2003

BIOTECHNOLOGY
CONVENTION
 Artificial Skin
Technology
Integra LifeScience Corporation:
Ms. Murad, Dr. Amirrtha

Genes R Us:
Dr. Jacqueline, Dr. Naghmeh

M.C.: Vuk Pavlovic

 Skin
Skin is the largest organ in the body
 Facts
Every year in the United States, 1.1 million
burn injuries demand medical attention
Ten thousand people die every year of
burn-related infections
 Solution
INTEGRA Dermal Regeneration Template
Originally conceived and designed by Dr. John
F. Burke and Dr. Ioannas Yannas
INTEGRA . . .
provides immediate wound closure
is an innovative bilayer matrix that provides a scaffold
for dermal regenration
is a biodegradable template that induces organized
regeneration of dermal tissue by the body
 And now for the biotechnology . . .

Source: Design of an Artificial Skin. I, II, III, Journal of Biomedical

Materials Research, Dagalakis et al.
Stages of Clinical Experience
Stage 1: Physiochemical and Mechanical
constraints
Keep bacteria out
Prevent tissue and organ water from
evaporating
Stage 2: Biochemical constraints
Minimize formation of deep, disfiguring scars
and contractures
 Preventing Infection
Keep bacteria out
Damaged Skin Graft

Air Bubbles

Wound Bed
 Air Pocket Prevention
Wetting the woundbed
Surgically remove dead tissue
Bending rigidity of membrane must be small
High peel strength
accidental shear force should not pull
membrane off
 Moderate Fluid Flux /
Moisture Flux Rate
Must be maintained
Too high graft dehydrates and air
pockets form
Too low interface compromised
Rate should be close to human
physiological rate
 Artificial Membrane
No need to mimic all properties of skin
Membrane only needs to have:
Large tear strength
Appropriate Elasticity
Typical thickness should be 0.1 to 0.5 mm
< 0.1 mm Difficult to suture
> 0.5 mm Not flexible enough to wet
woundbed effectively
How is Even Growth and Healing
Ensured?
Biodegradability of the membrane
Pores in the membrane
How is Even Growth and Healing
Ensured?
Biodegradability of the membrane
Must biodegrade in 25 days
Faster degradation membrane is reduced
to a liquid-like state
Slower degradation membrane must be
surgically removed
How is Even Growth and Healing
Ensured?
Pores in the membrane
Cell migration must occur pores must be >
10 m
Migration
Depends on availability of gases and nutrients
depends on diffusion
The dermal layer can regenerate within 25
days
The epidermal layer cannot a temporary
synthetic is used until epithelial cells can be
cultured
Selection of Chemical Compounds
For dermal layer a cross-linked
collagen-glycosaminoglycan coprecipitate
is used
For epidermal layer Silastic is used
Membrane Compound Criteria
Polypeptides and polysaccharides
can degrade to non-toxic compounds
structurally similar to macromolecules in the
woundbed
can exist in highly hydrated forms
 Why Collagen?
Hydrophilic polymer
Biodegradability (which can be controlled
by cross-linking)
Weak antigen low rejection
Tough enough for surgical applications
Physical properties are well understood
leads to design flexibility
 Why a Glycosaminoglycan
(GAG) Coprecipitate?
Coprecipitate is more flexible than cross-
linked collagen
Biodegradation can still be controlled
Significantly more porous
Weak antigen
Degrades into non-toxic compounds
Drawback to GAG precipitate
GAG can be eluted out
Cross-linking can prevent this
 Why Silastic?
Silastic is a synthetic medical grade
flexible silicone sheet
Strong
Flexible
Controls water flux like real skin
 Preparation of Collagen
From bovine hide
Collagen is depolymerized by liming
Sliced to thin pieces
Washed with acid for 4 hours
Water removed through filter
Pulverized to tiny pieces
Freeze-dried yielding loose, shredded
mats
 Preparation of GAG
From shark cartilage
GAG used as received from Sigma
Chemical company
 Preparation of Membrane
Freeze-dried collagen flakes
pulverized to minute particles
dispersed in acetic acid

ACID (A)
 Preparation of Membrane
Freeze-dried collagen flakes
pulverized to minute particles
dispersed in acetic acid
GAG dispersed in acetic acid
GAG added drop-wise to
collagen while stirred

ACID (B)
 Preparation of Membrane
GAG

Freeze-dried collagen flakes

pulverized to minute particles
dispersed in acetic acid
GAG dispersed in acetic acid
GAG added drop-wise to
collagen while stirred
Dispersion stirred for Collagen

additional 5 minutes
 Preparation of Membrane
Precipitate is then dehydrated by one of
3 different methods
1. Casting solvent evaporation at room
temperature
2. Filtration in Buechner funnel fitted with
filter (assisted by aspiration)
3. Direct freeze-drying at 60 oC
The various methods affect pore size
 Preparation of Membrane
The formed membrane is then cross-
linked
Cast films and filtered membranes are
placed in glutaraldehyde bath
Excess aldehyde is removed
Freeze-dried membranes are cross-
linked differently
 Preparation of Membrane
Cross-linking of Freeze-Dried Membrane

First, the membrane is treated in a

vacuum oven so it doesnt collapse in an
aqueous solution
Then, the membrane is bathed in
glutaraldehyde
Cross-linked membrane is freeze-dried
and stored
 Collagen-GAG Membrane
Formation Summary
Collagen Dispersion
GAG Solution

Coprecipitation + Homogenization

Casting, Filtration or Freeze-Drying

Cross Linking

Freeze Drying and Storage

 Integra
This membrane is the dermal layer of artificial
skin
The epidermal layer is made of Silastic
The two fused together make Integra

Silastic

Integra

Membrane
Biological Properties of Artificial
Skin
Facilitate rapid migration of fibroblasts and
microvascular cells (neodermis synthesis)
Absent inflammatory and foreign body reaction
Synthesis of normal neodermal tissue
Controlled rate of biodegradation
Low/no antigenicity
Nontoxic metabolites
Prevent scar formation and contracture
Infection prevention
Mechanical Properties of Artificial
Skin
Epidermal portion impermeable to bacteria
Moderate fluid flux
Tear strength
Bending rigidity
Control of pore structure in dermal portion
Modulus of elasticity
High peel strength
Stable handling and suturing
characteristics
 The Healing Process

Source: Integra LifeSciences Corporation

The Healing Process
The Healing Process
 Advantages of Artificial Skin
Wound Closure
Availability in large quantities
Long shelf life
Durability
Permeability
 Advantages of Artificial Skin
Good Adherence
Bacterial defense
Ease of application and removal
Cosmetic Acceptability
Feels more like natural skin than
autografts or cultured human cells.
Advantages of Artificial Skin
Advantages of Artificial Skin
 Complications and Unusual
Cellular Responses
Loss of Integra due to a haematoma under graft
Wrinkling of Integra over the arm or lower leg
Sometimes associated with collection of sterile fluid
under the Integra
Silastic sometimes loosens at edges 2 to 3
weeks after placement
In areas of wear or motion caused by joint movement
Infection can occur in the graft bed under Integra
Fewer lymphocytes formed than expected
during the healing process
Complications and Unusual
Cellular Responses
 The Future of Integra
The dermal matrix of Integra seeded with
autologous human keratinocytes

In vitro seeded
Integra
(scanning
electron
microscopy)
 Artificial Skin
Technology
Questions and Answers . . .