autologous cellular rejuvenation (acr)

5/19/2018 Autologous Cellular Rejuvenation (ACR)

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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 redblood cells


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What is Autologous Platelet-rich Plasma (a-PRP)?

A-Platelet rich plasma is a concentration of humanplatelets in a small volume of plasma measured as1,000,000 platelets per mm3 or 2-6 times the nativeconcentration 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 plateletconcentrate

PRP is also a concentration of the 7 fundamental

protein growth factorsthat have been proved to beactively secreted by platelets to initiate all woundhealing

PRP includes 3 proteins in blood known to act as celladhesion molecules: fibrin, fibronectin & vitronectin


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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


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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

collagenIGF- 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 epidermiscells,

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


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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


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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


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Timelines in Wound Healing


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Benefits of a-PRP reported in the Healing Cascade

Physiologic response: time

%Woundclosure

Haemostasis

Inflammation

Tissue regeneration

Physiologic response: time

%woundclosure

Fibrin

Plts agrgg

vWF

Leukocytes

Plts G. Factors

Chemo tactics

& mitotic

G. Factors

Extra Cell. Matrix synth.

& Cell differentiationG. Factors

By

concentrating

specific cells,wound

healing time

can be

shortened

significantly

Tissue

regeneration

Inflammation

Haemostasis

Wound

healing with

PRP

Wound

healing

without PRP

Tissue remodeling

Tissue remodeling


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physiological process (days)

%o

fwoundclosing

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)

Pa

tientdiscomfort(pain)

0

10

0 30

Pain with PRP

Control sample

PAIN REDUCTIONWOUND HEALING

Visible effect in time of Healing and Discomfort

(randomized study USA)


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Advantages of A-PRP

Tissue regeneration & rejuvenation: neo-collagenesis (TGF& ), neo-vascularisation (EGF & VEGF), & extracellular matrix formation (PDGF&& )NB: growth factors in genetically pre-determined ratio!

Bio-glue (fibrin glue): haemostasis & tissue adhesion in skin flaps, bonegrafts, 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 externallaboratory required)

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


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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)


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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


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A-PRP Indications

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 (Juvederm, Restylane,

Teosyal, etc) and calciumhydroxyl-appatite (Radiesse) = bio-active

filler containing growth factors

4. Acne scarring:

subscision injections & topical plasma & Dermaroller

5. Cellulite?

6. Striae (stretch marks)?


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SKIN TREATMENT ACTION LEVELS

MULTIDISCIPLINARY PROGRAM FOR THE REJUVENATION

OF THE FACE

FILLERS

Wrinkles &volumes

correction

BIOSTIMULATION

MyCells

Architectural skinreconstruction/

reorganization

DERMOCOSMETICIS

PEELS

Renewal of thecorneal layer

Germinative layerStimulation

Dermis Stimulation

Hyperpigmentation

removal


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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 Adaily for 7 days pre- & post-treatment

Radio-Frequency Immediately after treatment = platelet activation

Collagen fibre contraction (immediate)

Fibroblast induced neo-collagenesis (delayed)

Dermaroller & Topical A-PRP

micro-surgical needling

Induces growth factor release

Topical Vitamin A

Topical Vitamin C


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MyCells Important Factor

Presence of Progenitor CD34Bone Marrow

Mesenchymal Stem Cells in MyCells

Neocell Laboratories in Cape Town 2008

10 ml Venous blood sample yield 1.6 x 10CD34

stem cells

6 ml post centrifugation plasma yield 1.2 x 10CD34cells = 80% yield!


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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 perinjection. Total 3-5ml per side.

Naso-labial folds

Subdermal & intradermal injections

Linear threading technique 0.2ml per

injection. Total 2-3 ml per side.

LipsVermillion border injections


injection. Total 0.4ml per quadrant.

Chin


injection. Total 2-3ml per side.

Forehead

Intradermal injections 0.05ml. Total

for forehead 3ml.


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Side-effects

Minor oedema

Seldom bruising Eyelids can remain puffy for 2-3days

No infection

No allergy


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PRP preparation(1) Collect blood fromcentral vein of elbow

10cc for each tube

PRP and PPP

Gel separator

Red blood cellCentrifuge:2058G, 7min.

After centrifugation

Mycells tube

PRP ti (2)


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PRP preparation(2)

PPP aspiration

PRP

PRP just before injection

Aspirationof PRP

Aspiration andBlow of PRP

PRP after PPP aspiration

Sleeve insertion


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KUBOTA JUNICHIRO CLINIC

Now we are injecting the PRP using mesotherapylike 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?


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PRP injection

PRP just before injection

We usually inject PRP using

linearinjection and mesotherapytechnique over the entire area to betreated.

Inject as small spot(0.05cc to 0.1cc). Injecting layers are intra-dermisand

subcutaneous tissuerespectively.

30G needle 1cc syringe

0 f


24/25KUBOTA JUNICHIRO CLINIC

Case70 year old female

She was injected PRP around lower eyelid

and nasolabial fold.

Pre-injection of PRP 1 month after injection

Skin elasticity ,wrinkles and tension improved !!

She looks much younger than before.


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Dec. 2006 March 2007 June 2008

6 time PRP injection

autologous cellular rejuvenation (acr)

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