摘要
目的探究低频超声对关节假体周围感染(PJI)术中常用冲洗液在体外模型中清除金黄色葡萄球菌生物膜及杀菌的增效作用。方法实验时间为2023年10月至2024年8月。在钛片表面构建48 h金葡菌生物膜模型,将36枚钛片按随机数字表法分为12组:0.9%氯化钠对照组、0.9%氯化钠超声10 s组、0.9%氯化钠超声5 min组、3%过氧化氢对照组、3%过氧化氢超声10 s组、3%过氧化氢超声5 min组、0.1%苯扎氯铵对照组、0.1%苯扎氯铵超声10 s组、0.1%苯扎氯铵超声5 min组、0.5%聚维酮碘对照组、0.5%聚维酮碘超声10 s组和0.5%聚维酮碘超声5 min组,每组3枚钛片,分别置于30 ml冲洗液中进行干预,干预后的钛片再行细菌活/死染色和扫描电镜观察;钛片再培养,检测残留菌的活性。两组间比较用t检验,多组间比较采用方差分析One-way和Two-way ANOVA方法检验,进一步两两比较用Tukey法进行检验。结果生物膜活/死细胞染色、扫描电镜及钛片再培养结果显示4种冲洗液仅浸泡5 min不能杀灭生物膜中残留细菌;生物膜活/死细胞染色残留生物膜面积定量分析结果中,0.9%氯化钠组:超声5 min组显著低于超声10 s组[(0.092±0.087)%比(0.477±0.079)%;t=6.969,P<0.01],超声10 s组和超声5 min组显著低于对照组[(0.477±0.079)%比100%、(0.092±0.087)%比100%,t=1801.000、1808.000,P<0.01];0.5%聚维酮碘组:超声5 min组和超声10 s组差异无统计学意义[(0.041±0.068)%比(2.717±2.365)%,t=2.400,P>0.05],超声10 s组和超声5 min组显著低于对照组[(2.717±2.365)%比100%、(0.041±0.068)%比100%,t=87.230、89.630,P<0.01];3%过氧化氢组:超声10 s组和超声5 min组差异无统计学意义[(0.154±0.156)%比(0.060±0.054)%,t=1.203,P>0.05],超声10 s组和超声5 min组显著低于对照组[(0.154±0.156)%比100%、(0.060±0.054)%比100%,t=1281.000、1282.000,P<0.01];0.1%苯扎氯铵组:超声5 min组低于超声10 s组[(0.040±0.012)%比(0.473±0.202)%,t=4.523,P<0.05],超声10 s组和超声5 min组显著低于对照组[(0.473±0.202)%比100%、(0.040±0.012)%比100%,t=1040.000、1044.000,P<0.01]。4种冲洗液组间比较,超声10 s时,0.5%聚维酮碘组显著高于0.9%氯化钠组、3%过氧化氢组、0.1%苯扎氯铵组[(2.717±2.365)%比(0.477±0.079)%、(0.154±0.156)%、(0.473±0.202)%,q=4.605、5.269、4.614,P<0.05];在超声5 min时,0.9%氯化钠、3%过氧化氢、0.1%苯扎氯铵和0.5%聚维酮碘4种冲洗液之间差异无统计学意义[(0.092±0.087)%比(0.060±0.054)%比(0.041±0.068)%比(0.040±0.012)%,P>0.05]。结论低频超声联合0.9%氯化钠可显著清除钛片表面生物膜,而低频超声联合0.5%聚维酮碘或0.1%苯扎氯铵可有效清除生物膜并杀灭残留菌。
Objective To investigate the potentiating effect of low-frequency ultrasound on the removal of Staphylococcus aureus(S.aureus)biofilm and sterilization by commonly used irrigation solutions in periprosthetic joint infection(PJI)surgery in an in vitro model.Methods The experimental period is from October 2023 to August 2024.A 48-hours.aureus biofilm model was constructed on the surface of titanium plates,and 36 titanium plates were divided into 12 groups according to the randomized numerical table method:0.9%sodium chloride control group,0.9%sodium chloride ultrasound 10 s group,0.9%sodium chloride ultrasound 5 min group,3%hydrogen peroxide control group,3%hydrogen peroxide ultrasound 10 s group,3%hydrogen peroxide ultrasound 5 min group,and 0.1%benzalkonium chloride control group,0.1%benzalkonium chloride ultrasound 10 s group,0.1%benzalkonium chloride ultrasound 5 min group,0.5%povidone-iodine control group,0.5%povidone-iodine ultrasound 10 s group,and 0.5%povidone-iodine ultrasound 5 min group,3 titanium plates in each group,respectively,were placed in the 30 ml of irrigation solution for the intervention,and then the titanium plates were subjected to live/dead staining of the bacteria and scanning electron microscopy observation after the intervention;the titanium plates were re-cultured to detect the activity of residual bacteria.Data were analyzed using t-tests for pairwise comparisons,One-way and Two-way ANOVA for multiple comparisons,and Tukey’s test for post-hoc