应用激光, 2018, 38 (1): 154, 网络出版: 2018-05-03  

HMME-PDT对大鼠口腔混合菌生物膜抑制作用的实验研究

The Experimental Study of the Effect of HMME-PDT Inhibition the Cariogenic Bacteria in Dental Plaque Biofilm of Rats
作者单位
1 承德医学院附属医院口腔科, 河北 承德 067000
2 承德医学院, 河北 承德 067300
摘要
目的: 探讨不同光敏剂浓度和不同光照时间的HMME-PDT对大鼠口腔混合菌菌斑生物膜的抑制效果, 确定最佳光敏剂参数和时间参数, 并在此基础上探讨HMME-PDT在抑制混合菌菌斑生物膜的优势, 为PDT应用于龋病的预防提供理论与实验依据。方法: 以 Wistar 大鼠构建混合菌致龋模型。将其随机分组: A组为生理盐水阴性处理组, B组为氟化钠阳性对照组, 其余组别根据光敏剂浓度的不同, 分为C1组20 mg/L, D1组30 mg/L, E1组40 mg/L, F1组50 mg/L, G1组60 mg/L, 避光孵育5 min后, 波长为532 nm的半导体激光器照射90 s, 光斑半径为0.5 cm, 功率密度为0.14 W/cm2。根据照射时间的不同, 分为C2组30 s, D2组60 s, E2组90 s, F2组120 s, G2组150 s, 光敏剂浓度40 mg/L避光孵育5 min后用波长为532 nm的半导体激光器照射, 光斑半径为0.5 cm, 功率密度为0.14 W/cm2照射。实验处理后取样, 通过平板菌落计数法观察各组对大鼠口腔混合菌生物膜的抑制作用, 原子吸收分光光度计测定光动力疗法作用后大鼠口腔混合菌生物膜的钙离子溶出量, 评价HMME-PDT对大鼠口腔混合菌菌斑生物膜的抑制作用。结果: 1.不同光敏剂浓度下HMME-PDT对大鼠口腔混合菌菌斑生物膜抑菌效果的影响。平板菌落计数结果显示: 不同光敏剂浓度的各组抑菌率随光敏剂浓度的增加而增高。与阴性对照组及阳性对照组相比E1组有明显差异。2.不同光照时间下HMME-PDT对大鼠口腔混合菌菌斑生物膜抑菌效果的影响。平板菌落计数结果显示: 不同光照时间的各组抑菌率随时间的增加而增高。与阴性对照组及阳性对照组相比E2组有明显差异。3.原子吸收分光光度计测定钙离子溶出量结果显示, 在48小时内, 各组钙离子溶出量(μg/ml)随时间的延长而增加。与阴性对照组相比PDT组各时间点的钙离子溶出量均显著减小(P<0.05)。与阳性对照组相比PDT各组均能使钙离子溶出量减小(P<0.05), 其中E1组与E2组改变最明显。结论: HMME-PDT对大鼠口腔混合菌生物膜有明显抑制作用, 实验证实HMME-PDT最佳光敏剂浓度为40 mg/L, 最佳光照时间为90 s。光动力疗法能有效抑制大鼠口腔混合菌菌斑生物膜中的致龋菌, 可减少早期龋病的钙离子溶出量, 不但阻止了釉质继续脱矿, 而且促进了釉质的再矿化。PDT在龋病的预防中具有广阔的临床应用前景。
Abstract
Object: To Study the inhibition effective of HMME-PDT in different photosensitizer concentration and different lighting time on the cariogenic bacteria in dental plaque biofilms of rats, and determine the optimal photosensitizer parameters and time parameters. Besides we explored the advantages of HMME-PDT in inhibiting the biofilm of mixed bacterial plaque. We provided theoretical and experimental basis for the application of PDT for dental caries prevention. Method: The caries model was constructed with Wistar rats. We grouped the rats in accordance with the principle of randomization as follows: Group A: a saline-negative treatment group, Group B: a positive control group for fluoride. The other groups were based on the concentration of the photosensitizer as group C1 20 mg/L, group D1 30 mg/L, group E1 40 mg/L, group F1 50 mg/L, group G1 60 mg/L. Treated group C1-G1 with incubation for 5 minutes in dark place and exposed 90 s by a semiconductor laser with a wavelength of 532 nm. The spot radius was 0.5 cm and the power density was 0.14 W/cm2. The rest groups were based on the exposure time: group C2 30 s, group D2 60 s, group E2 90 s, group F2 120 s, group G2 150 s. Treated group C2-G2 with incubation for 5 minutes in dark place and exposed by a semiconductor laser with a wavelength of 532 nm. The photosensitizer concentration was 40 mg/L. The spot radius was 0.5 cm and the power density was 0.14 W/cm2. Obtained samples after all treatments. Used the tabbed colony counting method to detect the inhibition effect of various groups on the biofilm of oral mixed bacteria in rat. Used atomic absorption spectrophotometer to determine the amount of calcium dissolved in the biofilm of rats after photodynamic therapy. We used the above two methods to evaluate the inhibitory effect of HMME-PDT on the cariogenic bacteria in dental plaque biofilms of rats. Results: 1. The bacteriostatic effect of HMME-PDT on bacterial plaque in rat oral cavity was studied in different concentration of photosensitizer. The tablet colony count results showed with the increase of concentration of photosensitizer stress the bacteria inhibitive rate was increased. Compared to the negative control group and positive control group, group E1 changed most obviously. 2. The bacteriostatic effect of HMME-PDT on bacterial plaque in rat oral cavity was studied in different exposure time. The tablet colony count results showed with the increase of exposure time the bacteria inhibitive rate was increased. Compared to the negative control group and positive control group, group E2 changed most obviously. 3. Results of the calcium ion dissolution detected by atomic absorption spectrophotometer showed with the increase of exposure time the calcium ion dissolution was increased in each group in 48 hours. Compared to the negative control group, the calcium ion dissolution of each group was decreased significantly (P<0.05). Compared to the positive control group, the calcium ion dissolution of each group was decreased significantly (P<0.05). Among all the groups group E1 and E2 changed most obviously. Conclusion: HMME-PDT has a significant inhibitory effect on the cariogenic bacteria in dental plaque biofilms of rats. The experiments proves that the optimal photosensitizer concentration of HMME-PDT was 40 mg/L and the optimal time of exposure duration was 90 s. Photodynamic therapy can effectively inhibit the production of caries in the biofilm of rats and reduce calcium ions solubility in early caries. PDT not only prevents the enamel from going off, but also promotes the remineralization of enamel. HMME-PDT has a broad clinical application prospect in the prevention of caries.

陶亚东, 柳雪, 霍峰, 王敬. HMME-PDT对大鼠口腔混合菌生物膜抑制作用的实验研究[J]. 应用激光, 2018, 38(1): 154. Tao Yadong, Liu Xue, Huo Feng, Wang Jing. The Experimental Study of the Effect of HMME-PDT Inhibition the Cariogenic Bacteria in Dental Plaque Biofilm of Rats[J]. APPLIED LASER, 2018, 38(1): 154.

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