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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