Platelet Rich Plasma (PRP)
Autologous Platelet-rich Plasma
What is plasma?
Fluid component of a persons blood
Contains platelets, white blood cells, stem cells, electrolytes,
enzymes, hormones, nutrients, anti-bodies, glucose, proteins, lipids
& albumin (powerful anti-oxidant), etc.
Why autologous?
Autologous means persons own (self donated) and not donated
from another person or from animal origin
Growth factors must be in genetically pre-determined ratios!
No risk of rejection and lower allergenic potential
How can A-PRP be obtained easily?
Venous blood sample is obtained from patients fore-arm
Centrifugation separates plasma & platelets & stem cells from red
blood cells
What is Autologous Platelet-rich Plasma (a-PRP)?
A-Platelet rich plasma is a concentration of human
platelets in a small volume of plasma measured as
1,000,000 platelets per mm3 or 2-6 times the native
concentration of whole blood at a pH of 6.5 - 6.7
(whole blood pH = 7.0 - 7.2)
Also referred to as autologous platelet gel, plasmarich growth factors (PRGFs) or autologous platelet
concentrate
PRP is also a concentration of the 7 fundamental
protein growth factors that have been proved to be
actively secreted by platelets to initiate all wound
healing
PRP includes 3 proteins in blood known to act as cell
adhesion molecules: fibrin, fibronectin & vitronectin
How are Platelets Activated?
Dermal collagen & exposed endothelial collagen
Arachidonic Acid (inflammation pathway)
Thromboxane A2 (inflammation pathway)
ADP
Thrombin (bovine has high allergenic potential)
Substrate bound ligands of Glycoprotein II a / III b
Vasopressin
Adrenaline (20% patients no receptor!)
CaCl2
Thermal: controlled heat (Radio-frequency)
Vibration via Vortex device
Cryo-activation
The 5 Major Steps In The Platelet Activation Process
4. Stem cells
proliferation &
mitosis
1. Formation of tri-dimensional
mesh (fibrin strand)or matrix.
3. Chemo-attraction or
migration of
macrophages and stem
cells
2. Release of growth factors
by the thrombocytes and
leukocytes.
(In addition ECM like fibronection,
vitronecton, thrombospondin)
*Platelets & Megacaryocytes vol.2 Dr J.Gibbins, M. Mahaut-Smith
5. Stem cells
differentiation
Timelines in Wound Healing
Benefits of a-PRP reported in the Healing Cascade
Wound healing
without PRP
% Wound closure
Tissue remodeling
Tissue regeneration
Inflammation
Haemostasis
Physiologic response: time
Wound healing
with
PRP
% wound closure
Tissue remodeling
Inflammation
Haemostasis
Fibrin
Fibrin
Plts
Plts agrgg
agrgg
vWF
vWF
Leukocytes
Leukocytes
Plts
Plts G.
Factors
Factors
Extra Cell. Matrix synth.
Tissue Extra Cell. Matrix
& Cell differentiation
regeneration & Cell
G.
G. Factors
Factors
Chemo
Chemo tactics
tactics
&
& mitotic
mitotic
G.
