Microbiology Aspect of Wound Infection:

in Vitro Test for Hydrophobic Interaction

of Hydrophobic Dressing and Efficacy of
Hydrophobic Dressing Compared with
Conventional Dressing

Yeva Rosana, Conny R. Tjampakasari, Beti E. Dewi

Microbiology Department,
Medical Faculty,University of Indonesia
Wound infections

one of the most important and

potentially serious complications
that occur in the acute period
following injury
Anatomy of SKIn
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The wound surface is sterile
immediately following injury
 First 48 hours : repopulated quickly with
gram-positive organisms from hair follicles,
skin appendages, and the environment

 After 5-7 days : more virulent gram-negative

organisms replace the gram-positive
organisms
Common Wound Pathogens
Staphylococcus aureus
- naturally colonises the skin and the upper
respiratory tract
- most frequent wound bacterium with
pathogenic potential occurence of
multiresistant strains: MRSA

Streptococci
- occur naturally in the oral cavity
- typical wound pathogens are Streptococcus
pyogenes and haemolytic Streptococci
(groups A or B)
Common Wound Pathogens
Pseudomonas aeruginosa
- can form water-soluble green pigments that may lead
to green colouring of the infected wound
- often found in burn wounds

Escherichia coli (E. coli)

- occurs in the intestine

Enterococci
- belong to the lactic acid bacteria
 Common Wound Pathogens

Candida albicans
- can be found on mucosa and in the
digestive tract
- can lead to dermal fungal infections

- Others …….
Mechanism of Microbial Infection

Production & release of toxins and

enzymes that affect cell growth Elimination
Adherence of of bacteria
wound patho- and toxins
gens to the
Invasion,
tissue Activation of the
replication,
immune response
spreading

Microbes bind to - Microbes spread in the wound. - By activation of the

- Released enzymes and toxins can body´s immune
specific surface response
structures of damage the body´s own tissue.
- Cell production is inhibited by - Microbe and toxins
the host cells. are eliminated.
microbes toxins.
Bioburden Levels - Definitions
Contamination
 Every wound (even the so-called aseptic surgical wound) is
+/- populated by bacteria e.g. deriving from the skin flora
 There is low microbial replication (=reproduction) in the wound or
soft tissue
 Hence pathogens can be inactivated by the immune system

Colonisation
 Bacterial replication takes place but without affecting the wound
healing process, remains in check by immune system
!
Critical Colonisation
 The point at which bacterial presence has a negative impact on the
wound healing process
!!!  No visible infection signs, often increased exudation and pain

Infection
 Microbial replication resulting in a high microbial load
 The wound healing is impaired or stopped completely
 The immune response is activated, classical infection signs occur
(redness, swelling, hyperthermia, pain, functional restrictions)
 Local infection / spreading infection (sepsis)
 Types of Wound Infection

◦ Pyogenic infection: production of pus

 Staphylococci
 Streptococci
 Pseudomonas aeruginosa
 Escherichia coli
Putrid infection: sullage-smelling tissue
 Escherichia coli
 Proteus vulgaris
Gas gangrene: swelling of the tissue due to toxic effects caused by
Clostridia
 Clostridium perfringens
 Clostridium novyi
 Clostridium septicum

Tetanus
 Clostridium tetani
 Management of Wound Infection

1.Burn Wound Infection Continuum

Antimicrobial therapy
2.Wound Exudate Continuum
Absorptive products
Antimicrobial Therapy
 Treatment with antibiotics
(local, systemic)

 Treatment with antiseptic agents

(such as iodine or silver)

 Treatment without using chemically active

agents (by physical interaction)
 Antibiotics

 Chemical compounds that inhibit or abolish the growth of

microorganisms

Narrow-spectrum antibiotics target particular types of

bacteria, e.g. only Gram-negative
. or Gram-positive species
Broad-spectrum antibiotics target a wide range of
species

 Their efficacy depends on the location of the infection, the

ability of the antibiotic to reach the infection site, and the
ability of the microbe to inactivate or excrete the antibiotic
 Antibiotics
Systemic antibiotics

