亚甲基蓝介导的光动力疗法联合小檗碱对牙龈卟啉单胞菌抑制作用的体外研究  

Inhibitory effect of methylene blue mediated photodynamic therapy combined with berberine on

作　　者：李天甜 闫诗雯 江河 王梦雯 潘文婷[1] 薛鹏[1] Li Tiantian;Yan Shiwen;Jiang He;Wang Mengwen;Pan Wenting;Xue Peng(The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学第一附属医院,郑州450000

出　　处：《国际生物医学工程杂志》2023年第1期23-29,共7页International Journal of Biomedical Engineering

基　　金：河南省医学科技攻关省部共建重点项目(SBGJ202102159);河南省高等学校重点科研项目(20A320082);中国工程科技发展战略河南省研究院咨询研究项目(2022HENYB02);河南省医学科技攻关联合共建项目(71166)

摘　　要：目的探讨亚甲基蓝介导的光动力疗法(PDT)联合小檗碱对牙龈卟啉单胞菌(P.g)的体外抑制作用。方法培养P.g至对数期中后期,将不同质量浓度亚甲基蓝加入P.g菌悬液中,作用5 min,激光(波长660 nm,功率140 mW/cm^(2))照射2 min,寻找亚甲基蓝结合激光体外抑制P.g的最佳浓度;观察亚甲基蓝介导的PDT体外抑制P.g的效果以及小檗碱对P.g生长曲线的影响;探讨亚甲基蓝介导的PDT与小檗碱先后联合应用对P.g的抑制作用;扫描电子显微镜观察亚甲基蓝介导的PDT及小檗碱对P.g形态的影响;紫外分光光度仪测量各成分吸收峰。结果在660 nm激光激发下,亚甲基蓝质量浓度为24.4141μg/ml时,抑菌效果最好。与对照组比较,亚甲基蓝组和PDT组差异均有统计学意义(均P<0.001)。0.05 mg/ml小檗碱对P.g浮游细菌具有抑制作用。与对照组比较,0.05 mg/ml小檗碱组菌落数降低,其差异有统计学意义(P<0.01);0.05 mg/ml小檗碱+光照组与对照组比较差异无统计学意义(P>0.05)。当PDT与小檗碱联用时对P.g有协同抑制作用,且先PDT后小檗碱组较先小檗碱后PDT组菌落数降低,其差异均有统计学意义(均P<0.01)。P.g经亚甲基蓝介导的PDT处理后,细菌细胞壁皱缩成团,经小檗碱处理后,细菌表面变得光滑且菌体长度较对照组增长。结论亚甲基蓝介导的PDT对P.g有抑制作用,当与小檗碱联用时,对P.g有协同抑制作用,且联合应用中先PDT后小檗碱作用时抑菌效果更好。Objective To investigate the in vitro inhibitory effect of methylene blue mediated photodynamic therapy(PDT)combined with berberine on Porphyromonas gingivalis(P.g).Methods P.g was cultured until the middle to late log phase,and methylene blue was added to P.g suspension at different mass concentrations for 5 min,and a laser(wavelength 660 nm,power 140 mW/cm^(2))was irradiated for 2 min to find the optimal concentration of methylene blue combined with the laser for in vitro inhibition of P.g.The effect of methylene blue mediated PDT on the in vitro inhibition of P.g and the effect of berberine on the growth curve of P.g were observed.The inhibitory effect of methylene blue mediated PDT and berberine on P.g was investigated by successive combined applications.The effect of methylene blue mediated PDT on P.g morphology was observed by scanning electron microscopy.The absorption peaks of each component were measured by ultraviolet spectrophotometer.Results The best inhibition was achieved at a methylene blue mass concentration of 24.4141μg/ml under 660 nm laser excitation.The differences were statistically significant in both the methylene blue and PDT groups compared with the control group(all P<0.001).0.05 mg/ml berberine had an inhibitory effect on the planktonic bacteria of P.g.After P.g was treated with methylene blue mediated PDT,the bacterial cell walls were crumpled into clusters.Compared with the control group,the number of colonies was reduced in the 0.05 mg/ml berberine group,and the difference was statistically significant(P<0.01).The difference between the 0.05 mg/ml berberine+light group and the control group was not statistically significant(P>0.05).When PDT was combined with berberine,there was a synergistic inhibitory effect on P.g.PDT followed by berberine shows a better inhibitory effect on bacteria,and the differences were statistically significant(all P<0.01).After the berberine treatment,the bacterial surface became smooth,and the length of the bacterial body increased compared with the contro

关 键 词：亚甲基蓝 光动力疗法 小檗碱 牙龈卟啉单胞菌 抑菌作用 

分 类 号：R781.4[医药卫生—口腔医学]