biofilms hpump

SV Y6: L Thnh Duy

Biofilms in chronic woundsMng Sinh hc trong vt thng mn tnh

HPUMP Surgery Club 2016

Chronic wounds ? ( VT mn tnh?)Khng lnh < 30d (Matthew Malone et al 2016)Khng lnh < 6w ( Collins 2002)

Lm tn thng m ti ch -> cu trc v chc nng khng phc hi y ( Davis 1992)

VT khng lin theo tin trnh ca qu trnh lin thng.


Wound of 5 weeksHPUMP Surgery Club 2016


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What is a biofilm/Mng sinh hc l g?L 1 cu trc chung ca cc t bo vi khun bm dnh vo nhau bn trong lp mng cao phn t ngoi bo v bm dnh vo b mt tr hoc b mt vt sng (Castrilln et al.2010)


Stages of development of biofilms Cc giai on pht trin ca mng sinh hc ( MSH) Nghin cu trn cc Vi khun E.coli, P.aeruginosa, Bacillus subtilis and S.aureus (Lopez et al 2010) v Candida and Aspergillus fumigatus (Kumamoto, 2002, Mller et al 2011) thy c 5 giai on:Bt u VK dng Ph Du ( t do)Bm dnhTng hp cc cao phn t ngoi boHon thinPht tnHPUMP Surgery Club 2016


Ph du vs Biofilm/MSHT do di chuynNhim trng cp tnhp ng vi iu tr n liu phpD nui cyC nhBao bi lp EPS ( cao phn thu ngoi bo)p ng vi mi trngNhim trng mn tnhKhng p ng vi n tr liu phpKhuyn co dng a tr liu phpKh nui cyHPUMP Surgery Club 2016

Clinical significance of bacterial biofilms in chronic wounds Lm sng ca MSH trong VT mn tnhVt thng chm lin (>30d), khng lin. nhim trng c nhng t cp tnh / nn mn tnh Gy ti pht, ghp da -> cht.Cc cht b mt ti to nhanh ( 24-72h)Khng sinh tht bi hoc ti nhim sau dng thuc.p ng tt vi corticosteroidCy vi khun m tnh Why?


Pathogenic biofilm in chronic woundsMSH gy bnh trn VT mn tnh do: Duy tr tnh trng vim cao ti vt thng

Gy ra phn ng vim v kch hot nhiu gen lin quan n MMPs. Cc proterase ny ph hy nhng m lnh v m hi phc -> mn tnh

Gy gim s di chuyn cc t bo Keratin.

EPS hnh thnh 6-12h v trng thnh trong 24h, c c hi trong 24h iu tr v lm gim c planktonic v biofilm.


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Pathogenic biofilm in chronic wounds MSH gy bnh trn VT mn tnh do: khng tt hn vi hng ro min dch, khng sinh tr liu, tia UV, cht dit khun.Chng chia s nhng k nng v kh nng c nhn trong vic tn ti ca nhm/Vt thng mn tnh

Kch hot cc phn ng ha hc.

Heterogeneous, EPS, Biofilm mechanisms, microenvironment.Theo Wound InternationalHPUMP Surgery Club 2016

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Identitication tests and antibiotic susceptibility in biofilms kim tra v xc nh tnh nhy cm khng sinh trong mng sinh hc

Optical microscope Techniques of hybridizationFluorescein staining Molecular probe 16SRNA Specialized staining techniques

(Veeh et al., 2003)



SEM image showing attachment and biofilm formation by Staphylococcus epidermidis.

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Evidence for biofilms in chronic woundsBng chng ca MSH cc VT mn tnh


Why are biofilms so tolerant ?Ti sao cc mng sinh hc li bn vng



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Mechanism of Biofilm ToleranceCn kit khng sinh (the agent does not reach the biofilm)Thm nhp chm (giving biofilm cells a chance to initiate stress responses)p ng stress (cells change activity in response to environmental challenge)Thay i vi mi trng (metabolically inactive, though viable cells)Tip tc tn ti (spore-like cells that may survive an antimicrobial challenge)


C ch p ng stress (3)Thay i v tr mc tiuGim kh nng tip cnTng cng o thiBt hot

Abreu et al. Nat. Prod. Rep., 2012,29


Antibiotic Resistance BiofilmVanc vs Staph biofilm >1000xTobra vs Pseudomonas biofilmMatthew Malone at al 2016


Systemic AntimicrobialsCc khng sinh ton thnCh c tc dng vi VK c hot ng chuyn ha l im bt li nht i vi m ca vt ch do kh nng kch hot gy vim.Phn ln cc mng sinh hc lp su khng hot ng.

