Acknowledgments must be made to Glenda Motta Associates who coordinated the clinical case series.

Mo|ecu|ar m|crob|a| eco|ogy as a d|agnosnc too| to |dennfy mode of acnon and new targets for s||ver wound dress|ngs
L|ndsay ka|an
1*
, M| 2hou
2
, M|che|e Su|tor
3
, kodrecht
1
and8en[am|nW||||ng
2
1LxclLon1echnologleslnc. , LdmonLon, AlberLa, Canada,*lkalan[exclLonLech.com, 780-248-3883, 2ueparLmenL of AgrlculLure, lood and nuLrluon Sclences, unlverslLy of AlberLa, LdmonLon, A8, Canada, 3WesLvlew PealLh CenLer, SLony laln, A8, Canada




olymlcroblal communlues colonlze all wounds and blolms are hypoLheslzed Lo be a
key llnk Lo Lhe chronlc sLaLe and sLalled heallng. 1hese lnfecuons needs Lo be Lackled
by sLudylng mlcrobes and Lhelr ecology uslng molecular meLhods as only a small
fracuon of wound bacLerla are culLured by currenLly used meLhods and sLudles have
shown llule agreemenL beLween culLure and molecular based approaches. Some
lnLervenuons llke dresslng wlLh oxldlzed sllver can re-acuvaLe Lhe sLalled wound onLo
Lhe normal heallng Lra[ecLory buL Lhe underlylng mechanlsms are dlmculL Lo
measure. "#$% %&'() *$+% &, '%- +,.-/'.*0 +-&#,(% &, $(-123) &#- +$/0,4$*.
/,+5,%$2,1 $1 /#0,1$/ 6,'1(% *1( +-*%'0- &#- /#*17-% $1 6,'1( +$/0,4$,&* *1(
4$,4'0(-1 &#*& 50,+,&- #-*.$17 6#-1 %$.8-0 (0-%%$17 /,1&*$1$17 %$.8-0 ,9)%*.&% $%
'%-(:
Introducnon and Cb[ecnves
Samp||ng Schedu|e
I|gure 1: 1he mlcroblal llfesLyle ln a blolm can be llnked Lo phases of mlcroblal conLamlnauon
Lhrough Lo lnfecuon and subsequenL lmpalred heallng ln a wound. lnLervenuon LhaL dlsrupLs
mlcroblal growLh, lncludlng blolmcan allow heallng Lo resume.
Colonization
Contamination
Critical Colonization
Infection
In
t
e
r
v
e
n
n
o
n

Week 1
Week 4
I|gure 2: A) Mlnlmum 8lolm lnhlblLory ConcenLrauon (M8lC) assays
were performed on blolm populauons of I) !"#$%& (!M109), II) '"
)*+,-&.$/) (AC1), and III) 0"),+*,/ (A1CC 23923) grown ln Lhe presence
of varylng concenLrauons of Ag compounds for 4h ln SlmulaLed Wound
lluld (SWl). n > 4 Su. 8) 1he Mlnlmum 8lolm Lradlcauon
ConcenLrauon (M8LC) of 24h esLabllshed blolms grown for 24h ln SWl
and Lhen LreaLed wlLh varylng concenLrauons of Ag compounds for 24h.
n > 4 Su.
Harrison et al. 2008. Nature Rev Micro
Phillips et al. 2010. Wounds International

CuSO 4
AgNO 3
Ag 2SO 4
AgO
Ag 2O
AgOxysalts
0
20
40
60
80
500
1000
1500
2000
Metal Ion
C
o
n
c
. o
f M
e
ta
l [
M
]
MBEC
E.coli
P.aeruginosa
S.aureus
CuSO 4
AgNO 3
Ag 2SO 4
AgO
Ag 2O
Ag Oxysalts
0
10
20
30
40
50
450
850
1250
Metal Ion
C
o
n
c
. o
f M
e
ta
l [
M
]
MBEC
E.coli
P.aeruginosa
S.aureus
A) 8)
1) Ag can exlsL wlLh dlerenL oxldauon sLaLes and aqueous avallablllLy
2) Ag
2+/3+
has anumlcroblal and anublolm acuvlLy aL lower concenLrauons Lhan Ag
1+
&. 1&2+$ (see gure 2).
3) Ag
2+/3+
reduces lnammauon and promoLes wound heallng &. 1&2+$ and ln mouse lnclslonal wounds ln Lhe absence of bacLerla (see gure 3).
rev|ous Work
I|gure 3: unlnfecLed mouse lnclslonal wounds were LreaLed wlLh or wlLhouL
wound dresslngs conLalnlng Ag CxysalLs and assessed aer 3 or 7 days. A) Wound
area measuremenLs and assessmenL of eplLhellallzauon found Ag CxysalLs
slgnlcanLly reduce wound area & promoLe re-eplLhellallsauon. n > 3 Su. 8)
lmmunohlsLochemlsLry of Lhe wounds revealed slgnlcanLly fewer neuLrophlls
and macrophage ln Ag CxysalLs LreaLed wounds compared Lo unLreaLed. n > 3
Su