G. Factors
Factors
By
By
concentratin
concentratin
gg specific
specific
cells,
cells, wound
wound
healing
healing time
time
can
can be
be
shortened
shortened
significantly
significantly
Physiologic response: time
Growth Factors Acting on Healing Cascade
Factor
Name
Principal Source Effects
PDGF aa
PDGF bb
PDGF ab
Platelet derived
growth factors
Activated thrombocytes
Mitogenes of mesenchymal stem
cells promote the synthesis of the
extracellular matrix
TGF- alpha
TGF- beta
Transforming
Growth Factors
Activated thrombocytes
Stimulation of DNA synthesis,
proliferation of various types of
cells. Favours the synthesis of
collagen
IGF- I
IGF- II
Insulin-like
Growth Factors
Activated thrombocytes
Stimulates the proliferation and
differentiation of osteoblasts
EGF
Epidermal Growth
Factor
Activated thrombocytes
Stimulates proliferation and
differentiation of epidermis cells,
co-stimulating angiogenesis
VEGF
Vascular Endothelial
Growth Factor
Leucosytes &
Endothelial cells
Stimulate angiogenesis & chemoattraction of osteoblasts
In addition the activated thrombocytes have onto their surface a multitude of
signalisation molecules eg. CD9, CD-W17, CD31, CD41, CD42a-d, CD51, CDW60, CD61, CD62P, CD63
Visible effect in time of Healing and Discomfort
(randomized study USA)
WOUND HEALING
10
% of wound closing
Healing with
PRP
Control sample
Patient discomfort (pain)
100
PAIN REDUCTION
Control sample
Pain with PRP
0
0
physiological process (days) 30
physiological process (days) 30
Journal of Oral & Maxillofacial Surgery, 2000; 58:45 Marx, Monteleone, Ghurani, Dr.
Robert Marx, University of Miami
Advantages of A-PRP
Tissue regeneration & rejuvenation: neo-collagenesis (TGF & ), neovascularisation (EGF & VEGF), & extracellular matrix formation (PDGF &
& ) NB: growth factors in genetically pre-determined ratio!
Bio-glue (fibrin glue): haemostasis & tissue adhesion in skin flaps, bone
grafts, trauma intra-surgery and post-surgery
Safety: non-allergenic & free from concerns over transmissible diseases
e.g. HIV, Hepatitis B & C, CJD, etc.
Autologous: no risk of rejection reaction
Wound healing time: increased
Physiological anti-biotic : anti-bodies & WBCs & proteolytic enzymes
Plasma includes: hormones, bio-transformed vitamins & other nutrients
Tissue engineering: in-vitro autologous tissue culture-medium.
Ease of use: dermal & hypodermal injections
Convenience: harvesting performed in doctors rooms (no external
laboratory required)
Cost effectiveness: 1 Plasma kit (2 tubes) delivers 12+ ml A-PRP
Fields of Application of A-PRP
RESEARCH &
DEVELOPMENT
Cell separation
DENTAL MEDICINE
Dental extraction
Dental implantation
Cutaneous
reconstruction and
transplantation
Cell differential
Healing remodelling
Cardio-vascular
surgery
Abdominal surgery
Maxillo-facial surgery
Ulcer and chronic
wound therapy
(e.g. after radio
therapy)
Orthopaedic surgery
Plastic & cosmetic
surgery/dermatology
Tissue regeneration
Autologous stem cells
culture
SURGERY
Autologous cell culture
DERMATOLOGY
INTERNAL
MEDICINE
GERONTOLOGY
Re-implantation of
Autologous cells,
extemporaneous or
cultivated in-vitro
Treatment of severe
burns
A-PRP Indications for dermatocosmetic
1.
Skin rejuvenation:
injection (intradermis)
mesotherapy (intradermis)
topical plasma & Dermaroller micro-needling (intradermis)
2.
Fine lines & wrinkles: ditto
3.
Volumetric filling :
