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healing between acute wounds, chronic wounds and wounds in the aged
A uthor(s)
Contents
Stuart Enoch
MBBS, MR CSEd, MR C S (Eng)
C linica l R e se a rch Fe llow (PhD)
Patricia Price
BA(Hons), P hD, AFBPsS, C HP sychol
Dire ctor
W ound He aling R e se a rch Unit
Me dice ntre , Unive rsity of W a le s C olle ge of
Me dicine , C ardiff, UK
Em ail: price pe @whru.co.uk
Introduction
Acute (norm a l) wound he a ling
C hronic wound e nvironm e nt and
im pa ire d he a ling
Age ing: e ffe cts on wound he a ling
C onclusion
R e fe re nce s
Key Points
1. Chronic wound healing does not follow the same pattern as that of
acute wounds.
2. A significant number of chronic or non-healing wounds are more
prevalent in older people so the effects of ageing on the healing
process need to be understood and taken into account when assessing
older people.
3. There is an urgent need to develop meaningful alternative endpoints,
as opposed to complete wound closure, for use when assessing
patients with chronic or non-healing wounds.
Abstract
Acute wound healing is a dynamic process involving the coordinated actions of both
resident and migratory cell populations within the extracellular matrix environment
leading to the repair of injured tissues. In contrast to this some wounds fail to heal in
a timely and orderly manner, resulting in chronic non-healing wounds. In addition,
chronic wounds are more prevalent in older people due to the altered molecular and
cellular characteristics of the aged skin and various associated co-morbidities. This
article discusses in detail the cellular, molecular and biochemical differences in healing
between acute and chronic wounds, and outlines the effects of ageing on the healing
process.
Introduction
Skin, the largest organ in the human body, plays a crucial role in the sustenance of
life through the regulation of water and electrolyte balance, thermoregulation, and by
acting as a barrier to external noxious agents including micro-organisms. When this
barrier is disrupted due to any cause - ulcers, burns, neoplasm or trauma - these
functions are no longer adequately performed. It is therefore vital to restore its
integrity as soon as possible.
A wound is defined as a break in the epithelial integrity of the skin. However, the
disruption could be deeper, extending to the dermis, subcutaneous fat, fascia, muscle
or even the bone. Normal wound healing involves a complex and dynamic but superbly
orchestrated series of events leading to the repair of injured tissues. A completely
healed wound, usually seen after simple injury, is defined as one that has returned to
its normal anatomical structure, function and appearance within a reasonable period of
time. It is also defined as one that has attained complete skin closure without
drainage or dressing requirements. In contrast to these some wounds fail to heal in a
timely and orderly manner, resulting in chronic, non-healing wounds. Despite advances
in molecular biology, the development of various tissue-engineered skin substitutes
and growth factors, and a range of other therapeutic options, chronic ulceration
remains a significant problem in our society.
Chronic wounds result from various causes, including venous (chronic venous leg
ulcers or CVLUs), arterial, neuropathic, pressure, vasculitis and burns. Although
chronic ulceration can affect any anatomical region, the most common site is the
lower limb and the estimated prevalence of active leg ulceration in Europe is at least
0.1-0.3 percent [1], [2], [3]. Ulcers secondary to venous hypertension and venous
insufficiency accounts for nearly 70 per cent of all leg ulcers [4], with diabetes and
arterial disease contributing towards a significant proportion of the rest. As all these
conditions are more prevalent in older people they are more susceptible to leg ulcers.
In addition, the increased occurrence and longevity of these ulcers are further
compounded by the detrimental effects ageing has on the skin and the wound healing
process.
