血红素浓度对双菌生物膜中牙龈卟啉单胞菌致病潜力的影响

口腔生物医学 ›› 2021, Vol. 12 ›› Issue (2): 105-109.

血红素浓度对双菌生物膜中牙龈卟啉单胞菌致病潜力的影响

郭艳阳¹,王昱¹,顾宁²,汪宜瑾¹,靳雅冰¹,王琛¹

1. 南京医科大学附属口腔医院
2. 南京医科大学口腔医学院

收稿日期:2021-02-03 修回日期:2021-05-25 出版日期:2021-06-25 发布日期:2021-06-25
通讯作者: 王琛 E-mail:chwang9122@sina.com
基金资助:
国家自然科学基金;江苏省自然基金项目;江苏省高等学校优先学科建设项目;江苏省省级干部健康研究

Effects of heme concentration on pathogenicity of Porphyromonas gingivalis in the dual-species biofilm

Received:2021-02-03 Revised:2021-05-25 Online:2021-06-25 Published:2021-06-25
Contact: Chen -WANG E-mail:chwang9122@sina.com
Supported by:
the National Nature Science Foundation;the Natural Science Foundation of Jiangsu Province;the Priority Academic Program Development of Jiangsu Higher Education Institutions;Provincial Cadre Health Research of Jiangsu Province

摘要/Abstract

摘要： 目的：研究血红素浓度对白色念珠菌（C. albicans）-牙龈卟啉单胞菌（P. gingivalis）双菌生物膜中P. gingivalis致病潜力的影响。方法：在0.1?μg/mL（低）和5?μg/mL（高）血红素培养条件下构建P. gingivalis-C. albicans双菌生物膜以及P. gingivalis单菌生物膜，激光共聚焦显微镜观察生物膜中微生物的存活状况，根据标准曲线测定培养物上清以及菌细胞破碎后上清中P. gingivalis的胰酶样蛋白酶（TLP）活性，K-B纸片琼脂扩散法检测各组中P. gingivalis的药物抵抗性。结果：低血红素培养条件下，与单菌生物膜比较，双菌生物膜中P. gingivalis的活菌数量和生物膜厚度增宽明显增加（P＜0.05），生物膜培养液上清和P. gingivalis菌体内TLP活性均高于单菌生物膜（P＜0.05），P. gingivalis对头孢唑啉和环丙沙星的耐药率也有所提高，对磺胺甲噁唑均敏感。高血红素培养条件下，双菌生物膜与单菌生物膜之间在生物膜活死比例、TLP活性及药物敏感性方面无明显差异，且其中TLP活性与对头孢唑啉和环丙沙星的抗性均低于低血红素培养条件下的生物膜。结论：低血红素培养条件下，双菌生物膜中的P. gingivalis致病潜力增加。

关键词: 牙龈卟啉单胞菌, 血红素, 白色念珠菌, 致病潜力, 牙周炎

Abstract: Objective:?To study the effects of heme concentration on the pathogenic potential of P. gingivalis in C. albicans - P. gingivalis dual-species biofilm. Methods:?P. gingivalis-C. albicans dual-species biofilm and P. gingivalis single-species biofilm were established in a low- and high- heme environment. The living and dying status of microorganisms in biofilms was observed by laser confocal microscope. Trypsin-like proteinase (TLP) activity of P. gingivalis in culture supernatant and cells was determined by standard curve. The drug resistance of P. gingivalis was detected by K-B disk agar diffusion method. Results:?Under the condition of low heme culture, the number of viable P. gingivalis in the dual-species biofilm was significantly higher than that in the single-species biofilm, and as well as the thickness of the dual-species biofilm was wider in 3D images. The TLP activity in the supernatant and cell of P. gingivalis in dual-species biofilm was higher than that in single-species biofilm (P<0.05). In addition, the resistance rates of P. gingivalis in the dual-species biofilm against cefazolin and ciprofloxacin were also increased, while they were sensitive to sulfamethoxazole. There was no significant differences in Live/Death rate, TLP activity and drug sensitivity between the dual- and single-species biofilm under high heme culture condition, but the TLP activity and the resistance against cefazolin and ciprofloxacin were lower than those of the biofilms cultured in a low-heme environment. Conclusions:?Under the condition of low heme culture, the pathogenicity potential of P. gingivalis in the dual-species biofilm was increased.

Key words: Porphyromonas gingivalis, heme, Candida albicans, pathogenicity potential, periodontitis

郭艳阳王昱顾宁汪宜瑾靳雅冰王琛. 血红素浓度对双菌生物膜中牙龈卟啉单胞菌致病潜力的影响[J]. 口腔生物医学, 2021, 12(2): 105-109.

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