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 ›




‡ 

 
(insulation & evaporation)
‡ 

  
 
 Detects touch,
pressure, pain and temperature
‡ Protects underlying tissues and organs by acting
as 
 
 
against desiccation,
invasion by microbes, environmental insults
such as UV irradiation, mechanical trauma,
chemical or thermal burns
‡ Excretes salts, water and organic wastes
(through the sweat glands)
‡ Synthesis vitamin D3
‡ Dermis ± no regeneration
‡ Repair by fibrosis
‡ Pig skin = human skin ± model
‡ Most studies ± rodents or rabbit ±
breeding, handling and maintenance
‡ Wound healing studies ± difficult to
compare ± differences in model, strain,
sex, age, location and size of wound
à
 
 à

Structure of the Adult Mammalian skin
‡ Two main layers ±epidermis & dermis
‡ Y
Y

±  
  

 
± consisting primarily of 


 from various
stages of differentiation, from mitotically active basal
cells (stratum basale-deepest layer) to the heavily
keratinized superficial cells (stratum corneum) -
sloughed off
± Keratinocytes ± impermeable sheet ± tight junctions,
desmosomes
± 


  
- constantly divide to
self renew and give rise to differentiated
keratinocytes that migrates upwards to replace the
cells of the stratum corneum as they slough off
YY ›Y
Y

 EPIDERMIS

Non epithelial cell

types :



-
colour





APCs




mechanoreceptors
‡ The ECM of the stratum basale contains
hyaluronic acid (HA) for which the basal cells
express the CD44 receptor.
 Y

‡ located beneath the epidermis
‡ two layers of fibroblasts which are embedded in
ECM ;
‡ 
  
- next to the basal layer of the
epidermis ± papillae
‡ Deeper 
  

‡ contains most of the skins¶ specialized cells and
structures, including: Blood & Lymph vessels, Hair
follicles, Sweat & Sebaceous glands, Nerve
endings, Collagen & Elastin
 The Papillary Layer
‡ thrown into folds of highly vascularized




!


‡ pervaded by a  

" -
nourishment to the epidermis, and acts as
a heat exchanger
‡ ECM ± thin collagen and elastic fibers;
mast cells; tissue macrophages; fat cells
 The Reticular Layer
‡ thicker than the papillary layer
‡ characterized by an ECM containing a network
of  
collagen & elastin fibers
‡ larger blood vessels and fewer capillaries
‡ The reticular layer rests on a  

  
 (not part of the skin)
‡ Bundles of collagen fibers extending from
reticular layer anchor it onto the hypodermis
 



Y
‡ three classes ± Proteoglycans, Fibrous Proteins,
Adhesive Proteins
  Y #$
‡ proteins linked to sulfated
GAGs
‡ Dermatan sulfate, heparan
sulfate & chondroitin sulfate -
prominent in dermal ECM
‡ Significant PGs in dermal
ECM are the large PG
versican, and the small PG
decorin
‡ Multiple PGs linked to HA
‡ Binds water - - resisting
compressive forces and space
for cell migration in injured skin
 FIBROUS PROTEINS
‡ Major Fibrous Proteins in the Dermal ECM
‡ 

- give tensile strength
± type I (80%) and type III ± major
± Type VI - forms a highly branched network of
filaments surrounding the type I collage fibrils
± type IV - part of basement membrane of blood
vessels
± type VIII - located around hair follicles, and
small blood vessels
‡ Y
 give resiliency - allowing the skin to be
stretched and then assume its original shape
 The Dermal Adhesive Proteins
‡ Prominent - Fibronectin (Fn), Vitronectin (Vn),
and Tenascin-C (Tn-C)
‡ › 



% 


serve as a
substrate to which cells can adhere when they
are either migrating or stationary.
‡ 


 is an antiadhesive protein - with
Fibronectin, helps control the degree of cellular
adhesion to the ECM substrate



 

