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Eq ACVS Phase 1 > Ch.8 Regenerative Medicine > Flashcards

Ch.8 Regenerative Medicine Flashcards

1
Q

What is PRP?

A

A plasma suspension containing a quantity of platelets that is increased in concentration from that of whole blood and also contains varying amounts of leukocytes, RBC and plasma proteins.

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2
Q

The use of PRP is based on what concept?

A

The delivery of growth factors and proteins of the clotting system may optimize tissue healing .

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3
Q

What is responsible for the antimicrobial properties of PRP?

A

Platelet lysates

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4
Q

What growth factors present in PRP are important in assisting bone and cartilage healing

A

Osteocalcin (Oc)
Osteonectin (On)
Fibronectin (Fn)
Thrombospondin-1 (TSP-1)

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5
Q

How soon after injecting PRP does the clotting cascade start?

A

Within minutes - 95% of the growth factors are secreted within 1 hour

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6
Q

Why is mutagenic activity and tumour formation not an issue with PRP application

A

It does not act directly on the nucleus

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7
Q

What is the half life of platelets

A

5-7 days

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8
Q

What should be considered when using combination therapy of suspending mesenchymal cells in PRP?

A

When platelet concentrates are supplemented in media at volumes >30% proliferation of mesenchymal stem cells declines.
Longterm suspension of mesenchymal cells in PRP may negatively affect mesenchymal cell viability

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9
Q

PRP preparation requires a “soft spin” and a “hard spin” why?

A

Soft spin - Leukocytes and RBC are larger and more dense than platelets so they settle faster leaving the platelets suspended in the plasma.

Hard spin - Centrifugation for platelets to be pelleted

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10
Q

What is P-PRP

A

Pure PRP or Leukocyte-poor
Contains platelets and plasma and little to no RBC or leukocyte. Produced by a soft spin where the RBC and leukocytes are pelleted and the plasma and platelets left suspended
eg Arthrex ACP Double syringe system

or in a lab by plasmapheresis

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11
Q

Why must Equine specific PRP kits be used to prepare equine PRP

A

Interspecies differences in blood viscosity, platelet density, degree of platelet activation and aggregation.

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12
Q

Effect of neutrophils, IL-1B and tumour necrosis factor alpha within tendons

A

Decreases matrix synthesis
Increases tissue catabolism

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13
Q

Effect of peripheral blood mononuclear cells PBMCs on repair

A

Enhance anabolic effects of PRP

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14
Q

How has IGF-1 been shown to affect tendonitis

A

Improved
1- cell proliferation,
2 - collagen content,
3 - mechanical stiffness and
4 - sonographic appearance during 8 weeks of tx

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15
Q

What is COMP (Cartilage oligomeric matrix protein) ?

A

A component of ligament healing response and a marker of appropriate ligament matrix remodelling.

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16
Q

What is the proposed use of acellular bone marrow aspirate?

A

Use instead of PRP on desmitis as
enhances healing by
providing anabolic factors
which stimulate matrix production

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17
Q

what component of PRP stimulates angiogenesis

A

VEGF
Vascular endothelial growth factor

18
Q

Up to how long post PRP tx is neovascularisation thought to occur

A

23 weeks

19
Q

2 primary pro inflammatory cytokines that initiate and participate in the destructive cytokine cascade of osteoarthritis

A

IL-1
TNF-Alpha

20
Q

How can the activity of IL-1 be inhibited

A

IL-1 receptor antagonist protein IL-1ra prevents the interaction btw IL-1 and its receptor

Direct injection of the antagonist
Gene therapy or conditioning of the serum to stimulate production of IL-1ra

21
Q

How is autologous conditioned serum ACS, produced?

A

Conditioning certain blood components, primarily monocytes, to increase endogenous production of IL-1ra

Accomplished by exposing whole blood to medical grade chromium sulphate treated glass beads.

22
Q

What increase in IL-1ra must occur to block all of the available IL-1 receptors?

A

10-1000 fold increase

23
Q

What is IRAP

A

(ACS)
InterLeukin1 Receptor Antagonist Protein
Orthokine - human product
Whole blood - medical grade chromium- sulphate treated glass beads - incubated at 37* for 24hrs, centrifuged, serum obtained and stored at -20*

6 2cc syringes given over 3 weeks

24
Q

Which system Arthrex or Dechra has been found to produce a high amount of IRAP

A

Arthrex

25
Q

What is APS, how is it produced?

A

Autologous Protein Serum
ProStride
20mins instead of 24 hours
Whole blood - APS separator sequesters WBC and platelets
Then transferred to an APS concentrator - filters it through Polyacramide beads and desiccates it
Result is a concentrated sol of WBC, platelets and plasma proteins

26
Q

How does APS differ to ACS?

A

Autologous Protein Serum

Higher levels of IL-1ra and platelet derived growth factors but also highly variable levels of IL-1B

27
Q

Where are stem cells most commonly harvested in equine medicine

A

Adipose - Tail head/sup gluteal area
Bone marrow - Sternum, ilium
Older horses >13 hard to harvest from ilium and less MSC than from sternum

28
Q

What methods can be used to prepare the stem cells

A

1 Culture and expansion
2 Patient side centrifugation BMC - Bone marrow wconcentrate

3 - If adipose derided need to be digested first - 3 days shipping and prep

29
Q

Non adult sources of stem cells

A

Fetal fibroblasts
Umbilical cord blood
Umbilical cord tissue/matrix
Placental tissue
Amniotic fluid and membrane

30
Q

What stage are embryonic stem cells derived from

A

blastocyst

31
Q

What are iPSCs induced pluripotent stem cells

A

Pluripotent stem cells generated through genetic reprogramming of adult somatic cells

Yamanka factors - mouse

32
Q

What are MSCs

A

Mesenchymal stem cells
undiferentiated cells capable of asymmetric cell division - one daughter cell develops into a somatic cell type while the other remains as a stem cell in the “niche” tissue

33
Q

How do MSCs aid in inflammation

A

Signal endothelial cells to promote angiogenesis
Prevent leukocyte transmigration
Stimulate fibroblasts to produce extracellular matrix
Activate and recruit tissue intrinsic stem/progenitor cells

33
Q

How do MSCs aid in inflammation

A

Signal endothelial cells to promote angiogenesis
Prevent leukocyte transmigration
Stimulate fibroblasts to produce extracellular matrix
Activate and recruit tissue intrinsic stem/progenitor cells
Inhibit T cell activation, B-cell function and dendritic cell maturation

34
Q

What is the ability of MSCs to modulate T-cell prolif and cytokine secretion dependent on

A

PGE-2

35
Q

Cautions against using allogenic MSCs

A

Loss of efficacy of MSC as destroyed by host before eliciting effect
Risk of adverse rxns

36
Q

How is the best quality and content of MSC obtained on aspirate

A

First 5cc aspirate

If need 60 such as for Bone marrow concentrate aspirate 5 to 10 from different sites/depths

37
Q

What is the location for sternal aspirate

A

4th and 5th marrow spaces

38
Q

Where is adipose tissue harvested for stem cells and how much

A

Subvcut Tail head/superior gluteal
15-20g

39
Q

BMP-12 Bone Morphogenic Protein 12 been shown to aid in the formation of what tissue

A

Tendon and ligament like tissue

40
Q

Which possesses a higher capacity for chondrogenic differentiation UCB-MSCs or BM-MSCs

A

UCB-MSCs
umbilical chord blood derived MSCs

Eq ACVS Phase 1 (72 decks)

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