0.0
0.S
1.0
W
o
u
n
d
A
r
e
a
(
m
m
2
)

=<0.0S
=<0.0S
Contro|
Ag Cxysa|ts
0
100
200
300
400
N
e
u
t
r
o
p
h
|
|
s
(
m
m
2
)

3 Days 3 Days 7 Days 7 Days A) 8)
Lxper|menta| Methods and Study Cr|ter|a
1ab|e 2: anent demograph|c and descr|pnve wound character|sncs
anent 1 2 3 4 S 6 7 8
Age]Sex 78/l 72/l 69/M 60/M 66/M 47/M 70/M 43/M
Under|y|ng d|sease
ACuM,
P1n,uv1,
Morbldly
obese, P1n
luuM, un, vu,
CAu
8urn
1rauma
ACuM,
P1n, vu,
un
CAu, vu ACuM Lu
Wound 1ype vLu SSl Abdomen ulu Skln Lear ulu ulu ulu vLu
Wound Duranon before study
2 mo 6 mo 3 mo 6 wks Several yrs > 6 weeks > 6 weeks 3-8 yrs
kecurr|ng wound yes no unknown no yes yes yes yes
Wound S|ze 1=0 (cm^3) 0.32 8.12 0.24 0.108 0.24 0.27 0.072 0.912
Wound S|ze 1= 4 weeks (cm^3)
0.04 1.16 0.144 0 0.364 0 0 0.264
c|osure 87.S0 8S.71 40.00 100.00 -S1.67 100.00 100.00 71.0S

34567 38,%2 $./*2 8&)9*2*/ :*%%&2,/ ;<=>* ??@A ?4467 ?./,%&.B8*>*.8).2 8&)9*2*/ :*%%&2,/ ;<=>* ?@A
C<D7 C=>*+2*./&$.A 4E<7 4**> 1*&. 2F+$:9$/&/A 4'D7 4&)9*G# >*+&>F*+)% .*,+$>)2F=A 'E47
',%:$.)+= 1)/#,%)+ 8&/*)/*A H347 H$+$.)+= )+2*+&)% 8&/*/)/*A I47 I&1*+ 8&/*)/*
V1
Swab Debr|dement
V2
Week1 Week2 Week3 Week4
Dress|ng Change
V3 V4 VS V6
Day 1 1=0
V1
1ab|e 1: Inc|us|on and Lxc|us|on Cr|ter|a
Inc|us|on Lxc|us|on
Age >17 yrs SysLemlc anubloucs ln prevlous 2 weeks
Chronlc non-heallng wound > 6 weeks known skln sensluvlLy Lo dresslng componenLs
8equlres aL leasL weekly care aruclpauon ln anoLher sLudy
Cutcomes
1ab|e 3: Summary of resu|ts
# genera]swab samp|e 8ange: 9-69, Medlan 29 13
# genera]debr|dement samp|e 8ange: 16-93, Medlan 23 19
1ota| number of genera observed 97
of wounds w|th change |n m|crob|a| compos|non over nme 100
of wounds c|os|ng 87.3
anent 3
T= W4 T= W1
8acter|a| Genera : >10
Plgh proporuon of 02+*>2$#$##,/ sp. on all vlslLs.
lrom ussue swabs 92 relevanL abundance from
1=0 (before Ag oxysalL appllcauon) Lo 41 relauve
abundance on week 4 of LreaLmenL.
Note: DNA ana|ys|s does not |nd|cate ||ve]dead ce||s.
vlslble slgns of redness reduced Wound heallng - spllL lnLo 4 smaller areas
8y Week 4 wound area
reduced 90

