一氧化氮缓释二氧化硅纳米颗粒对内镜生物膜的清除效果及其临床应用评价

Cleansing effect of NO sustained⁃release silica nanoparticles on endoscopic biofilm and its clinical application

目的探讨一氧化氮(Nitric Oxide,NO)缓释二氧化硅纳米颗粒(简称NO缓释剂)对内镜生物膜的清除效果及其临床应用评价。方法选择院内临床使用中的消化内镜160件,随机分为两组:清洗剂组80件,采用清洗剂对内镜进行消毒处理;NO组80件,采用NO缓释剂对内镜进行消毒处理。另建模型组,采用铜绿假单胞菌生物膜模型,用磷酸盐缓冲液(phosphate buffered solution,PBS)处理,作为对照。先在体外层面,构建内镜管腔铜绿假单胞菌生物膜模型,经NO缓释剂处理后采用扫描电镜观察生物膜微结构,采用活菌计数法分析NO缓释剂对生物膜细菌的清除效果,再在临床层面,通过分析清洗后临床内镜的蛋白残留测定、菌落计数及ATP生物荧光检测评价NO缓释剂对内镜的实际消毒效果。结果扫描电镜观察,模型组生物膜完整,NO组和清洗剂组生物膜表面可见散落杆菌。平均标准菌落数,与模型组[(4.86±2.67)×10^(6)菌落形成单位(colony⁃forming units,CFU)/mL]比,NO组[(1.37±0.61)×10^(4)CFU/mL]和清洗剂组[(1.31±0.21)×10^(5)CFU/mL]均显著降低(清洗剂组比模型组,P=0.009;NO组比模型组,P=0.008),且NO组比清洗剂组更低,组间差异有统计学意义(清洗剂组比NO组,t=9.53,P=0.0006)。临床消化内镜层面,残留蛋白,NO组处理前后分别为(8.03±1.47)mg/mL、(0.50±0.37)mg/mL,清洗剂组处理前后分别为(8.01±1.51)mg/mL、(0.91±0.52)mg/mL,各组处理前后比较差异有统计学意义(P<0.01),且NO组降低幅度更大。临床内镜分别经NO缓释剂与清洗剂处理,NO组与清洗剂组的消毒在合格范围内,但NO组的ATP生物荧光值[(78.56±42.59)RLU]、蛋白残留量[(0.50±0.37)mg/mL]与菌落计数量[(7.55±4.56)CFU]均显著低于清洗剂组[(120.80±54.00)RLU,(0.91±0.52)mg/mL,(11.50±4.75)CFU],差异均有统计学意义(P均<0.01)。结论NO缓释剂能够有效清除内镜生物膜,提高内镜消毒效果,可能对改善患者的诊疗效果及预后具有重要意义。

Objective To explore the cleansing effect of Nitric Oxide(NO)sustained-release silica nanoparticles(short for NO sustained-release agent)on endoscopic biofilm and its clinical application.Methods A total of 160 clinical endoscopes were randomly divided into two groups:the cleansing agent group(80 pieces,disinfected with cleansing agents),NO group(80 pieces,disinfected with NO sustained-release agent).A biofilm model of Pseudomonas aeruginosa was constructed and used as the control for phosphate buffered solution(PBS)treatment.A biofilm model of Pseudomonas aeruginosa on the surface of endoscopic lumen was built first in vitro.Scanning electron microscopy was then used to observe the microstructure of biofilm after treatment with NO sustained-release agent.Viable counting method was used to evaluate the cleansing effect of NO sustained-release agent on biofilm.Finally,at the clinical level,the actual disinfection effect of NO sustained-release agent on clinical endoscopy was evaluated by detecting the protein residues,viable counting and adenosine triphosphate(ATP)biofluorescence detection.Results The scanning electron microscopy showed that the biofilm was intact in the model group,but scattered bacteria were observed on the biofilm surface in the NO group and the detergent group.Compared with the model group[(4.86±2.67)×10^(6)(colony-forming units,CFU)/mL],the standard CFUs of the NO group[(1.37±0.61)×10^(4)CFU/mL]and the detergent group[(1.31±0.21)×10^(5)CFU/mL]were significantly lower(detergent group VS model group,P=0.009;NO group VS model group,P=0.008),and there was significant difference between the detergent group and the model group(t=9.53,P=0.0006).The levels of residual proteins in the endoscopic lumens before and after the treatment were 8.03±1.47 mg/mL and 0.50±0.37 mg/mL in the NO group,8.01±1.51 mg/mL and 0.91±0.52 mg/mL in the detergent group with significant difference(P<0.01),and the reduction effect of the NO group was more significant.The disinfection of NO group and cleaning agent group was within the qualifying range,but the ATP bioluminescence value,protein residue and colony count of NO group(78.56±42.59 RLU,0.50±0.37 mg/mL,7.55±4.56 CFU)were significantly lower than those of detergent agent group(120.80±54.00 RLU,0.91±0.52 mg/mL,11.50±4.75 CFU,P<0.01).Conclusion NO sustained-release agent can effectively clear endoscopic biofilm and further improve the disinfection effect on endoscopes,which may be of great significance for improving the effects on treatment and prognosis of patients.