This article outlines the cellular, molecular and biochemical differences between the
acute and the chronic wound environment. The effects of ageing on healing are also
reviewed as a significant number of non-healing chronic wounds are prevalent in older
people.
inflammatory response. The alpha granules of the platelets contain growth factors,
including platelet-derived growth factor (PDGF), insulin-like growth factor-1 (IGF-1),
epidermal growth factor (EGF), and transforming growth factor-beta (TGF-). These
proteins initiate the wound healing cascade by attracting and activating fibroblasts,
endothelial cells and macrophages. The platelets also contain dense bodies that store
vasoactive amines such as serotonin that increase microvascular permeability. This
leads to the exudation of fluid into the extravascular space and results in tissue
oedema, although this feature is more prominent during the inflammatory phase.
Early inflammatory phase
The next phase of healing is inflammation, which begins with the activation of
complement and the initiation of the classical molecular cascade that leads to
infiltration of the wound with granulocytes or polymorphonuclear leucocytes (PMNLs).
These cells are attracted to the wound site within 24 to 48 hours of injury by a
number of agents, including complement components such as C5a, platelets, formylmethionyl peptide products from bacteria and TGF-.
Within a short time, the PMNLs begin to adhere to the endothelial cells in the adjacent
blood vessels through a process called margination and start to move through the
vessel wall, a process known as diapedesis. Once in the wound environment they
phagocytose bacteria and other foreign particles, killing them by releasing degrading
enzymes and oxygen-derived free radical species. PMNL activity usually ceases within
a few days of wounding once contaminating bacteria have been cleared. Redundant
cells are cleared away from the wound by extrusion to the wound surface as slough or
phagocytosis by macrophages. The main function of PMNLs is to prevent infection so
they contribute little to the normal wound healing process beyond this stage.
Late inflammatory phase
Blood monocytes undergo a phenotypic change on arrival at the wound site to become
tissue macrophages. Monocytes are attracted to the wound by a variety of
chemoattractants, including complement, clotting components, immunoglobulin G (IgG)
fragments, collagen and elastin breakdown products, and cytokines such as
leukotriene B4, platelet factor IV, PDGF and TGF-. Macrophages are the most
important cells present in the later stages of the inflammatory process (48-72 hours)
and appear to act as the key regulatory cells for repair. They release further
cytokines and growth factors into the wound, recruiting fibroblasts, keratinocytes and
endothelial cells to repair the damaged blood vessels [8]. Macrophages are also
capable of releasing proteolytic enzymes such as collagenase that can debride tissue.
The depletion of circulating monocytes and tissue macrophages causes severe
alterations in wound healing, leading to poor wound debridement, delayed fibroblast
proliferation, inadequate angiogenesis and poor fibrosis. Additional growth factors such
as transforming growth factor-alpha (TGF-), heparin-binding epidermal growth factor
(HB-EGF), and basic fibroblast growth factor (bFGF) are secreted by the PMNLs and
macrophages, which further stimulate the inflammatory response.
The lymphocyte is the last cell type to enter the wound during the inflammatory phase
(>72 hours after wounding) and may be attracted by interleukin-1 (IL-1), IgG and
complement products. IL-1 is believed to play a key role in the regulation of
collagenase, indicating that the lymphocyte may be involved in collagen and
extracellular matrix (ECM) remodelling. The role of lymphocytes in wound healing,
however, has not been clearly defined.
Proliferative phase
The proliferative phase starts at about day three and lasts for two weeks after
wounding. It is characterised by the replacement of the provisional fibrin/fibronectin
matrix with newly formed granulation tissue.
Fibroblast migration: Fibroblasts and myofibroblasts appear in the wound between 2
and 4 days after wounding. Following injury they are stimulated to migrate into the
wound defect, proliferate and produce the matrix proteins fibronectin, hyaluronan (HA)
and later collagen and proteoglycans. Fibroblasts are attracted by a number of factors
including PDGF and TGF- [9]. Once within the wound environment, fibroblasts
proliferate and start to construct the new ECM, which is essential for the repair
process and supports further ingrowth of cells. Interactions between the fibroblasts
and the ECM itself help to determine the synthesis and remodelling of the matrix [10].