Ch 2532% cc khng sinh c tc dng ln mng sinh hcWhy?Moss, A.H., Vasilakis, C., Holley, J.L. et al. Use of a silicone duallumen catheter with a Dacron cuff as a long-term vascular access for hemodialysis patients. Am J Kidney Dis 1990; 16: 3,211-215.

Marr, K.A., Sexton, D.J., Conlon, P.J. et al. Catheter-related bacteremia and outcome of attempted catheter salvage inpatients undergoing hemodialysis. Ann Intern Med 1997; 127: 4, 275-480.HPUMP Surgery Club 2016

Systemic AntimicrobialsCc khng sinh ton thnTiu dit mng sinh hc tm thi

Cc du hiu nhim trng thng ti pht v khng sinh ch tc dng vi cc t bo pht trin nhanh chng b mt.HPUMP Surgery Club 2016



SEM image showing bacteria cocci in cluster (MRSA) 23

Khng sinh tht bi hoc ti nhim khun sau khi dng khng sinhKhng sinh tht bi l du hiu ca nhim trng c mng sinh hc. Vic s dng khng sinh phi hp kim sot mng sinh hc vn ang gy tranh ci. Ngi ta cho rng thiu mt chin lc song song v kim sot mng sinh hc hiu qu ca khng sinh c th ch t 25%-30%Bng 1. Du hiu lm sng ca mng sinh hc trong vt thng mn tnh v bng chng h tr m qu mc / dch titC bng chng v vic m qu mc khin mng sinh hc pht trin(12)M ht ln khng tt (vd: Yu, mc qu mc)Gnh nng sinh hc c th biu hin nh mt m ht yu(13)Du hiu v triu chng nhim trng ti chDu hiu nhim trng th pht thng l kiu nhim trng c mng sinh hc (14)Cy vt thng m tnh Cy thng thng s ch bt c vi khun dng tri t do (ph du) ch khng bt c dng mng sinh hc (17, 18)Vt thng khng lnh mc d qun l vt thng ti u v h tr c thNhim trng ko di trn 30 ngyNhim trng di 30 ngy cng vn c th c mng sinh hc, nhim trng vi khun t do khng tn ti qu 30 ngyMng sinh hc khng vi: Tia cc tm, cht dit khun, khng sinh, h thng min dch. Mng sinh hc c th ti cu trc rt nhanh nhng v nguyn tc th khng tiu dit vt ch.p ng vi corticosteroid v c ch TNF - alphaVim l mt sn phm ph ca mng sinh hc, do , p ng tt vi iu tr vim cho thy s hin din ca mng sinh hc. Gim vim loi b ngun dinh dng chnh. (15)D dng loi b khi b mt vt thng GelatinCc nh lm sng v nghin cu ang c gng xc nh liu cc sn phm ph ca vic hnh thnh mng sinh hc c c pht hin trn lm sng. Cc ca nghin cu ch ra rng cc cht trong vt thng c th b loi b d dng song cng ti to rt nhanh, c trong vt thng v di bng gc. Mt vi tc gi cho rng cc hoi t t tng t nh mng sinh hc, nhng kt lun cha c chng minh. Kt hp gia cao phn t t tit v mt s thnh phn ca vt ch gi n mng sinh hc (20,21)Cc cht b mt ti to rt nhanhNghin cu ch ra rng mng sinh hc c th ti to trong vng 24-72 gi (22)HPUMP Surgery Club 2016

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Treatment of Biofilms... HPUMP Surgery Club 2016

Ti sao kh iu tr mng sinh hc?

