不同方法制备的富血小板血浆对烧伤感染创面常见致病菌的体外抑菌实验

doi:10.3877/cma.j.issn.2095-5820.2025.02.004

目的

通过体外抑菌实验探究不同制备方法获得的富血小板血浆（PRP）及富血小板凝胶上清（PGS）对烧伤创面常见致病菌金黄色葡萄球菌（SA）、表皮葡萄球菌（SE）及铜绿假单胞菌（PA）的体外抑菌效应。

方法

获取SA、SE及PA临床菌株，分别使用机采法和手工二次离心法采集健康志愿者新鲜全血制备PRP，同时将部分手工法获得的PRP制备成PGS，采用打孔法抑菌实验，以PRP和PGS为实验组，以灭菌纯水为阴性对照组，观察各组是否产生抑菌圈并测量其直径。

结果

手工二次离心法制备的PRP在SA和PA组可产生明显抑菌圈，平均抑菌圈直径为（2.164±0.354）mm和（3.544±0.507）mm，而在SE组未见明显抑菌圈；机采法制备的PRP组及手工法制备的PGS在3种细菌培养皿中均未产生抑菌圈。

结论

手工二次离心法制备的PRP对烧伤创面常见致病菌SA和PA具有抑菌效应，但对SE无抑菌效应；机采法制备的PRP及手工二次离心法制备的PGS对这3种烧伤创面常见致病菌均无抑菌效应。

关键词: 机采法, 手工二次离心法, 富血小板血浆, 烧伤创面, 感染, 抑菌实验

Objective

To investigate the vitro antibacterial effects of platelet-rich plasma (PRP)and platelet-rich gel supernant (PGS), prepared using different methods against common burn wound pathogens: Staphylococcus epidermidis (SE), Staphylococcus aureus (SA) and Pseudomonas aeruginosa (PA).

Methods

Clinical isolates of SA, SE, and PA were collected, and PRP was collected from healthy volunteers by mechanical collection method and manual two-step centrifugation method, and part of PRP obtained by manual two-step centrifugation method was prepared into PGS. Oxford cup assay method was used to observe the inhibition zone of PRP, PGS and control group and determine its diameter.

Results

The PRP prepared by manual two-step centrifugation method demonstrated significant antibacterial activity against SA(2.164±0.354 mm inhibitory zone) and PA (3.544±0.507 mm inhibitory zone), but no obvious inhibitory zone was observed in SE group. Neither apheresis-prepared PRP nor manual PGS showed detectable inhibition zones against any tested pathogens.

Conclusions

This study demonstrated that PRP prepared by manual two-step centrifugation method has antibacterial effect on SA and PA, but has no antibacterial effect on SE. PRP prepared by mechanical collection method and PGS prepared by manual two-step centrifugation method lack measurable antimicrobial activity against these three common burn wound pathogens.

Key words: mechanical collection method, manual two-step centrifugation method, plateletrich plasma, burn wound, infection, antibacterial effect

图1 SA、SE和PA在血平皿中菌落形态注：1A为 SA，1B为SE，1C为PA。

表1 手工法和机采法制备PRP中PLT、WBC

项目	PLT/10⁹/L					WBC/10⁹/L
项目	A	B	C	D	均值/x̄ ± s	A	B	C	D	均值/x̄ ± s
手工法 PRP	900	924	970	995	947.3±46.54	0.48	0.54	0.60	0.69	0.578±0.078
机采法 PRP	1003	1010	989	970	993.0±17.64	0.01	0.01	0.01	0.00	0.008±0.005
P 值	-	-	-	-	0.092	-	-	-	-	<0.000 1

表1 手工法和机采法制备PRP中PLT、WBC

项目	PLT/10⁹/L					WBC/10⁹/L
项目	A	B	C	D	均值/x̄ ± s	A	B	C	D	均值/x̄ ± s
手工法 PRP	900	924	970	995	947.3±46.54	0.48	0.54	0.60	0.69	0.578±0.078
机采法 PRP	1003	1010	989	970	993.0±17.64	0.01	0.01	0.01	0.00	0.008±0.005
P 值	-	-	-	-	0.092	-	-	-	-	<0.000 1

图2 培养12 h后，SA、SE和PA 3种细菌抑菌圈产生情况注：2A为SA组，2B为SE组，2C为PA组；左上角“P1”代表手工法PRP组，右上角“P2”代表机采法PRP组，左下角“P3”代表PGS组，右下角“C”代表灭菌纯水组。

图3 培养12 h后，SA、SE和PA 3种细菌手工法PRP抑菌圈直径注：与SA比较，^aP＜0.000 1；与SE比较，^bP＜0.000 1。

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In vitro experimental study on the antibacterial effect of platelet-rich plasma prepared by different methods against common pathogenic bacteria in burn infected wound

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In vitro experimental study on the antibacterial effect of platelet-rich plasma prepared by different methods against common pathogenic bacteria in burn infected wound