Tissue repair:

cell regeneration and

fibrosis
Dr Maha Akkawi
 Tissue repair
 Repair involves the combination of two
processes, regeneration and fibrosis.
 Regeneration: healing of the injured tissue by
regrowth of the parenchymal cells of the same
type.
 Fibrosis: healing of the injured tissue by
connective tissue resulting in scar formation.
 Tissue repair
 In any tissue healing, both processes are
involved but in different proportions
depending on:
 Type of tissue injured.
 The preservation of the underlying tissues
framework.
 The severity of the injury.
 Tissue repair
 Both processes involve essentially similar
mechanisms including
 Cell migration, proliferation and differentiation.
 Synthesis of and interaction with the extracellular
matrix, ECM.

However, the cell types involved are different.

 Cell regeneration

 Definition:
Parenchymal cell proliferation and
differentiation, which involves interaction of
the proliferating cells with soluble chemical
mediators and insoluble extracellular matrix.
 Cell proliferation

 Cell cycle: A series of checkpoints and

defined phases through which proliferating
cells progress.
 Phases of the cell cycle
 G0 phase: pool of resting cells
“quiescent cells” are present.
Most cells in the body are
quiescent.
 G1 phase: presynthetic growth
phase.
 S phase: DNA-synthetic phase.
Both of those two phases
constitute the majority of the cell
cycle.
 G2 Phase: premitotic growth
phase.
 M Phase: mitotic phase. This is
the shortest phase.
Types of cells in
relation
to the cell cycle
Proliferative potential of different cell types:
According to the proliferative capacity and relation to
the cell cycle, the different cells in the body are
divided into:
 Labile cells: continuously dividing and dying cells.
Stem cells are the source of this ability. The stem cell
divides to produce one daughter cell retaining the
ability to divide, and one cell that differentiates to
carry out the normal function.
Hematopoietic cells, surface epithelial cells, mucosal
surfaces.
Proliferative potential of different cell types:

 stable cells: they are equivalent to quiescent cells.

They are normally non dividing but are capable of
undergoing rapid division in response to injury.
Parenchymal cells of most solid organs are of this kind as
well as the endothelial cells, the fibroblasts and smooth
muscle cells.
 permanent cells: terminally differentiated
nonproliferating cells in the post natal life.
Neurons and cardiac muscles are typical examples.
Cyclins
 A family of proteins that
control the entry and
progression of the cells
through the cell cycle.
 Cyclins are synthesized
according to the phase, they
change their levels and
activities to control the
phases of the cell cycle and
then they are degraded by
the ubiquitin-proteasome
pathway.
 Cyclin-dependent kinases: CDK
 Constitutively synthesized proteins, which are
activated through complexing with cyclins.
 The cyclin/CDK complex is then
phosphorylated and activated.
Cyclin-dependent kinases: CDK
 Constitutively
synthesized proteins,
which are activated
through complexing
with cyclins.
 The cyclin/CDK
complex is then
phosphorylated and
activated.
 CDK
 The P/cyclin/CDK complex phosphorylates
proteins (kinases phosphorylate proteins)
leading to conformational changes that
 activate or inactivate an enzymatic activity
 induce or interfere with protein-protein interaction

 induce or inhibit binding of a protein to DNA

 induce or prevent the catabolism of a protein.

 CDK inhibitors

 They are important proteins involved in the

regulation of the cyclin/CDK complexes.
 They are especially important at the cell cycle
checkpoints: points at which the cells takes
stock of whether its DNA is sufficiently
replicated and all mistakes repaired before
progressing.
 Proliferative potential of different
cell types:
 This cell growth and differentiation involves at
least two signals: soluble chemical mediators
like growth factors and inhibitors and
insoluble elements of the ECM.
 Soluble mediators:
 Mostly soluble growth factor proteins derived from the
serum or cells.
 Secreted in extremely low concentrations.
 They bind to a specific high-affinity receptors on the
target cells
 Pleiotropic effect: not only stimulate cell growth but
migration, differentiation and remodeling.
 They induce cell proliferation by affecting the expression
of genes involved in normal growth, protooncogenes.
 Some have growth inhibition effects, e.g. TGF-beta.
 What are the general intercellular
signaling pathways for soluble
mediators?
 Gap junctions: narrow hydrophilic channels
effectively connect two cells’ cytoplasm,
permitting the movement of small molecules
between adjacent cells.
 Autocrine: the mediator acts on the cell that
secretes it. Examples are cytokines in immune
response and compensatory hyperplasia .
 What are the general intercellular
signaling pathways for soluble
mediators?
 Paracrine: mediators affect cells only in the
immediate vicinity. Examples include recruitment of
inflammatory cells to the site of infection, and in the
process of wound healing.
 Endocrine: mediators are released into and carried
through the blood stream. Examples include
hormones.
 Synaptic: neurotransmitters are secreted into synapses
at the neuronal-neuronal junctions or the neuronal-
muscle junctions.
What are the general intercellular
signaling pathways for soluble
mediators?
 Types of receptors:

 Intracellular receptors
 Ligand must be hydrophobic to enter the cell e.g. Vit D,
steroids and thyroid hormones.
 Receptor-ligand complex directly associate with nuclear

DNA to activate or tune off gene transcription.

