Rachel Simmons, Molly Gibson, Breda Cullen, Claire Bartle, Lorna McInroy, Tanya Swaine Systagenix, Gargrave, UK

1. EP508
EWMA May 2013
Effect on Bacterial Protease Activity
Rachel Simmons, Molly Gibson, Breda Cullen, Claire Bartle, Lorna McInroy, Tanya Swaine
Systagenix, Gargrave, UK
Effect of Collagen/ORC/Silver‐ORC
on Bacterial Proteases
 Bacterial proteases, virulence factors
associated with active pathogenesis and
expressed during infection have been
suggested to be detrimental to the wound
healing process.
 This study evaluated if a
Collagen/ORC/Silver‐ORC matrix* could aid
in bacterial load reduction and bacterial
protease activity reduction in chronic
wounds.
Assay for Bacterial Protease Activity
Effect on Bacterial Protease Activity Clinically
Log10 Reduction
Conclusions
C1385‐ 3
*PROMOGRAN PRISMA®, Systagenix

2. Effect of Collagen/ORC/Silver‐ORC on Bacterial Protease Activity
en/ORC/Silver‐ORC* was tested against a range of commercially available bacterial
ases as well as Staphylococcus aureus and Pseudomonas aeruginosa supernatants to
mine its ability to reduce the activity of bacterial proteases in vitro.
V8 from S. aureus Pseudomonas Protease from B. Bacterial protease Thermolysin form Protease from S.
Commercially Available Bacterial Proteases
013

3. Effect of Collagen/ORC/Silver‐ORC on Bacterial Protease Activity
(continued)
en/ORC/Silver‐ORC reduced the protease activity of supernatants and commercially
ble bacterial proteases. While reduction was varied for different bacterial proteases
majority of species less than 10% activity remained after 24 hours.
0
20
40
60
80
100
Bacterial Supernatants
Neat Protease
24 hour incubation with
Collagen/ORC/Silver‐ORC
013

4. Assay for Bacterial Protease Activity
terial protease activity was measured using a casein florescent activity assay;
strate commercially available
fer: 25mM Tris, 150mM NaCl, pH 7.2
asure at Ex 485nm/ Em 518nm
d fluid was collected from a patient with a 4 month mixed venous/arterial chronic
measuring 2cm x 1cm. Bacterial proteases activity was measured throughout the
on of treatment to determine how Collagen/ORC/Silver‐ORC matrix affects levels of
rial proteases in a wound environment.
Effect of Collagen/ORC/Silver‐ORC on Bacterial Protease Activity Clinically
013

5. ‐5
0
5
10
15
20
0 1 2 3 4
Relative Units Activity/ min
Weeks
Bacterial Protease Human Neutrophil elastase Total MMP
Initiation of Collagen/ORC/Silver‐ORC matrix
se of Collagen/ORC/Silver‐ORC matrix reduced the bacterial protease activity
Effect of Collagen/ORC/Silver‐ORC
on Bacterial Protease Activity Clinically (continued)
013

6. Log10 Reduction
n/ORC/Silver‐ORC was tested against 10 bacterial strains, including Methicillin‐
t Staphylococcus aureus (MRSA), Methicillin‐Resistant Staphylococcus epidermidis
and Vancomycin‐resistant Enterococci (VRE), on Log10 reduction assay to determine
microbial properties of this matrix.
en/ORC/Silver‐ORC showed efficacy on the Log10 reduction assay against all strains of
a tested, with reduction to min. detectable limits after 24 hrs
013

7. Log10 Reduction
obacter
mannii
19606
Enterococcus
faecalis ATCC
49532
Enterococcus
faecalis (VRE)
PHLS 33
Enterococcus
faecium (VRE)
PHLS 26
MRSA PHLS 2 MRSE NCTC
11964
Proteus
mirabilis NCTC
9559
Pseudomonas
aeruginosa
ATCC 27312
Staphylococcus
aureus ATCC
6538
Steptococcus
pyogenes NCTC
10871
Mean Log10 reduction values for Collagen/ORC/Silver‐ORC against a range of bacteria over 24
hours
013

8. Conclusions
agen/ORC/Silver‐ORC is effective at reducing the bacterial load of common
hogens associated with chronic wound infection and reducing the activity of
terial proteases both in vitro and in vivo.
shows that Collagen/ORC/Silver‐ORC is effective at protecting against infection by
only preventing bacterial growth but also by modulating the pathogenic / virulent
vity of bacterial proteases, which have been suggested to be detrimental to the
und healing process.
013