健康志愿者自体富血小板凝胶体外抑菌作用研究

ANTIBACTERIAL EFFECT OF AUTOLOGOUS PLATELET-RICH GEL DERIVED FROM HEALTH VOLUNTEERS IN VITRO

目的自体富血小板凝胶(autologous platelet-rich gel,APG)作为一种新技术和有希望广泛用于临床的抗感染治疗方法,其作用机制尚不清楚。许多研究表明血小板具有抗菌的多功能属性,可直接与病原微生物作用将病原体从血液中清除。通过体外实验评价APG是否具有抑菌作用。方法抽取17例健康志愿者外周静脉血,制备富血小板血浆(platelet-rich plasma,PRP)、贫血小板血浆(platelet-poor plasma,PPP)及APG。通过抑菌实验观察APG、PRP以及加入夹竹桃麻素(apocynin,APO)的APG(APG-APO)对金黄色葡萄球菌、大肠埃希杆菌及铜绿假单胞菌是否有抑菌作用。结果APG和APG-APO对金黄色葡萄球菌的生长具有明显抑制作用,最大抑制作用出现在前4h,抑菌作用分别可持续至24h和8h。APG与PRP、PPP相比,24h内均具有显著的抑菌作用(P<0.05);在前8h APG与APG-APO的抑菌作用差异无统计学意义(P>0.05),12h后差异有统计学意义(P<0.05);APG-APO与PRP、PPP相比在前4h具有显著的抑菌作用(P<0.05),6h后差异无统计学意义(P>0.05)。各时间点APG和APG-APO对大肠埃希杆菌的抑菌率分别为27.36%～52.97%和18.82%～51.52%,对铜绿假单胞菌的抑菌率均在35%以下,与PRP比较差异均无统计学意义(P>0.05);各时间点PRP对金黄色葡萄球菌、大肠埃希杆菌及铜绿假单胞菌的抑菌率均在15%以下。结论APG对金黄色葡萄球菌有抑菌作用,抑菌作用可能来自于血小板;对大肠埃希杆菌及铜绿假单胞菌无明显抑菌作用。PRP对金黄色葡萄球菌、大肠埃希杆菌及铜绿假单胞菌均无明显抑菌作用。

Objective The use of autologous platelet-rich gel （APG） is a relatively new technology and a promising treatment method for infections,which is currently being used by a variety of surgical specialties. The mechanism of antibacterial effect of APG is not yet fully discovered. Subsequent evidence suggests that platelets have multiple functional attributes in antimicrobial host defense （including the capacity to generate antimicrobial oxygen metabolites and the antimicrobial peptides） and interact directly with microorganisms,contribute to clearance of pathogens from the blood. To investigate the bacteriostasis of APG against Staphylococcus aureus,Escherichia coli,and Pseudomonas aeruginosa in vitro. Methods Platelet-rich plasma （PRP） and platelet-poor plasma （PPP） were obtained from whole blood of 17 healthy donors. APG was prepared by mixing PRP with bovine thrombin in a 10% calcium gluconate solution or bovine thrombin in a 10% calcium gluconate solution and apocynin （APG-APO）. Antibacterial effects of APG,PRP,and APG-APO on Staphylococcus aureus,Escherichia coli,and Pseudomonas aeruginosa were evaluated by bacteriostasis assay. Results The culture results showed apparent decrease in the number of Staphylococcus aureus for both APG and APG-APO,which was maximal at first 4 hours and lasted to 24 hours and 8 hours,respectively; showing significant difference （P〈 0.05） when compared APG with PRP and PPP,however no signifi cant difference at first 8 hours （P 〉0.05） and significant difference at 12 and 24 hours （P 〈0.05） when compared APG with APG-APO; showing significant difference at first 4 hours （P〈 0.05）,no significant difference at 6,8,12,and 24 hours when compared APG-APO with PRP and PPP （P 〉0.05）. The bacteriostasis rates of APG and APG-APO were 27.36%-52.97% and 18.82%-51.52% against Escherichia coli,respectively; showing no significant difference （P 〉0.05） when compared with PRP.The bacteriostasis rates of APG and APG-APO were less than 35% against Pseudomonas aeruginosa,showing no significant difference （P 〉0.05） when compared with PRP; the bacteriostasis rates of PRP were less than 15% against Staphylococcus aureus,Escherichia coli,and Pseudomonas aeruginosa. Conclusion APG may have potential bacteriostatic effect against Staphylococcus aureus by platelet mediating. Either APG or APG-APO has no obvious bacteriostatic effect against Escherichia coli or Pseudomonas aeruginosa. PRP has no antibacterial activity against Staphylococcus aureus,Escherichia coli or Pseudomonas aeruginosa.