large volume injection of plasma intradermis and hypodermis of the tear
troughs, eyelids, naso-labial folds, marionette lines, peri-oral areas,
cheeks, forehead, glabella, neck & back of hands
A-PRP mixed with fillers such as hyaluronic acid (Esthelis, Juvederm,
Restylane, Teosyal, etc) and calciumhydroxyl-appatite (Radiesse) =
bio-active filler containing growth factors
4. Acne scarring:
subscision injections & topical plasma & skinroller/ skinneedling
5. Cellulite?
6. Striae (stretch marks)?
SKIN TREATMENT ACTION LEVELS
MULTIDISCIPLINARY PROGRAM FOR THE REJUVENATION
OF THE FACE
DERMO
COSMETICIS
PEELS
Renewal of the
corneal layer
Germinative layer
Stimulation
FILLERS
Wrinkles &
volumes
correction
BIO
STIMULATION
MyCells
Dermis Stimulation
Hyperpigmentation
removal
Architectural skin
reconstruction/
reorganization
Pre-Treatment Patient Preparation
& Combination Therapy
High Dose Oral Vitamin C (1,000mg+) daily for 7 days
pre-treatment & post-treatment
Enhances wound healing (fibroblast stimulation)
Oral Vitamin A daily for 7 days pre- & post-treatment
Radio-Frequency
Immediately after treatment = platelet activation
Collagen fibre contraction (immediate)
Fibroblast induced neo-collagenesis (delayed)
Skinroller & Topical A-PRP
micro-surgical needling
Induces growth factor release
Topical Vitamin A
Topical Vitamin C
Side-effects
Minor oedema
Seldom bruising
Eyelids can remain puffy for 2-3days
No infection
No allergy
Potential Complications
(applicable to all dermal fillers)
Intra-vascular injection (thrombus/embolus)
- Venous
- Arterial
Nerve trauma (needle)
Secondary infection
NB: Beware of the peri-ocular area (eyelids) - no
coagulant used eg. thrombin or CaCl2
Contra - Indications
Platelet Dysfunction Syndrome
Critical Thrombocytopenia
Hypofibrinogenaemia
Haemodynamic Instability
Auto-immune disease
Chronic oral steroid therapy
Chronic topical steroid therapy of treatment area
Malignancy
Chemo-therapy
Sepsis
Acute & Chronic Infections
Chronic Liver Pathology
Anti-coagulation therapy (warfarin, aspirin)
Pregnancy (for cosmetic indications)
Why MyCells?
Separator gel clear plasma
Glass tube non toxic
Complete sterilization process
Only 10cc blood needed
RCF larger plasma yield
Regulatory certificates
Simple & easy to use
Growth factors & Protein
Released by PLT
Growth Factor
PRP
(Platelet Rich Plasma)
(Platelet Rich Plasma)
PPP
(Platelet Poor Plasma)
VEGF
220.478.1pg/
ml
72.932.5pg/m
l
EGF
PDGF-BB
(Platelet Poor Plasma)
269.1117.5pg 73.536.3pg/m
/ml
l
2048.4645.8p
g/ml
308125.8pg/
ml
PRP preparation(1)
Collect blood from
central vein of
elbow
10cc for each tube
PRPandPPP
Gelseparator
Mycells
tube
Centrifuge:4000 G,
G
7min.
Redblood
cell
After
centrifugation
PRP preparation(2)
PRP
Aspiration
and Blow of
PRP
Sleeve
insertion
PPP
aspiration
PRP after PPP
aspiration
Aspiratio
n of PRP
PRP just before
injection
Forehead
Intradermal injections 0.05ml. Total
for forehead 3ml.
Upper eyelid
Subdermal injections 0.2ml each x 3.
Total 0.6ml.
Lower eyelid
Subdermal 0.2ml injections 1 cm
apart. Massage evenly. Total 1-2ml.
Cheeks
Subdermal & intradermal injections
Linear threading technique 0.2ml
per injection. Total 3-5ml per side.
Naso-labial folds
Subdermal & intradermal injections
Linear threading technique 0.2ml
per injection. Total 2-3 ml per side.
Lips
Vermillion border injections
Linear threading technique 0.2ml
per injection. Total 0.4ml per
quadrant.
Chin
Linear threading technique 0.2ml
per injection. Total 2-3ml per side.
How do we inject PRP into the skin?
Now we are injecting the PRP using mesotherapy like
technique over the entire area to be treated.
Inject small spot (0.05cc to 0.1cc) into the skin.
Injecting layer is dermis and subcutaneous
tissue.
KUBOTA JUNICHIRO CLINIC
PRP
injection
30G needle
1cc syringe
PRP just before injection
We usually inject PRP
using linear injection and
mesotherapy technique over
the entire area to be treated.
Inject as small spot(0.05cc to
0.1cc).
Injecting layers are intra-dermis
and subcutaneous tissue
respectively.
Dec. 2006
March
2007
June 2008
6 time PRP injection
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