不同形式富血小板血浆在体外高糖感染创面模型中的抗菌作用

何敏; 李涛; 王敏; 任芮; 陈兵; 邓武权

doi:10.16016/j.1000-5404.201809003

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不同形式富血小板血浆在体外高糖感染创面模型中的抗菌作用

基础医学 | 更新时间：2026-01-09

- 不同形式富血小板血浆在体外高糖感染创面模型中的抗菌作用
- Antibacterial role of platelet-rich plasma of different forms in in vitro model for high-glucose infected wounds
- 第三军医大学学报 2018年40卷第23期页码：2168-2172
- 作者机构：
  
  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
- 作者简介：
  
  [ 邓武权, E-mail: wuquandeng@gmail.com DENG Wuquan, E-mail:wuquandeng@gmail.com
- 基金信息：
  
  国家自然科学基金青年科学基金(81500596);重庆市基础研究与前沿探索专项(CSTC2018jcyjA2721)
- DOI：10.16016/j.1000-5404.201809003
  中图分类号： R457.14;R587.1;R632
- 收稿：2018-05-13，
  
  修回：2018-09-10，
  
  纸质出版：2018-12-15
- 稿件说明：
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何敏, 李涛, 王敏, 等. 不同形式富血小板血浆在体外高糖感染创面模型中的抗菌作用[J]. 第三军医大学学报, 2018,40(23):2168-2172.

Min HE, Tao LI, Min WANG, et al. Antibacterial role of platelet-rich plasma of different forms in in vitro model for high-glucose infected wounds[J]. Journal of Third Military Medical University, 2018, 40(23): 2168-2172.
何敏, 李涛, 王敏, 等. 不同形式富血小板血浆在体外高糖感染创面模型中的抗菌作用[J]. 第三军医大学学报, 2018,40(23):2168-2172. DOI： 10.16016/j.1000-5404.201809003.

Min HE, Tao LI, Min WANG, et al. Antibacterial role of platelet-rich plasma of different forms in in vitro model for high-glucose infected wounds[J]. Journal of Third Military Medical University, 2018, 40(23): 2168-2172. DOI： 10.16016/j.1000-5404.201809003.

摘要

目的

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

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(

=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

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(

0.05);干预48 h PRG组和EPG组显著低于PPP组(

0.05)

而PRP组和PPP组

以及PRG组和EPG组差异均无统计学意义(

0.05);干预72 h PRG组和EPG组仍显著低于PPP组(

0.05)

且PRG组显著低于EPG组(

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(

0.05); PRP组与PPP组间差异无统计学意义(

0.05)。

S. aureus

感染导致HaCaT细胞增殖显著受损(

0.05)

EPG干预后24 h内未见细胞增殖下降(

0.05)

36 h后可见细胞增殖显著增加(

0.05)。

结论

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。

Abstract

Objective

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

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 (

=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

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 (

0.05). In 48 h after intervention

the counts in the PRG group and EPG group were obviously lower than that of PPP group (

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 (

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 (

0.05)

so was in the PRG group than the EPG group (

0.05).

S. aureus

infection significantly impaired HaCaT cell proliferation (

0.05). EPG intervention for 12 h resulted in no obvious change in proliferation decline (

0.05)

and the intervention for 36 h significantly increased cell proliferation (

0.05).

Conclusion

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.

关键词

Keywords

references

ARMSTRONG D G, BOULTON A J M, BUS S A. Diabetic foot ulcers and their recurrence[J]. N Engl J Med, 2017, 376(24): 2367-2375. DOI: 10.1056/NEJMra1615439.

SPICHLER A, HURWITZ B L, ARMSTRONG D G, et al. Microbiology of diabetic foot infections: from louis pasteur to 'crime scene investigation'[J]. BMC Med, 2015, 13: 2. DOI:10.1186/s12916-014-0232-0.

UCKAY I, GARIANI K, DUBOIS-FERRIERE V, et al. Diabetic foot infections: recent literature and cornerstones of management[J]. Curr Opin Infect Dis, 2016, 29(2): 145-52. DOI: 10.1097/QCO.0000000000000243.