 Types of receptors:

 Cell surface receptors:

 Ligand interacts with surface receptors (4-types).
 Most receptors have an internal enzymatic activity.
 Phosphorylation of substrates with generation of 2ry
messengers like Ca, c-AMP, inositol triphosphate, or the
activation of a kinase, like MAP kinase.
 Amplification of the signal and translocation of activated
transcription factor into the nucleus.
 Conformational changes take place in the DNA, which
enhances or inhibits gene transcription.
 Effect is either stimulation or inhibition of growth.
Cell surface receptors
Types of cell surface receptors:

 Ion-channel receptors: Acetylcholine receptor.

 Receptors with intrinsic kinase activity. PI3 kinase
and MAP kinase. EGFR and FGF.
 G-protein-coupled receptors: Epinephrine and
glucagons and chemokine receptors
 Receptors without intrinsic kinase activity: They
undergo conformational changes that allows
activation of an intracellular protein kinases.
Cytokines and erythropoietin receptors.
 Extracellular matrix; ECM:

 Definition: A macromolecular complex that is

synthesized locally and constitute a large
proportion of any tissue.
 It is dynamic and consistently remodeling
 Forms of ECM

 Interstitial matrix:
 A three dimensional amorphous gel, present in the spaces
between cells in connective tissue, and between epithelium
and supportive vascular and smooth muscle structures
 Synthesized by mesenchymal cells
 Collagens (fibrillary and nonfibrillar), proteoglycan and
glucoproteins are the major constituents.
 Forms of ECM
 Basement membrane:
 A highly organized and specialized matrix, present
around epithelial, endothelial and smooth muscle
cells.
 Synthesized by epithelial and mesenchymal cells

 Type IV collagen and adhesive glucoproteins are

the major constituents.
 Components of the ECM:

 fibrous structural proteins that confer tensile

strength (collagen) and recoil (elastin)
 water-hydrated gels that permit the resilience
and lubrication (proteoglycan and hyaluronan).
 Adhesive glucoproteins and integrins that
connect the matrix elements to one another
and to cells (fibronectin, Laminin).
Components of the ECM:
 Collagen:

 Fibrous structural proteins conferring tensile strength.

 Rope like structure braided into triple helix
 Glycines present at every third position giving them the ability
to tightly intertwine.
 Approximately 18 subtypes are known.
 Fibril collagens form lateral cross linking of the triple helices,
thus giving them the tensile strength. Examples include types
I, III and V.
 Cross linking is dependent on Vit C.
 Nonfibrillar collagens are mainly seen in the B.M. type IV
collagen is the example.
 Elastin:

 give the tissue the ability to recoil and returns back to the
baseline structure after physical stress
 Central core of elastin protein surrounded by a meshwork of
fibrillin glycoprotein.
 Elastin require glycine at every third position, but has fewer
cross-linking
 Fibrillin serves as a scaffold for the deposition of elastin and
the assembly of elastic fibers.
 High content in tissues requiring recoil like walls of large
vessels, uterus, skin and ligaments
 Defects in fibrillin are associated with Marfan syndrome.
 Proteoglycan and hyaluronan:

 Highly hydrated compressible gels conferring resilience and

lubrication.
 Consist of a long polysaccharide called glycosaminoglycans
linked to a protein backbone.
 They provide resilience and lubrication like in cartilage in
joints. They act also as reservoir for growth factors like FGF.
They are also integral cell membrane proteins.
 Examples include dermatan sulfate, heparin sulfate and
syndecan.
 Hyaluronan consists of multiple disaccharide repeats without
protein core. It has the capacity to bind to volumes of water in
the ECM.
 Fibronectin:
 The major component of the interstitial ECM.
 A large disulfide-linked heterodimer.
 Synthesized by fibroblasts, monocytes and
endothelium
 Associated with cell surfaces, BMs, and pericellular
matrix.
 It has specific domains for collagens, fibrin, heparin
and proteoglycan.
 It attaches to cell integrins via a tripeptide arginine-
glycine-aspartic acid “RGD” motif which is
important in cell–ECM adhesion.
Fibronectin
 Intergrins:

 Transmembrane heterodimeric glycoprotein.

 The intracellular domains associate with cytoskeletal
elements
 Mediate the firm adhesion of leukocytes during
migration in inflammation
 Bind to the RGD motifs signaling cell attachment and
affecting locomotion, proliferation, or differentiation.
 Laminin:

 the most abundant glycoprotein in the BM.

 Connects the cells to the ECM like type IV
collagen and heparan sulfate.
 Important in cell survival, proliferation,
differentiation and motility.
Laminin
 Role of ECM:
 mechanical support for cell anchorage.
 Determination of cell orientation (polarity). Basolateral versus
apical are important in most cells
 Control of cell growth
 Maintenance of cell differentiation
 Scaffolding for tissue renewal. Labile and stable cells are
capable of regeneration only if the underlying stroma (B.M) is
maintained.
 Establishment of tissue microenvironments, and so the identity
to cells
 storage and presentation of regulatory molecules. FGF is
stored and released from the B.M. of epithelial cells.