,避光条件下取组织块进行冰冻切片,即刻荧光显微镜下观察荧光分布;冰冻切片进行HE染色,观察炎症细胞分布。采用Image Pro-plus 6.0软件检测荧光分布的光密度、分布面积以及炎症分布面积。结果:1创口周围正常粘膜及正常完整粘膜组的甲苯胺蓝均停留在角化层,未穿透上皮层,和时间无相关关系;2炎症5分钟组平均荧光分布可达到炎症细胞分布面积的89%,炎症10分钟组可达108%;炎症组在创口表面及深部,荧光光密度均无显著差异。结论:1甲苯胺蓝可有效分布于感染的炎症组织,但不能穿透正常组织,完整上皮可保护正常组织免受光动力的杀伤。2浓度为1 mg/mL的甲苯胺蓝溶液渗透时间为10分钟时,创口中甲苯胺蓝的分布与炎症细胞的分布基本一致,甲苯胺蓝浓度梯度无显著变化。提示甲苯胺蓝作为光敏剂在针对口腔创口感染的抗菌光动力疗法中可有效、安全的发挥作用。
OBJECTIVE: To study the changes of Toluidine Blue O (TBO) concentration in inflamed oral buccal mucosa and the relationship between toluidine blue and the distribution of inflammatory cells at different time points. Methods: Twenty-two wistar rats and 20 inflammatory rats were selected to establish an inflammatory wound model with Staphylococcus aureus as dominant bacterium. Toluidine blue solution at a concentration of 1 mg / mL was placed on the wound tissue of infected rats for 5 and 10 minutes, then sacrificed. Eight rats in the normal mucosa group were infused with toluidine blue in normal buccal mucosa 5, 10, 20, 40 Minutes after sacrifice, under dark conditions, take tissue slices frozen sections, immediately observed under fluorescent microscope fluorescence distribution; frozen sections were HE staining to observe the distribution of inflammatory cells. Image Pro-plus 6.0 software was used to detect the optical density, distribution area and distribution area of the fluorescence distribution. Results: 1 Toluidine blue in the normal mucosa and normal intact mucosa of the wound all stayed in the keratinized layer and did not penetrate the epithelial layer, and had no correlation with the time; 2 The average fluorescence distribution in 5 minutes of inflammation could reach the distribution area of inflammatory cells 89%, inflammation 10 minutes group up to 108%; inflammation in the wound surface and deep, no significant difference in fluorescence optical density. Conclusion: Toluidine blue can be effectively distributed in the infected inflammatory tissue, but can not penetrate the normal tissue, the intact epithelium can protect the normal tissue from photodynamic killing. 2 toluidine blue solution concentration of 1 mg / mL infiltration time of 10 minutes, the wound toluidine blue distribution and inflammatory cell distribution is basically the same, toluidine blue concentration gradient no significant change. Toluidine blue as a photosensitizer prompted anti-bacterial photodynamic therapy for oral wound infection can be effective and safe to play a role.