Collagen synthesis: Collagens, which are synthesised by fibroblasts, provide
strength and integrity for all tissues in the body and therefore play a particularly vital
role in wound repair. Collagens are a key component of all phases of wound healing.
Immediately after injury, exposed collagen comes into contact with blood, promoting
platelet aggregation and activating chemotactic factors involved in the response to
injury. Later collagen becomes the foundation of the wound ECM. Invading fibroblasts
synthesise and secrete types I and III collagen to form the new matrix.
Angiogenesis: The process of forming new blood vessels occurs concurrently during
all stages of the healing process. TGF- and PDGF, secreted by the platelets during
the haemostatic phase, attract macrophages and granulocytes and promote
angiogenesis. The macrophages, in particular, play a key role in angiogenesis by
releasing a number of other angiogenic substances including tumour necrosis factor-;
and bFGF. Angiogenic capillary sprouts invade the fibrin/fibronectin-rich wound clot
and organise into a microvascular network throughout the granulation tissue within a
few days [11]. As collagen accumulates in the granulation tissue to produce scar
tissue the density of blood vessels diminishes. Disturbance of this dynamic process
may influence the development of chronic wounds [12].
Granulation tissue formation: Granulation tissue is so called because of the pink
granular appearance of numerous capillaries that invade the wound stroma. Each
'granule' contains a loop of capillaries and therefore bleeds easily if traumatised.
Granulation tissue is made up mainly of proliferating fibroblasts, capillaries and tissue
macrophages in a matrix of collagen, glycosaminoglycans (GAGs) including HA, and the
glycoproteins fibronectin and tenascin [13], [14]. Granulation tissue formation is
evident as early as 48 hours after wounding and by 96 hours fibroblasts become the
predominant cell type in this tissue [15].
Epithelialisation: A single layer of epidermal cells start to migrate from the wound
edges within a few hours of wounding to form a delicate covering over the raw area
exposed by the loss of epidermis, a process known as epiboly. From about 12 hours
after wounding there is a marked increase in mitotic activity in the basal cells from the
wound edges or around skin appendages. These cells loosen their normally firm
attachments to the underlying dermis, allowing them to migrate in a leap-frog fashion
across the provisional matrix [16]. When advancing epithelial cells meet, further
movement is halted by contact inhibition and a new basement membrane regenerates.
Further epithelial cell growth and differentiation re-establishes the stratified
epithelium. The rate of epithelial coverage is increased if the wound does not require
debridement, if the basal lamina is intact and if the wound is kept moist. A dry eschar
(scab) slows the rate of epithelialisation. Several growth factors modulate
epithelialisation: EGF is a potent stimulator of epithelial mitogenesis and chemotaxis,
while other growth factors, such as bFGF and keratinocyte growth factor, also
stimulate epithelial proliferation.
Remodelling phase
Matrix synthesis and the remodelling phase are initiated concurrently with the
development of granulation tissue and continue over prolonged periods of time. As the
matrix matures, fibronectin and HA are broken down and collagen bundles increase in
diameter, corresponding with increasing wound tensile strength [17], [18]. However,
these collagen fibres never regain the original strength of normal unwounded skin and
only a maximum of 80 percent unwounded skin strength can be achieved [19].
There is ongoing collagen synthesis and breakdown as the ECM is continually
remodelled, equilibrating to a steady state about 21 days after wounding. Collagen
degradation is achieved by specific matrix matalloproteinases (MMPs) that are
produced by many cells at the wound site, including fibroblasts, granulocytes and
macrophages. As remodelling of the wound continues, MMP activity decreases and
tissue inhibitors of metalloproteinases (TIMPs) activity increases. TGF- plays an
important role in mediating this, underlining the ability of TGF- to promote matrix
accumulation.
