氧化银钛纳米管治疗骨关节假体相关性感染的实验研究

Experimental study on treatment of periprosthetic infection after joint replacement with titanium nanotubes loaded with nano silver oxide particles

目的探讨氧化银钛纳米管(Ag2O-TiO2-NTS)在治疗骨关节假体相关性感染过程中,对表面葡萄球菌及金黄色葡萄球菌的抑菌作用。方法2018年5月至2019年5月,从河北大学医学院实验动物中心购买健康新西兰大白兔32只。体外实验:选取不同浓度的Ag2O-TiO2-NTS溶液与表皮葡萄球菌及金黄色葡萄球菌共培养,并计算菌落数及抗菌率。通过细胞毒性和细胞增殖实验检测Ag2O-TiO2-NTS的生物学测定。体内实验:选用32只新西兰大白兔建立人工膝关节置换术后感染模型,并随机分为表-Ag钉组、表-普钉组、金-Ag钉组、金-普钉组。表-Ag钉组、金-Ag钉组兔子术中使用Ag2O-TiO2-NTS复合涂层的螺钉,表-普钉组、金-普钉组使用没有涂层的普通螺钉。拆线后1 d,于表-Ag钉组、表-普钉组兔术侧膝关节注入表皮葡萄球菌悬液,于金-Ag钉组、金-普钉组兔术侧膝关节注入金黄色葡萄球菌悬液。在手术当天、注射菌液当天及注射后1、3、5、7、10、14 d记录所有大白兔肛温、C反应蛋白(C-reactive protein,CRP)、红细胞沉降率(erythrocyte sedimentation rate,ESR)及降钙素原(procalcitonin,PCT)。结果不同浓度的Ag2O-TiO2-NTS溶液菌落数及抗菌率两两比较,仅1 mmol/L和2 mmol/L两组P>0.05,其余5 mmol/L组和10 mmol/L组两两比较,差异有统计学意义(P<0.05)。随着Ag2O-TiO2-NTS溶液浓度的增加,抗菌率逐渐增加,10 mmol/L Ag2O-TiO2-NTS溶液抗菌效果最强(32.34±1.75)CFU。表-Ag钉组、表-普钉组及金-Ag钉组和金-普钉组细胞死亡数目和细胞增殖数目差异无统计学意义(P>0.05),说明Ag2O-TiO2-NTS没有毒性且不影响细胞增殖,安全可靠。金-普钉组肛温注射后10 d最高(41.94±0.87)℃;金-普钉组CRP注射后14 d最高(91.95±6.47)mg/L;金-普钉组ESR注射后14 d最高(70.13±4.47)mg/L;金-普钉组PCT注射后14 d最高(2.58±0.54)μg/L,差异有统计学意义(P<0.05)。表-Ag钉组与表-普钉组比较及金-Ag钉组与金-普钉组比较,并发症发生率和死亡率,差异有统计学意义(P<0.05),金-普钉组并发症发生率为53.33%、死亡率为30.00%。结论Ag2O-TiO2-NTS在体内外均对表皮葡萄球菌及金黄色葡萄球菌具有良好的抗菌作用,在治疗骨关节假体相关性感染有良好前景。

Objective To investigate the antibacterial effect of titanium-based nanotubes(Ag2O-TiO2-NTS)loaded with nano-silver oxide particles on Staphylococcus epidermidis and Staphylococcus aureus in the treatment of peri-implant infection after artificial joint replacement.Methods From May 2018 to May 2019,32 healthy New Zealand white rabbits were purchased from the Laboratory Animal Center of Medical College of Hebei University.Ag2O-TiO2-NTS solutions with different concentrations were cocultured with Staphylococcus epidermidis and Staphylococcus aureus,and the number of colonies and antibacterial rate were calculated.Biological assays for Ag2O-TiO2-NTS were tested by cytotoxicity and cell proliferation assays.In vivo experiment:32 New Zealand white rabbits were used to establish an artificial joint replacement infection model,and were randomly divided into table-Ag nail group,table-nail group,gold-Ag nail group and gold-nail group.Ag2O-TiO2-NTS composite coating screws were used in rabbits in table-Ag nail group and gold-Ag nail group,while ordinary screws without coating were used in the table-nail group and goldnail group.One day after the suture removal,Staphylococcus epidermidis suspension was injected into the knee joint of the table-Ag nail group and the table-nail group,and Staphylococcus aureus suspension was injected into the knee joint of the gold-Ag nail group and the gold-nail group.Anal temperature,CRP,ESR and PCT were recorded on the day of surgery,on the day of injection of bacteria and on days 1,3,5,7,10 and 14 after injection.Results The colony count and antibacterial rate of Ag2O-TiO2-NTS solution with different concentrations were compared,only 1 mmol/L and 2 mmol/L groups(P>0.05).The other 5 mmol/l group and 10 mmol/l group showed significant difference(P<0.05).With the increase of the concentration of Ag2O-TiO2-NT solution,the antibacterial rate increased gradually.The antibacterial effect of 10 mmol/L Ag2O-TiO2-NT solution was the strongest(32.34±1.75)CFU.There was no significant difference in the number of cell death and cell proliferation between the two groups(P>0.05),indicating that Ag2O-TiO2-NT had no toxicity and did not affect cell proliferation,which was safe and reliable.The highest anal temperature was(41.94±0.87)℃in the gold-nail group 10 days after injection;the highest CRP was(91.95±6.47)mg/L at 14 d after injection;ESR was the highest at 14 days after ESR injection,which was(70.13±4.47).There were statistically significant differences in the incidence of complications and mortality between the two groups(P<0.05).The incidence of complications and mortality in the gold silver nail group were 53.33%and 30.00%,respectively.Conclusion Ag2O-TiO2-NTS has good antibacterial activity against Staphylococcus epidermidis and Staphylococcus aureus in vitro and in vivo,and has a good prospect in the treatment of periprosthetic infection after artificial joint replacement.