‡ Proteins of dermal ECM ± aa recognition
sequences ± receptor binding
‡ & 


  Y
 
 such
as collagen I, III, and Fibronectin
‡ low affinity, heterodimeric linker proteins
consisting of two non covalently associated
transmembrane glycoprotein subunits ' 
(
‡ Functions:
± adhesion of cells to ECM
± migration of cells on ECM
± maintenance and modulation of gene expression by
the transmission of signals to the nucleus
‡ Epidermal Growth Factor Receptor- Tenascin-C
and type I collagen have EGF repeat domains -
bind to EGFR
‡ Other recognition domains ± protein binding ±
organization of ECM
‡ Basement membrane ± synthesized by
epidermis ± connects with papillary layer
± Composed of laminin in lamina lucida
± Type IV collagen, entactin and perlecan ± lamina
densa
± Epidermal cells ± lamina lucida ± hemidesmosomes
and integrins
± Lamina densa- papillary layer ± type VII collagen
‡ The dermal ECM - reservoir for GFs - binding &
releasing them upon injury
‡ GFs are signaling molecules that stimulate or
inhibit proliferation, migration and differentiation,
depending upon the cell type
‡ Most GFs - ) " - initiate intracellular
phosphorylation cascades by other kinases,
resulting in activation of transcription factors that
up-or down-regulate gene activity
‡ TGF-beta ± Smad pathway
‡ ECM, GFs, CAM and GF receptors ± imp for
wound healing
The Effect of Wound Type & Extent on
Dermal Repair
‡ Injury to the epidermis layer alone - regeneration
without scar formation
‡ wounds that penetrate the dermis - repair by
fibrosis
‡ Wounds made by  " 


* 
+ - simple fibroblast proliferation with
minimal scar formation - little wound space to be
filled in
‡ 




, excisional wounds and burns -

wound edges - far apart ± destruction of
substantial amounts of tissue - repair by the
formation of extensive scar tissue
Phases of Repair in Excisional Wounds
‡ 3 tightly integrated phases - occupy variable time
frames, depending on size of wound:
 Hemostasis
 Inflammation
 Structural Repair
‡ Nine cell types - major roles in the epidermal and
dermal repair process:
‡ 

,

  ,  
,
,

,
&
-


 






!
 - hemostasis and inflammation
‡ Y

,
 





 - means for structural repair
 

.
/
› 

‡ Wound - blood vessels, epidermal and
connective tissue cells & surrounding ECM
‡ First response to wounding ± HEMOSTASIS -
minutes to stop bleeding and seal off the wound
by 3 mechanisms:
a) Formation of Platelet Clumps
b) Vasoconstriction
c) Formation of Fibrin Clot
 +› 
 

 
Immediately upon an injury,  into the
wounded area

The cells in the wall of blood vessel at the site of injury -



 -  


 to the injury site


 

of clumped platelets upon contact with
exposed collagen in the walls of the injured blood vessels

releasing more ADP (further attracts more platelets),

Arachidonic acid, Fibrinogen, Fibronectin,
Thrombospondin, and von Willebrand Factor VIII

AA - converted to thromboxane A2 through COX pathway

 +%
 

‡  -

0



from injured
nerve axons ± !
   - restrict blood
flow into the wound by constricting the blood
vessels
‡ The injured nerves -  

 , a
neuropeptide - 

 

in the
dermis - releasing 

± increase in the
permeability of vessel walls -allowing further
leakage of plasma into the wound
 +› 
› 

‡ Blood plasma in the wound space contains  

  - induce clot formation
‡ 


› %

, a plasma protein initiates a
cascade of reactions involving about 10
  (I-
XII) and require 02 as an essential cofactor.
‡ 
 - produced at the end of cascade and
converts prothrombin to active enzyme  

‡ Thrombin catalyzes the conversion of plasma  



 
, the major structural protein of clot
Prothrombin Thrombin
Tissue factor

Fibrinogen Fibrin
(insoluble clot)
‡ Fibrin molecules -e organized into fibers that
intertwine to form a 
 "  - traps RBCs,
WBCs and platelets
‡ Meshwork also contains plasma › 


,
% 


as well as  

from
degranulated platelets and 



,



,
%*probably synthesized by monocytes)

Blood Clot Formation (blood cells, platelets, fibrin clot) (SEM x10,980)

‡ Contraction of provisional matrix and
exudation of serum ±  



‡ Dehydration of clot ± scab ± prevents fluid
loss
‡ Invasion by cells of immune system
 
03



Within a day after injury, 
  released during
clot formation







 of surrounding capillaries - attracts
neutrophils, monocytes and T-lymphocytes - crawl out
between endothelial cells into the provisional fibrin matrix

The leukocytes insert themselves into spaces between

endothelial cells via binding of Platelet Endothelial Cell Adhesion
Molecule * Y+ on the surface of the leukocytes to PECAM
on extrusions of endothelial cell surface.