CllenL was very pleased
wlLh dresslng
8acter|a| Genera: >3S
At t=0: >80 0" ),+*,/
W4: 8educed Lo 23 relauve abundance ln ussue swabs
Wound fully healed on
nexL follow up vlslL
8acter|a| Genera: >2S
At t=0: MosLly beLaproLeobacLerla (Comamonadaceae)
W4: Malnly 3%%$9)#,%,:sp. and H),%$9)#2*+ sp.
(assoclaLed wlLh skln).
uepLh lmproved (30) by u4 Wound Pealed
Conunued lmprovemenL
8acter|a| Genera : >S0
At t=0: Plgh abundance of 02)>F=%$##$#,/ sp. and
')*.&9)#&%%,/ sp" W3: 02)>F=%$#$##,/ sp. reduced Lo
<2 abundance.
T=0
0
10
20
30
40
30
60
70
80
90
100
S3-1 S3-2 S3-3 S3-6
k
e
|a
n
v
e
A
b
u
n
d
a
n
c
e
(
) p
e
r
s
a
m
p
|e

Allobaculums sp.
epLonlphllus sp.
llnegoldla sp.
SLrepLococcus sp.
SLaphylococcus
peuenkofer
SLaphylococcus sp.
aenlbaclllus sp.
arabacLeroldes sp.
1=0 D1 W1 W4
T=0
anent 7
T= W2 T= W4
0
10
20
30
40
30
60
70
80
90
100
S7-2 S7-3 S7-4 S7-3
k
e
|a
n
v
e
a
b
u
n
d
a
n
c
e
(

)
p
e
r
s
a
m
p
|e

8oseaLeles sp.
CurvlbacLer sp.
Comamonadaceae famlly
8urkholderlales order
Sphlngoblumsp.
CaulobacLeraceae famlly
epLonlphllus sp.
Pelcococcus sp.
llnegoldla sp.
Anaerococcus sp.
S. aureus
SLaphylococcus sp.
Macrococcus sp.
aenlbaclllus sp.
CorynebacLerlumsp.
Acunomyces sp.
1= D1 W1 W2 W3
anent 1
0
10
20
30
40
30
60
70
80
90
100
S1-2 S1-3 S1-6
k
e
|a
n
v
e
a
b
u
n
d
a
n
c
e
(
) p
e
r
s
a
m
p
|e
8oseaLeles sp. CurvlbacLer sp.
lamlly Comamonadaceae lamlly 8urkholderlales
lamlly CaulobacLeraceae lusobacLerlumsp.
Allobaculumsp. 8umlnococcus sp.
SLrepLococcus sp. LacLobaclllus sp.
LacLobaclllus sp. aenlbaclllus sp.
lamlly 8lkenellaceae arabacLeroldes sp.
8acLeroldes sp. roplonlbacLerlumsp.
1=0 1= W2 1= W4
1=0 1= W2 1= W3
anent 8
0
10
20
30
40
30
60
70
80
90
100
S8-1 S8-2 S8-3 S8-4 S8-3 S8-6
k
e
|a
n
v
e
a
b
u
n
d
a
n
c
e
(
) p
e
r
s
a
m
p
|e

lamlly Comamonadaceae
CurvlbacLer sp.
lamlly Comamonadaceae
lamlly 8urkholderlales
Sphlngomonas sp.
lamlly 8hodosplrlllaceae
lamlly 8radyrhlzoblaceae
lamlly CaulobacLeraceae
epLonlphllus sp.
llnegoldla sp.
SLrepLococcus sp.
S. aureus
aenlbaclllus sp.
CorynebacLerlumsp.
1= D1
1=0 D1 W1 W2 W3 W4
D1 W2 W4
Conc|us|ons
1he bacter|a| compos|non of a wound bed |s comp|ex and d|verse
Intervennon w|th a top|ca| s||ver oxysa|t dress|ng changes the bacter|a| compos|non
More data a|ong w|th changes to b|omarkers |n the host response shou|d be co||ected
Acknow|edgments
Lxc|ton 1echno|og|es Inc.: SupporL of Lhe presenLed research
NSLkC: l8ul Lo L.k and Lngage granL Lo 8W
A|berta Innovates 1echno|ogy Iutures: r&u AssoclaLes lellowshlp Lo Lk
Westv|ew nea|th Center: auenL care and sample collecuon
now |s th|s measured |n a chron|c wound?