Early collagen deposition is highly disorganised but its subsequent organisation is
primarily achieved by wound contraction. Wound remodelling occurs when the
underlying contractile connective tissue shrinks in size to bring the wound margins
closer together. Contraction occurs through the interactions between fibroblasts and
the surrounding ECM. These interactions may be influenced by a number of
extracellular factors including TGF-, PDGF and FGF [10]. With time the density of
macrophages and fibroblasts is reduced by apoptosis triggered by unknown sources
[20]. However, it has been suggested that apoptosis may be signalled by the
withdrawal of cytokines as the wound heals, although many other theories exist
including myofibroblast differentiation itself signalling apoptosis and the release of
certain factors following re-epithelialisation [21]. With continued remodelling the
outgrowth of capillaries is halted, blood flow to the area is reduced and metabolic
activity in the area declines. An acellular, avascular scar is the final result of an acute
wound healing process.
Decreased
Tissue inhibitor of
m e talloprote ina se s (TIMPs)
2 - m a croglobulin
Se rine prote ase s - Urok inase -type plasm inoge n activator, C athe psin
G, incre ase d ne utrophil e lastase a ctivity
Vitrone ctin
Te na scin
Dia be te s m e llitus, Spina l injurie s, C e re bral Pa lsy, Ha nse n's dise ase (Le prosy)
ISC HEMIA
Athe roscle rosis, calcifica tion, m icroa ngiopa thy (dia be te s m e llitus), any form of
pe riphe ra l va scula r dise a se
P ER IP HERAL
O EDEMA
Ve nous hype rte nsion (de e p ve nous throm bosis, va ricose ve ins), syste m ic cause s
(re na l or ca rdiac failure ), lym phoe de m a , de cre ase d album in, e le phantia sis (com m on
in tropical countrie s)
P R ESSUR E
O THER
C AUSES
C onne ctive tissue disorde rs le ading to vasculitis, a rte rio-ve nous m alform a tions, drugs
such as corticoste roids a nd hydrox yure a, syste m ic dise ase s, m alignancy,
oste om ye litis, sm ok ing, inhe rite d ne utrophil disorde rs, poor nutritiona l status
Complications
P re se nce of ne crotic a nd
unhe althy tissue
Ex ce ss e x uda te and slough
La ck of a de quate blood
supply
Abse nce of he althy
granula tion tissue
Syste m ic a m yloidosis
C linica l or sub-clinical
infe ction
There is an up-regulation of MMP-2 in normal aged skin, and MMP-2 and MMP-9 in
acute wounds in aged skin in comparison with young adults [71]. This alteration in the
cytokine profile is similar to that seen in chronic wounds in younger patients. TIMP-1
and TIMP-2 mRNA levels are also significantly reduced in normal aged skin and after
acute wounding in aged skin [72]. These factors may combine to predispose the
remodelled wound to recurrent breakdown. In animal models, a delayed inflammatory
response has been observed after acute wounding in middle-aged and aged compared
to young mice [66]. Similarly, in aged mice a decline in wound macrophage phagocytic
function and a delay in T-lymphocyte response in the later stages of healing has been
observed [73]. Senescence is also associated with the over-expression of IL-1a in
endothelial cells, which inhibits angiogenesis [74] and reduces the expression of IL-6
[75].
In addition to co-morbidities, the clinical impairment of wound healing in the aged may
be related to delayed cellular proliferation, changes in ECM production and
composition, and an altered cytokine and inflammatory response. These factors, on
their own or in combination, contribute to the increased occurrence and longevity of
wounds in older people.
Conclusion
It is clear that the environment and healing patterns of acute and chronic wounds,
including those in older people, are dissimilar. Because of these differences complete
wound closure may not be a realistic outcome in many chronic wounds and wounds in
older people. For this reason the use of meaningful alternative endpoints in the
assessment of chronic, recalcitrant wounds, such as a reduction in exudate, better
pain control or an improvement in overall quality of life, are currently the subject of
intense debate.
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otherwise stated.
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