analyses.Results The results of biofilm live/dead cell staining,scanning electron microscopy(SEM),and titanium piece re-culture showed that the four irrigation solutions could not kill the residual bacteria in the biofilm by only 5 min immersion;in the results of quantitative analysis of the biofilm area of residual biofilm in biofilm live/dead cell staining,the results of the 0.9%sodium chloride group:the group of ultrasound for 5 min was significantly lower than that of the group of ultrasound for 10 s[(0.092±0.087)%vs.(0.477±0.079)%,t=6.969,P<0.01],and significantly lower in the ultrasound 10 s and ultrasound 5 min groups than in the control group[(0.477±0.079)%vs.100%,(0.092±0.087)%vs.100%,t=1801.000,1808.000,P<0.01];0.5%povidone-iodine group:ultrasound 5 min and ultrasound 10 s groups were not significantly different[(0.041±0.068)%vs.(2.717±2.365)%,t=2.400,P>0.05],and ultrasound 10 s and ultrasound 5 min groups were significantly lower than the control group[(2.717±2.365)%vs.100%,(0.041±0.068)%vs.100%,t=87.230,89.630,P<0.01];3%hydrogen peroxide group:there was no significant difference between the ultrasound 10 s and ultrasound 5 min groups[(0.154±0.156)%vs.(0.060±0.054)%,t=1.203,P>0.05],and the ultrasound 10 s and ultrasound 5 min groups were significantly lower than the control group[(0.154±0.156)%vs.100%,(0.060±0.054)%vs.100%,t=1281.000,1282.000,P<0.01];0.1%benzalkonium chloride group:ultrasound 5min group was lower than ultrasound 10s group[(0.040±0.012)%vs.(0.473±0.202)%,t=4.523,P<0.05],and significantly lower in the ultrasound 10 s and ultrasound 5 min groups than in the control group[(0.473±0.202)%vs.100%,(0.040±0.012)%vs.100%,t=1040.000,1044.000,P<0.01].Comparing among the four irrigation solution groups,at 10 s of ultrasound,the 0.5%povidone-iodine group was significantly higher than the 0.9%sodium chloride group,the 3%hydrogen peroxide group,and the 0.1%benzalkonium chloride group[(2.717±2.365)%vs.(0.477±0.079)%,(0.154±0.156)%,(0.473±0.202)%,q=4.605,5.269,4.614,P<0.05];at 5 min of ultrasound,there was no significant difference between the four irrigation solutions,0.9%sodium chloride,3%hydrogen peroxide,0.1%benzalkonium chloride,and 0.5%povidone-iodine[(0.092±0.087)%vs.(0.060±0.054)%vs.(0.041±0.068)%vs.(0.040±0.012)%,P>0.05].Conclusion Low-frequency ultrasound combined with 0.9%sodium chloride significantly removes biofilms on titanium surfaces,while its combination with 0.5%povidone-iodine or 0.1%benzalkonium chloride effectively eliminates biofilms and kills residual bacteria.
作者
孟兴补
李亦丞
汪斐
肖扬
曹力
Meng Xingbu;Li Yicheng;Wang Fei;Xiao Yang;Cao Li(Department of Orthopedics,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)
出处
《中华实验外科杂志》
2025年第4期701-706,共6页
Chinese Journal of Experimental Surgery
基金
新疆维吾尔自治区科技计划重大专项项目(2022A03011)
新疆维吾尔自治区科学技术厅科技创新团队项目(2023TSYCTD0014)
新疆维吾尔自治区自然科学基金(2023D01E11)。
关键词
关节假体周围感染
金黄色葡萄球菌
生物膜
冲洗液
低频超声
Periprosthetic joint infection
Staphylococcus aureus
Biofilm
Irrigation solution
Low-Frequency ultrasound
