Platelet Rich Plasma (PRP)

Autologous Platelet-rich Plasma

• What is plasma?– Fluid component of a person’s 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 person’s 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 patient’s 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, plasma-rich 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

1. Formation of tri-dimensional mesh (fibrin strand)or matrix….

*Platelets & Megacaryocytes vol.2 Dr J.Gibbins, M. Mahaut-Smith

2. Release of growth factors by the thrombocytes and

leukocytes….

3. Chemo-attraction or migration of

macrophages and stem cells…

4. Stem cells proliferation &

mitosis…

5. Stem cells differentiation…

(In addition ECM like fibronection,

vitronecton, thrombospondin…)

The 5 Major Steps In The Platelet Activation Process

Timelines in Wound Healing

Benefits of a-PRP reported in the “Healing Cascade”

Physiologic response: time

% W

ound

clo

sure

Haemostasis

Inflammation

Tissue regeneration

Physiologic response: time

% w

ound

clo

sure

FibrinPlts agrgg

vWF

LeukocytesPlts G. Factors

Chemo tactics & mitotic G. Factors

Extra Cell. Matrix synth. & Cell differentiation

G. Factors

By concentratin

g specific cells, wound healing time

can be shortened

significantly

Tissue regeneration

InflammationHaemostasis

Wound healing withPRP

Wound healing without PRP

Tissue remodeling

Tissue remodeling

Growth Factors Acting on “Healing Cascade”

Factor Name Principal Source EffectsPDGF aaPDGF bbPDGF ab

Platelet derived growth factors

Activated thrombocytes

Mitogenes of mesenchymal stem cells promote the synthesis of the extracellular matrix

TGF- alphaTGF- beta

TransformingGrowth Factors

Activated thrombocytes

Stimulation of DNA synthesis, proliferation of various types of cells. Favours the synthesis of collagen

IGF- IIGF- II

Insulin-likeGrowth 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 EndothelialGrowth Factor

Leucosytes & Endothelial cells

Stimulate angiogenesis & chemo-attraction 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, CD-W60, CD61, CD62P, CD63

physiological process (days)

% o

f wou

nd c

losi

ng

0

100

Journal of Oral & Maxillofacial Surgery, 2000; 58:45 Marx, Monteleone, Ghurani, Dr. Robert Marx, University of Miami

0 30

Healing with PRP

Control sample

physiological process (days)

Pat

ient

dis

com

fort

(pai

n)

0

10

0 30

Pain with PRP

Control sample

PAIN REDUCTIONWOUND HEALING

Visible effect in time of Healing and Discomfort (randomized study USA)

Advantages of A-PRP

• Tissue regeneration & rejuvenation: neo-collagenesis (TGFα & β), neo-vascularisation (EGF & VEGF), & extracellular matrix formation (PDGFαα & ββ & αβ) – 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 & WBC’s & 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 doctor’s rooms (no external

laboratory required)• Cost effectiveness: 1 Plasma kit (2 tubes) delivers 12+ ml A-PRP

RESEARCH & DEVELOPMENT

   

Cell separation

   

Autologous cell culture

   

Autologous stem cells culture

   

Cell differential

   

Tissue regeneration

   

Healing remodelling

DENTAL MEDICINE

   

Dental extractionDental implantation

DERMATOLOGY INTERNAL MEDICINE

GERONTOLOGY   

Cutaneous reconstruction and

transplantation

   

Ulcer and chronic wound therapy(e.g. after radio

therapy)   

Re-implantation of Autologous cells,

extemporaneous or cultivated in-vitro

SURGERY   

Cardio-vascular surgery

   

Abdominal surgery

   

Maxillo-facial surgery

   

Orthopaedic surgery

   

Plastic & cosmetic surgery/dermatology

   

Treatment of severe burns

Fields of Application of A-PRP

A-PRP Indications for dermatocosmetic1. 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

FILLERS

Wrinkles & volumes

correction

BIOSTIMULATION

MyCells

Architectural skin reconstruction/ reorganization

DERMOCOSMETICIS

PEELS

Renewal of the corneal layer

Germinative layer Stimulation

Dermis Stimulation

Hyperpigmentation removal

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 Growth factors & Protein Released by PLTPLT

Growth FactorGrowth Factor PRPPRP(Platelet Rich Plasma)(Platelet Rich Plasma)

PPPPPP(Platelet Poor Plasma)(Platelet Poor Plasma)

VEGF 220.4±78.1pg/ml 72.9±32.5pg/ml

EGF 269.1±117.5pg/ml 73.5±36.3pg/ml

PDGF-BB 2048.4±645.8pg/ml 308±125.8pg/ml

PRP preparation(1)PRP preparation(1) Collect blood from central vein of elbow10cc for each tube

PRP and PPP

Gel separator

Red blood cell

Centrifuge:Centrifuge:4000 G4000 G, 7min7min.. After After

centrifugationcentrifugation

Mycells Mycells tubetube

PRP preparation(2)PRP preparation(2)

PPP aspiration

PRP

PRP just before PRP just before injectioninjection

Aspiration of PRP

Aspiration and Blow of PRP

PRP after PPP aspiration

Sleeve insertion

Upper eyelidSubdermal injections 0.2ml each x 3.Total 0.6ml.

Lower eyelidSubdermal 0.2ml injections 1 cm apart. Massage evenly. Total 1-2ml.

CheeksSubdermal & intradermal injectionsLinear threading technique 0.2ml per injection. Total 3-5ml per side.

Naso-labial foldsSubdermal & intradermal injectionsLinear threading technique 0.2ml per injection. Total 2-3 ml per side.

LipsVermillion border injectionsLinear threading technique 0.2ml per injection. Total 0.4ml per quadrant.ChinLinear threading technique 0.2ml per injection. Total 2-3ml per side.

ForeheadIntradermal injections 0.05ml. Total for forehead 3ml.

KUBOTA JUNICHIRO CLINICKUBOTA JUNICHIRO CLINIC

• 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.

How do we inject PRP into the skin?How do we inject PRP into the skin?

PRP PRP injectioninjection

PRP just before injectionPRP just before injection

We usually inject PRP We usually inject PRP usingusing ““ linear injectionlinear injection ””and and ““mesotherapymesotherapy” technique over ” technique over the entire area to be treated.the entire area to be treated.

Inject as small spot(Inject as small spot(0.05cc to 0.05cc to 0.1cc0.1cc).).

Injecting layers are Injecting layers are intra-dermisintra-dermis and and subcutaneous tissuesubcutaneous tissue respectively.respectively.

30G needle 1cc syringe

Dec. 2006 March 2007

June 2008

6 time PRP injection

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