Lecture title - Repair and regeneration

70 important questions on Lecture title - Repair and regeneration

What are the three tissue injury outcomes

1. Resolution
2. Regeneration
3. Repair

What is resolution (tissue injury outcome)?

Removal of inflammatory exudate
No tissue damage = no damage to original architecture of tissue

What is regeneration (tissue injury outcome)?

Proliferation of same cell type as lost cells
Reconstruction of original architecture
May leave no residual trace of injury
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What is repair (tissue injury outcome)?

Replacement of lost tissue by connective/fibrous tissue instead of original cell
May lead to scar
Complex tissue architecture may not be reconstructed

What is needed to resolve damage to the epithelium but not the underlying tissue?

Regeneration

What is needed to resolve a more severe injury that involves damage to the epithelium and the connective tissue?

Scar formation

Which stage of the cell cycle are stable/quiescent cells in when they are not proliferating?

Stage G0 of the cell cycle

When is the restoration of normal tissue architecture possible?

Only when the residual tissue is structurally intact

What type of damage can cause scarring from restoring the normal tissue architecture?

Damage from infection/inflammation can cause scarring

How are injured cells of the epithelia restored?

Injured cells of the epithelia are replaced by proliferation of residual cells (neighbouring cells of the injury) and differentiation of stem cells.

What determines whether regeneration of the epithelia succeeds?

1. Extent of damage
2. Integrity of stem cells
3. Underlying support, like the basement membrane, must be intact
(basement membrane guides epithelial regeneration)

What is the regeneration of parenchymal tissue with stable cells like?

It is limited other than the liver

Why does repair by scarring happen?

1. Extensive tissue damage
2. Damage to permanent/non-dividing cells

What are the two possible scenarios of repair if the lost tissue can't be replaced?

1. Replacement of injured cells with connective tissue -> scar
2. Regeneration of some residual cells + scar formation

List the overall steps of scar formation with durations of each step

1. Inflammation (6-48h) = removal of dead tissue and microbes by phagocytosis
2. Cell proliferation (up to 10 days) = covering of wound by epithelial cells, angiogenesis by endothelial and vascular cells, production of collagen fibres that later form scar = granulation tissue production
3. Remodelling (2-3 weeks - months) = reorganisation of connective tissue deposited by fibroblasts for fibrous scar formation

What is the purpose of remodelling for scar formation?

Breaking down of a layer of original tissue ensures that new collagen fibres can be strongly laid down to the original tissue

What is a granulation tissue?

New connective tissue that has formed on the surface of a wound during healing with pink soft granular appearance.

What are the histological features of granulation tissue?

1. New small capillary loops = bright red stain
2. Fibroblasts and myofibroblasts = spindle-shaped cells
3. New and mature collagen = blue stain

Compare the histological characteristics of granulation tissue and mature scar

Granulation tissues = numerous blood vessels, oedema, loose extracellular matrix, occasional inflammatory cells 
Mature scar = dense collagen, scattered vascular channels  

What induces angiogenesis?

Vascular endothelial growth factors (VEGF)

Where do fibroblasts enter a wound from during repair?

From the wound edges, migrating toward the centre

What cell type can some fibroblasts differentiate into during healing?

Myofibroblasts

What protein is expressed by myofibroblasts?

Smooth muscle actin, which increases contractile activity

What is the purpose of increased contractile activity during healing by myofibroblasts?

To contract and close the wound by pulling margins of the wound to the centre

What is the purpose of wound contraction?

To reduce the volume of tissue that needs to repaired by around 80%

Explain how wound contraction works

1. Myofibroblasts attach to each other and adjacent matrix
2. Granulation tissue as a whole contracts to draw the surrounding tissue together

What are the possible problems that can occur due to wound contraction?

Stricture, contracture, improper wound healing after burns to skin

What are the problems of wound contraction after burns to skin?

Cosmetic damage, reduced motility

Define extracellular matrix (ECM)

Complex of proteins that assemble into a network surrounding cells and takes up a large portion of any tissue

What are the major functions of the extracellular matrix (ECM)?

1. Mechanical support - cell anchorage, cell migration
2. Control of cell proliferation - growth factors, signalling
3. Scaffold (temporary physical framework) for tissue renewal

What are the general components of the extracellular matrix (ECM)?

1. Fibrous structural elements - collagen, elastin
2. Gels - proteoglycans, hyaluronan
3. Adhesive glycoproteins / link proteins - fibronectin, laminin

What do the fibrous structural elements of the extracellular matrix (ECM) do?

Provide strength and recoil

What do the gels of the extracellular matrix do?

Provide resilience and lubrication

What do the adhesive glycoproteins and link proteins of the extracellular matrix (ECM) do?

Connect matrix elements and cells

How does the role of the fibroblasts and new vessels change as repair progresses?

Fibroblasts and newly formed vessels decrease in number, remaining fibroblasts adapt a more synthetic phenotype, producing the extracellular matrix

When is collagen synthesized during wound healing?

In the early stages of wound healing, from 3-5days to weeks, depending on wound size

What is granulation tissue scaffolding ultimately converted to?

