1.400038 重庆, 陆军军医大学(第三军医大学)第一附属医院内分泌科
2.430070武汉, 解放军武汉总医院内分泌科
3.400041 重庆, 重庆市急救医疗中心内分泌科/糖尿病足诊治中心
Received:13 May 2018,
Revised:2018-09-10,
Published:15 December 2018
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Min HE, Tao LI, Min WANG, et al. Antibacterial role of platelet-rich plasma of different forms in
Min HE, Tao LI, Min WANG, et al. Antibacterial role of platelet-rich plasma of different forms in
目的
2
在体外高糖环境的感染创面模型中研究富血小板血浆(platelet-ric
h plasma
PRP)、富血小板凝胶(platelet-rich gel
PRG)及富血小板凝胶萃取液(extract liquid of platelet-rich gel
EPG)对金黄色葡萄球菌(staphylococcus aureus
S. aureus
)的抗菌作用
以及EPG对受细菌感染的人角质形成细胞(HaCaT细胞)的增殖作用。
方法
2
新鲜全血采集自PRG治疗的糖尿病足患者
离心后获得贫血小板血浆(platelet-poor plasma
PPP)和PRP。将HaCaT细胞与
S. aureus
在高糖环境下共培养建立体外感染创面模型。以PPP干预模型为对照组
试验组以干预模型分为PRP组、PRG组和EPG组(
n
=3)。通过平板菌落计数法对不同时间点各组中活菌进行计数
计数结果以log(CFU/mL)表示。以CCK-8法检测单纯细胞培养组
细胞与细菌共培养组及EPG干预组中HaCaT细胞的增殖情况。
结果
2
干预12、24 h PRP组和EPG组中HaCaT细胞外
S. aureus
计数均显著低于PPP组(
P
<
0.05);干预48 h PRG组和EPG组显著低于PPP组(
P
<
0.05)
而PRP组和PPP组
以及PRG组和EPG组差异均无统计学意义(
P
>
0.05);干预72 h PRG组和EPG组仍显著低于PPP组(
P
<
0.05)
且PRG组显著低于EPG组(
P
<
0.05)。干预48 h时
PRG组和EPG组HaCaT细胞内
S. aureus
计数较PPP组显著降低(
P
<
0.05); PRP组与PPP组间差异无统计学意义(
P
>
0.05)。
S. aureus
感染导致HaCaT细胞增殖显著受损(
P
<
0.05)
EPG干预后24 h内未见细胞增殖下降(
P
>
0.05)
36 h后可见细胞增殖显著增加(
P
<
0.05)。
结论
2
本实验建立的模型为糖尿病足溃疡的体外研究提供了一种便利工具
EPG作为PRG的有效成分能够保护受
S. aureus
感染的HaCaT细胞并促进细胞增殖。
Objective
2
To investigate the antibacterial effect of platelet-rich plasma (PRP)
platelet-rich gel (PRG) and extract liquid of platelet-rich gel (EPG) to
Staphylococcus aureus
(
S. aureus
)
and explore the cellular protective effect of EPG on the proliferation of human keratinocytes (HaCaT cells) in an
in vitro
model for infected wounds under high glucose condition.
Methods
2
Fresh whole blood samples were collected from the patients with diabetic foot receiving PRG therapy
and then platelet-poor plasma (PPP) and PRP were harvested by centrifugation. HaC
aT cells and
S. aureus
were co-cultured under high glucose condition to serve as an
in vitro
model for infected wounds. With PPP intervention to the model as control group
the experimental group was divided into PRP intervention group
PRG intervention group and EPG intervention group (
n
=3). Viable bacteria in each group at different time points were counted by plate colony counting. CCK8 assay was applied to detect the proliferation of HaCaT cells in cell culture group
cell and bacteria co-culture group and EPG intervention group.
Results
2
The extracellular
S. aureus
counts in the PRP group
RPG group and EPG group were significantly lower than that of PPP group after 12 and 24 hours' intervention (
P
<
0.05). In 48 h after intervention
the counts in the PRG group and EPG group were obviously lower than that of PPP group (
P
<
0.05)
but there was no notable difference in the count between the PRP group and the PPP group
and between the PRG group and the EPG group (
P
>
0.05). In 72 h after intervention
the counts in the PRG group and the EPG group were also lower than that of the PPP group (
P
<
0.05)
so was in the PRG group than the EPG group (
P
<
0.05).
S. aureus
infection significantly impaired HaCaT cell proliferation (
P
<
0.05). EPG intervention for 12 h resulted in no obvious change in proliferation decline (
P
>
0.05)
and the intervention for 36 h significantly increased cell proliferation (
P
<
0.05).
Conclusion
2
The model for infected wounds under high glucose condition
in vitro
is a reliable
simple and useful tool for diabetic foot. EPG as an effective component of PRG is able to protect HaCaT cells from bacterial damage and promote cell proliferation.
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