$
  


 migrate into the clot
simultaneously; neutrophils in greater numbers - use fibronectin
in the clot as an adhesive substrate and express the appropriate
integrin receptor to bind to fibronectin
 After entering the wound and adhering to Fibronectin, the







  


Within the clot, macrophages secrete TGF-ȕ, PDGF and other

chemoattractants such as leukotriene B4, monocyte-
chemoattractant proteins 1,2,3 (MCPs), macrophage
inflammatory protein 1Į and ȕ, and the chemotactic protein
CAP37 ± attract ± neutrophils and macrophages

Neutrophil influx diminishes within 3-4 days after injury. T-cell

lymphocyte peaks by the end of the first week or later. T cells
particularly the Th1 subset may play a role in regulating
macrophage-induced activities.
 Phase 2: Inflammation

‡ Lasts for 5-7d

‡ Within a day - 
  - released
during platelet degranulation and clot formation
including  


(fibrinogen cleavage
by thrombin) ,  

 
  
produced by the action of 
,



 

,

&


fragments
‡ The most important chemoattractants for the
inflammatory phase are #›( and #›
supplied by degranulating platelets.
‡ #›( - attracts monocytes
‡ #› attracts neutrophils and monocytes
‡   ± plasminogen to plasmin
‡ 
± activates   and also acts
directly on fibrin
 Important tasks of inflammatory
neutrophils and macrophages
‡ kill bacteria through oxygen-dependent mechanisms that
generate 0 0 
 
‡ Neutrophils also produce 


 and
proteins, including the enzyme 

#
‡ Neutrophils and macrophages also play a 

  



 


within the wound - production of
  - MMP-1 and MMP-8
‡ MMP-1 degrades type I and III collagens & MMP-8
degrades type I collagen.
‡ The neutrophils and macrophages 

 
"


by phagocytizing dead bacteria, as well as cellular
and molecular debris.
‡ a

   
  

  

 
 



‡ 








 
 
 

 



 






  


 


‡ 
 
 


 

 

 
 
 
‡ a

  
  
 






 
 

 
 Phase 3 : Structural Repair

‡ Several sub phases:




4

› 
 # 






  # 



 
 Phase 3: Structural Repair
a) Re-epithelialization
Within a few hours after injury - 

  


of the epidermis at the edge of wound 


 
 

 to one another and begin  



 through the fibrin clot to cover the wound surface

The migrating cell sheet just expands (does not divide) -

basal cells just interior to the wound edge divide and
feed progeny into the migrating sheet

After the migrating epidermis has expanded to cover the

wound surface, its 
!
!
 





 and synthesize a new basement membrane
Excessive damage ± no regeneration of hair and sweat glands
 b) Development of Granulation Tissue




 
    

Within two days after injury, well before the inflammatory

phase is over,  

 
 from the
surrounding dermis into the clot.

Simultaneously , new capillaries regenerate into the

nascent fibroblastic tissue, which is now called
 


because the capillaries give it a
reddish, granular appearance

Initiated by GFs secreted by macrophages, primarily

#›
#›(  stimulate fibroblast migration and
proliferation
Macrophages, fibroblast and capillaries ± invade the wound
space as a
± biologically interdependent during tissue
repair

The fibroblasts synthesize a new transitional matrix to

replace the fibrin matrix, and the new capillaries carry
oxygen and nutrients to the injury site, to sustain the
metabolism required for these cellular process.
›  

 


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acts as a
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wound
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‡ Cytokines:
PDGF ± produced by macrophages

± IL-1 (Interlukin) ± by macrophages ±

stimulates fibroblasts to produce PDGF

± IFN-ȕ ± inhibits fibroblast entry into G1

± TGF- ȕ & TNF-Į (Tumor necrosis factor) ±

stimulate fibroblast proliferation

‡ (


 

fibroblasts invading the wound
space increases 


 




   

fibrin provisional =atrix is
degraded 


 




  

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 plas=inogen activator
inhibitor(pAI) 

 

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conversion of plas=inogen to plas=in"
 YsM Synthesis By Fibroblasts
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signs of fibrotic
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‡ peripheral sensory and sy=pathetic
postganglionic nerves - regenerate 
 




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role in the for=ation of
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neuropeptides such as substance p   


 

 

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‡ Final phase of structural repair - 




   


  
acellular
fibrous scar tissue
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± shondroitin-4-sulfate p is =uch higher

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 MMps '  ' 


lysyl oxidase 
 


 





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der=al contraction
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unwounded skin
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 nor=al pattern of collagen
synthesis and architecture
‡ Fetal wound fibroblasts ʹ synthesize higher
levels of HA and HA receptor    


  
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‡ Sulfated p synthesis 

 

 
 

 


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 higher type III/type I collagen
ratio
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infla==atory response
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receptors 


 


 
 



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‡ ound environ=ent ʹ =ajor deter=ining
factor
‡ Yxtent of injury ' r