Do tng cng cc protein h tr chng li khng sinh.im mnh trong tnh khng ng nht S hp tc gia cc loi, biu hin cc protein khc nhauVi khun nm su trong mng sinh hc t hot ng trao i cht v n nu trnh cc mi e da.Mng sinh hc c th ti to rt nhanh trong 24hHPUMP Surgery Club 2016

How to treat biofilm bacteria?iu tr MSH nh th no?

Nn iu tr a tr liu.


Lm schLoi b c hc v trit .Vi khun phi tr li trng thi hot ng trao i cht, EPS cha hon thin, trng thi gn kt yu > vi khun nhy cm hnTng nhy cm vi khng sinh trong vng 24-72 gi u sau lm sch.HPUMP Surgery Club 2016


Topical AntimicrobialsCc Khng sinh ti chSilverCadexomer Iodosorb

Bng chng cho thy Bc c hiu qu ph v biofilm in-vitro1.Chaw et al, Antomicrob Agents Chemo, 20052.Bjarnsholt et al, APMIS, 20073.Percival et al (2008) 4.Kostenko et al. Antimicrobial Agents and Chemotherapy. 2010.

C kh nng ngn chn v ph v mng sinh hc trng thnh ca trc khun m xanh (P. aerguinosa in-vitro):

1.Phillips, P. L. et al., Effects of Antimicrobial Agents on an (in-vitro) ADVANCES IN WOUND CARE (2010).HPUMP Surgery Club 2016

Tm tt v chm sc vt thng c mng sinh hcLm sch thng xuyn v tch cc

Ra nn vt thng s dng khng sinh b mt

Khng sinh ti ch bc , cadexomer iodosorb

Khng sinh ng ung mnh v lu di

Hiu qu ca p lc m ti ch trn vt thng c mng sinh hcThng xuyn quan tm, chm sc bnh nhn ca bn.HPUMP Surgery Club 2016

VAC: Vacuum Assisted Closure


Ci thin s khuch tn oxy n t bo,Bin i vi mi trng hng thun li cho s lnh vt thngTng cng phng thch yu t tng trng, sn xut cht nn, co vt thng v tng sinh t bo (hnh thnh m ht v biu m ha)

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CASE


Khm bnh 2 ngy/ln

Hng dn BN ra b mt vt thng hng ngy

Hng dn dng Cadexomer Iodosorb sau ra sch

Loi b c hc nhng m cht bng gc or ct lc

iu tr KS theo kinh nghim Ciprofloxacin 7daysHPUMP Surgery Club 2016

S lng (copies by qPCR) D61M16090HPUMP Surgery Club 2016

Day 7HPUMP Surgery Club 2016

S lng (copies by qPCR) D62M1042

Gim s lng vi khun n 94%HPUMP Surgery Club 2016

BIOFILMS


Thank you so muchHPUMP Surgery Club 2016

Author/sStudy designNumber of participantsVisualisation methods with or without molecularChronic wound aetiologiesNo of samples with confirmed biofilm (%)

James et al (2008)Prospective studycase vs control

66Light microscopy, SEM16S rRNA with DGGE13 DFUs, 21 PUs8 VLUs, 24 NHSW30 out of 50(60%)

Kirketerp-Moller (2008)Prospective cohort study22PNA-FISH, CLSM

Un-specified chronic wounds13 of 22(60%)

Fazli et al (2009)Prospective cohort study

9PNA-FISH, CLSM10 VLUs10 of 10(100%)

Thomsen et al (2009)Prospective cohort studySub analysis2PNA-FISH, 16S rRNA2 VLUs2 of 2(100%)

Han et al (2011)Prospective cohort study

15PNA-FISH, CLSM16S rRNA4 DFUs, 5 PUs, 2 VLUs4 NHSW9 of 15(60%)

Neut et al (2011)Case report2CLSM2 DFUs2 of 2(100%)

Oates et al (2014)Prospective cohort studySub analysis4FISH, SEM,

4 DFUs4 of 4(100%)

Martinez-Velasco et al (2014)Prospective cohort studyconference abstract

20SEM, LMUn-specified chronic wounds20 of 20(100%)

Honorato-Sampaio et al (2014)Prospective cohort study45TEM45 VLUs45 of 45(100%)

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