中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2010年
5期
411-415
,共5页
徐琼%黎卫兰%卢婷%刘正政
徐瓊%黎衛蘭%盧婷%劉正政
서경%려위란%로정%류정정
肠球菌,粪%生物膜%细胞外基质%牙本质%葡聚糖酶
腸毬菌,糞%生物膜%細胞外基質%牙本質%葡聚糖酶
장구균,분%생물막%세포외기질%아본질%포취당매
Enterococcus faecalis%Biofilm%Extracellular matrix%Dentin%Dextranase
目的 研究胞外多糖和胞外DNA对粪肠球菌在人牙本质片上黏附并形成生物膜过程中的作用.方法 制备牙本质片,分成葡聚糖酶(Dextranase)组、DNA酶(DNaseI)组和对照组,体外培养获得粪肠球菌的牙本质生物膜标本.采用活菌菌落计数法、激光共聚焦扫描显微镜结合COMSTAT软件分析胞外聚合物对牙本质黏附菌量和生物膜三维结构的影响.结果 相同生物膜膜龄,Dextranase组和DNaseI组粪肠球菌牙本质黏附菌量均少于对照组,差异具有统计学意义(P＜0.05),其中1 h黏附菌量分别下降至对照组的15.61%、18.39%.激光共聚焦显微镜观察到Dextranase和DNaseI对粪肠球菌牙本质生物膜的形成有影响,处理后生物膜结构松散,生物量、底层覆盖率及平均厚度均下降,具有统计学意义(P＜0.0167),其中生物量由(3.95±0.09)μm3/μm2下降至(0.36±0.08)μm3/μm2、(1.05±0.06)μm3/μm2.DNaseI对成熟牙本质生物膜产生显著降解作用,而Dextranase对成熟牙本质生物膜作用不明显.结论 胞外多糖和胞外DNA是粪肠球菌生物膜的重要成分,对其初始黏附和生物膜三维结构起重要作用.
目的 研究胞外多糖和胞外DNA對糞腸毬菌在人牙本質片上黏附併形成生物膜過程中的作用.方法 製備牙本質片,分成葡聚糖酶(Dextranase)組、DNA酶(DNaseI)組和對照組,體外培養穫得糞腸毬菌的牙本質生物膜標本.採用活菌菌落計數法、激光共聚焦掃描顯微鏡結閤COMSTAT軟件分析胞外聚閤物對牙本質黏附菌量和生物膜三維結構的影響.結果 相同生物膜膜齡,Dextranase組和DNaseI組糞腸毬菌牙本質黏附菌量均少于對照組,差異具有統計學意義(P＜0.05),其中1 h黏附菌量分彆下降至對照組的15.61%、18.39%.激光共聚焦顯微鏡觀察到Dextranase和DNaseI對糞腸毬菌牙本質生物膜的形成有影響,處理後生物膜結構鬆散,生物量、底層覆蓋率及平均厚度均下降,具有統計學意義(P＜0.0167),其中生物量由(3.95±0.09)μm3/μm2下降至(0.36±0.08)μm3/μm2、(1.05±0.06)μm3/μm2.DNaseI對成熟牙本質生物膜產生顯著降解作用,而Dextranase對成熟牙本質生物膜作用不明顯.結論 胞外多糖和胞外DNA是糞腸毬菌生物膜的重要成分,對其初始黏附和生物膜三維結構起重要作用.
목적 연구포외다당화포외DNA대분장구균재인아본질편상점부병형성생물막과정중적작용.방법 제비아본질편,분성포취당매(Dextranase)조、DNA매(DNaseI)조화대조조,체외배양획득분장구균적아본질생물막표본.채용활균균락계수법、격광공취초소묘현미경결합COMSTAT연건분석포외취합물대아본질점부균량화생물막삼유결구적영향.결과 상동생물막막령,Dextranase조화DNaseI조분장구균아본질점부균량균소우대조조,차이구유통계학의의(P＜0.05),기중1 h점부균량분별하강지대조조적15.61%、18.39%.격광공취초현미경관찰도Dextranase화DNaseI대분장구균아본질생물막적형성유영향,처리후생물막결구송산,생물량、저층복개솔급평균후도균하강,구유통계학의의(P＜0.0167),기중생물량유(3.95±0.09)μm3/μm2하강지(0.36±0.08)μm3/μm2、(1.05±0.06)μm3/μm2.DNaseI대성숙아본질생물막산생현저강해작용,이Dextranase대성숙아본질생물막작용불명현.결론 포외다당화포외DNA시분장구균생물막적중요성분,대기초시점부화생물막삼유결구기중요작용.
Objective To investigate the effects of extracellular polysaccharide and extracellular DNA on adhesion and biofilm formation of Enterococcus faecalis in ground sections of human dentin.Methods Ground sections of dentin were prepared and allocated to three groups: Dextranase group, DNaseI group and normal control group. Biofilm samples of Enterococcusfaecalis on dentin were obtained by culture in vitro. Colony-forming units (CFU) counts, confocal laser scanning microscope (CLSM) and COMSTAT software were used to measure the effect of extracellular polysaccharide on the bacterial adherence to dentin and the three dimensional structure of biofilm. Results Given a certain age of biofilm, less bacterial adherence of Enterococcus faecalis to dentin was found in both Dextranase and DNaseI group than that in normal control group (P＜0.05). The 1-h bacterial adherence in these two groups was 15.61% and 18.39% less, respectively, as compared with the normal control group. CLSM showed that treatment with Dextranase or DNaseI may affected dentin biofilm formation: loosened biofilm structure and reduction in biomass, bottom coverage and mean thickness (P＜0.0167). Notably, the biomass in Dextranase and DNaseI group decreased significantly from ( 3.95 ± 0.09 ) μm3/μm2 to (0.36 ± 0.08 ) μm3/μm2 and ( 1.05 ± 0.06 ) μm3/μm2, respectively.DNaseI but not Dextranase was shown to degrade the mature biofilms on dentin. Conclusion Extracellular polysaccharide and extracellular DNA may be important components of Enterococcus faecalis bioflim structure, which contribute significantly to initial bacterial adhesion and three dimensional biofilm structure.