 Usually use of oral antibiotics

 Intravenous antibiotics only in serious cases, e.g.
systemic infections .
 At serious wound infection or spreading infection,
treatment by oral antibiotics should be considered
in addition to the local antimicrobial wound
treatment
 Bacterial Antibiotic Resistance
Development of resistance
 Potential survival of bacteria at encounter with an antibiotic e.g.
methicillin
 The surviving bacterium can mutate (alteration of the genetic
information) and might develop resistance to the antibiotic substance
 Replication of the antibiotic-resistant bacterium and potential spreading
 Increasing number of antibiotic-resistant bacteria strains

Causes
 When patients do not complete the full course of antibiotic therapy
 Overuse of antibiotics allows bacteria to develop resistance to a wide
range of antibiotics
Antiseptics
 Chemical substances that prevent the growth or activity of
microorganisms
◦ Bactericidal action: bacteria are destroyed
◦ Bacteriostatic action: bacterial growth/replication is inhibited

 Applied topically to living tissue

◦ As contrary to disinfectants that are used in non-living material such
. as surfaces or instruments

 In wound care, various antiseptic agents (e.g. wound

cleansing solutions, iodine) or antiseptic dressings (e.g. silver
dressings) are used for local antimicrobial therapy
• Hydrophobic dressing :

A promising method using a

purely physical method without
chemically active agents to
reduce morbidity and mortality
rates from wound infection
 Hydrophobic properties those are
expressed by microbial cell surface
structures are essential for
microbial life
 Microorganisms need hydrophobic
surface structures to provide:
 Cell to cell commmunication (e.g. DNA
exchange)
 Binding to surfaces for nutrition
 Protection against phagocytosis
 Adherence to host tissue (e.g. as initial
phase of wound infection)
 Examples of hydrophobic tissue
adhesions include:

 Fimbriae of Gram-negative bacteria

 Cell surface proteins of fungi
 S-layer proteins (capsule-like
polysaccharide surface coatings)
 Lipoteichoic acid of Gram-positive bacteria
 Since most pathogenic microorganisms that
impair wound healing have hydrophobic
properties
 using the hydrophobic dressing on wound
area will reduce the microbial load and
activate or support the natural wound healing
process.
 The microorganisms multiply to quite a low
extent when absorbed in the dressing, and
may not produce extracellular toxins and
enzyme.
What is hydrophobic interaction?
A basic physical principle:
Two hydrophobic (water repellent) particles bind to each other
with help from the surrounding water molecules that form a circle
around the particles - like a jacket - and hold them together.
Hydrophobicity of microbes

Fact:
Wound pathogenic bacteria and
fungi express cell surface
hydrophobicity

The more virulent, the more hydrophobic

Binds and
removes
microbes
from colonised
and infected
wounds
•MATERIALS AND METHODS
• This was a cross section study that was done in the
Department of Microbiology, Faculty of Medicine
University of Indonesia, on January 2009

• Methicillin-resistant Staphylococcus aureus

• Pseudomonas aeruginosa
 cultured on nutrient agar and harvested after 18 to
24 hours of incubation.
 The bacterial concentration was adjusted to 0.5
MacFarland (1.5 x 108 CFU/ml) by Nephlometer
equipment,
• immediately used for the binding assays.
METHODS:
Binding assay

Step 1 Step 3
wound dressing Step 2 washed three times
materialswere cut incubated with 1 ml of
into 1 cm2 sheets for 5 minutes each with
bacterial
under sterile 0.07 M PBS, pH 7.2, and
solution at various placed in nutrient broth
condition incubation time
METHODS:

Step 4 Step 5
The nutrient broth were The number of bound
incubated at 35-37oC for 18 to 24 bacteria were determined by
hours Nephlometer equipment
Summary
 Wound infection is a serious problem
because it causes a delay in epidermal
maturation and leads to additional scar
tissue formation.
 Invasion of microorganisms into the tissue
layers below the dermis may also result in
bacteremia, sepsis, and multiple-organ
dysfunction syndrome.
Summary
 A hydrophobic dressing is a unique mode
of action to bind and inactivate bacteria
and fungi in area of wound infection by
hydrophobic interaction without using any
antiseptic or antibiotic agents.
 Using the hydrophobic dressing on wound
area will reduce the microbial load and
will activate or support the natural
wound healing process.