PICARD F, HERSANT B, BOSC R, et al. The growing evidence for the use of platelet-rich plasma on diabetic chronic wounds: a review and a proposal for a new standard care[J]. Wound Repair Regen, 2015, 23(5): 638-43. DOI: 10.1111/wrr.12317.

DENG W, BOEY J, CHEN B, et al. Platelet-rich plasma, bilayered acellular matrix grafting and negative pressure wound therapy in diabetic foot infection[J]. J Wound Care, 2016, 25(7): 393-7. DOI: 10.12968/jowc.2016.25.7.393.

MERCIER R C, DIETZ R M, MAZZOLA J L, et al. Beneficial influence of platelets on antibiotic efficacy in an in vitro model of staphylococcus aureus-induced endocarditis[J ] . Antimicrob Agents Chemother, 2004, 48(7): 2551-7. DOI: 10.1128/AAC.48.7.2551-2557.2004.

BIELECKI T M, GAZDZIK T S, ARENDT J, et al. Antibacterial effect of autologous platelet gel enriched with growth factors and other active substances: an in vitro study[J ] . J Bone Joint Surg Br, 2007, 89(3): 417-20. DOI: 10.1302/0301-620X.89B3.18491.

WIEGAND C, ABEL M, RUTH P, et al. HaCaT keratinocytes in co-culture with staphylococcus aureus can be protected from bacterial damage by polihexanide[J]. Wound Repair Regen, 2009, 17(5):730-8. DOI: 10.1111/j.1524-475X.2009.00536.x.

BOWLER P G, DUERDEN B I, ARMSTRONG D G. Wound microbiology and associated approaches to wound management[J]. Clin Microbiol Rev, 2001, 14(2): 244-69. DOI: 10.1128/CMR.14.2.244-269.2001.

HOSOYA A, LEE J M, ChO S W, et al. Morphological evidence of basal keratinocyte migration during the re-epithelialization process[J]. Histochem Cell Biol, 2008, 130(6): 1165-75. DOI: 10.1007/s00418-008-0499-3.

NESTLE F O, DI MEGLIO P, Qin J Z, et al. Skin immune sentinels in health and disease[J]. Nat Rev Immunol, 2009, 9(10): 679-91. DOI: 10.1038/nri2622.

DEPPERMANN C, KUBES P. Platelets and infection[J]. Semin Immunol, 2016, 28(6): 536-545. DOI: 10.1016/j.smim. 2016.10.005.

GIRALDO C E, ALVAREZ M E, CARMONA J U. Influence of calcium salts and bovine thrombin on growth factor release from equine platelet-rich gel supernatants[J]. Vet Comp Orthop Traumatol, 2017, 30(1): 1-7. DOI: 10.3415/VCOT-16-02-0026.

DOHAN D M, CHOUKROUN J, DISS A, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. part Ⅱ: platelet-related biologic features[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2006, 101(3): e45-50. DOI: 10.1016/j.tripleo.2005.07.009.

VÁZQUEZ-SÁNCHEZ E A, RODRÍGUEZ-ROMERO M, SÁNCHEZ-TORRES L E, et al.Peptidoglycan from staphylococcus aureus has an anti-apoptotic effect in HaCaT keratinocytes mediated by the production of the cellular inhibitor of apoptosis protein-2[J]. Microbiol Immunol, 2014, 58(2):87-95. DOI: 10.1111/1348-0421.12126.

CHEN L, WANG C, LIU H, et al. Antibacterial effect ofautologous platelet-rich gel derived from subjects with diabetic dermal ulcers in vitro [J ] . J Diabetes Res, 2013, 2013: 269527. DOI: 10.1155/2013/269527.

BURNOUF T, CHOU M L, WU Y W, et al. Antimicrobial activity of platelet (PLT)-poor plasma, PLT-rich plasma, PLT gel, and solvent/detergent-treated PLT lysate biomaterials against wound bacteria[J]. Transfusion, 2013, 53(1): 138-46. DOI: 10.1111/j.1537-2995.2012.03668.x.

ETULAIN J. Platelets in wound healing and regenerative medicine[J]. Platelets, 2018, 29(6):556-568. DOI: 10.1080/ 09537104.2018.1430357.

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