Scar

What kind of granulation tissue turns forms a pale, avascular scar?

Richly vascularised granulation tissue

What growth factors are involved in extracellular matrix (ECM) deposition during tissue repair?

Transforming growth factor β(TGF-β)
Fibroblast growth factor (FGF)
Platelet derived growth factor (PDGF)

What is the purpose of extracellular matrix deposition?

Formation of scars from granulation tissues

What is the main purpose of tissue remodelling after scar formation?

The scar is remodelled to increase its strength and contract it

How does wound strength increase during tissue remodelling?

By increased cross-linking of collagen, enlargement of collagen fibres, and replacement of collagen type III with type I.

What does the balance between ECM synthesis and degradation influence?

The remodelling of connective tissue and the size and nature of the scar.

What enzymes are responsible for ECM degradation during tissue remodelling?

Matrix metalloproteinases (MMPs), which includes collagenases, stromelysins, and gelatinases

Summarise wound healing

1. Induction of acute inflammation
2. Regeneration of parenchymal cells
3. Migration and proliferation of connective tissue
4. Synthesis of ECM protein
5. Remodelling of connective tissue
6. Collagenisation and acquisition of wound strength

What is healing by first intention (also called primary union)?

When a wound only involves the epithelial layer, only requiring epithelial regeneration, which is the principal mechanism of repair

What is healing by second intention (also called second union)?

When cell or tissue loss involves more than the epithelial layer (such as large wounds, abscesses, ulceration, and infarction), requiring a combination of regeneration and scarring.

Compare first and second intention wound healing

1st intention
- little to no tissue has been lost
- opposed edges
2nd intention
- tissue has been lost
- unopposed edges
- more intense inflammatory reaction
- more granulation tissue
- contraction of wound

Why is there a more intense inflammatory reaction in second intention than first intention?

Because there are more debris and exudate

Why are there larger amounts of granulation tissue in second intention than first intention?

In deep wounds where drainage to the surface is poor, granulation tissue grows from the base upward to fill the defect before surface closure can occur.

What is the determining factor of whether a wound heals by first or second intention?

The nature of the wound is the determining factor, rather than the healing process

What is the strength of the wound at week 1-2?

10% strength of unwounded skin

What happens to the strength of the wound for the 4 weeks after week 1-2?

Strength of the wound increases rapidly

After 3 months, what happens to the strength of the wound?

The rate at which wound strength increases slows down once the strength has reached 70-80% of unwounded skin

Which skin cells regenerate efficiently after superficial injury?

Squamous epithelial cells

Why might some structures of the skin, like hair follicles and sweat glands, not return after skin injury?

They are specialised structures that are destroyed in deep wounds

What distinguishes a GIT mucosal erosion from an ulcer in healing?

  • Erosion = loss of only partial thickness, mucosa regenerates in hours if cause is removed
  • Ulcer = loss of full thickness (often has damaged muscle and blood vessels), destroyed muscle can't regenerate, heals by scar, can cause distortion/obstruction
  • Which tissue forms a callus during healing?

    Bone

    Summarise healing in bone from a fracture

    1. Haematoma formation = a blood clot at the fractured site that acts as a framework
    2. Callus formation
    3. Callus and woven bone replaced by lamellar bone
    4. Bone remodelling

    What are the factors that can delay bone healing?

    Movement
    Interposed soft tissue (unwanted tissue between two structures)
    Gross misalignment (anatomically inappropriate bone healing)
    Infection

    Which organ has exceptional regenerative capacity via hepatocytes?

    Liver

    What determines whether the liver heals normally or forms cirrhosis?

    Whether architecture framework of the liver is preserved or not

    When does liver form fibrous scars or regenerative nodules from healing?

    When confluent cells are lost or when the architecture of the liver has been damaged

    What are the local host factors that modify the healing process?

    Infection
    Mechanical factors (early motion, pressure)
    Foreign bodies (sutures, fragments, bone)
    Size, location, and type of wound

    What are the systemic host factors that modify the healing process?

    Nutrition (vitamin C)
    Metabolic status (diabetes)
    Circulatory status (what the blood vessels bring to the injured site)
    Hormones

    What is a chronic wound and why does it happen?

    A wound that fails to heal normally and remains open for a long time.
    1. Mainly due to local ischaemia
    2. Systemic factors (internal, body-wide condition)

    Why does a deficient scar form?

    Deficient scar = not enough scar tissue/collagen
    Forms due to mechanical stress like coughing and vomiting

    Compare a hypertrophic scar and a keloid

    Both are formed due to excess collagen laid down during the healing process.
    Hypertrophic scars are local and usually regress overtime.
    Keloids form beyond wound boundaries and don't regress.

    What causes replacement of normal tissue by fibrosis?

    Persistent tissue injury

    Explain the process of replacing of normal tissue by fibrosis due to persistent tissue injury.

    1. Persistent tissue injury leads to chronic inflammation and loss of tissue architecture
    2.  Macrophages and leukocytes produce cytokines to heal
    3. Fibroblasts and myofibroblasts migrate and proliferate + collagen and other ECM proteins are deposited

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