Slide 1

PRP THERAPY : A MODERN

MEDICINE BOON FOR REPAIR,
RESTORATION AND
REJUVENATION
Dr. Suryakant Hayatnagarkar
Cryo Cell India Pvt. Ltd.
Cryo Cell

Platelet rich plasma therapy is gaining

importance recently. It involves use of platelet
derived factors to treat various diseases
involving almost all tissues and systems of
body.
A new method of treatment is always associated
with high expectations for its success. This is
more true when therapy is potentially based on
premise of achieving symptomatic relief as well
as improving functional quality and repairing
structurally damaged tissues. Platelet rich
plasma (PRP) application is shown to be
successful catalyst in the healing process for

wide varieties of conditions in animals and

human models. It is something equivalent to
Aksir ilaaj , Rambad upay, an allround wonder
therapy.
PRP was earliest used in human treatment in
1987 in an open heart surgery procedure. PRP
therapy began gaining popularity in the mid
1990s. Methods of harvesting and using platelet
rich plasma are now becoming widely practiced
in the fields of sports medicine, orthopedics,
traumatology, general surgery, dentistry,
dermatology and cosmetology.
Slide 2
PLATELETS

The ubiquitous tiny blood cells derived

from megakaryocytes in bone marrow

2-3 microns in size, devoid of

nucleus,DNA

Contain dense granules and alpha granules

Cryo Cell

Platelets are the smallest fragments of blood

cells derived from fragmentation of precursor
megakaryocytes in bone marrow. 2-3 microns in
size platelets are devoid of nucleus or DNA and
life span is also short 3-5 days. Platelets attract
media attention mostly during Dengue epidemic
when people realize their importance, as life
threatening platelet reduction is common in
severe infections and it then becomes critically
deficient resource.
Though media does not realize but we know
platelets are very important source for therapy
in wide variety of diseases requiring repair and
tissue healing.
Slide 3

PLATELETS

Cryo Cell

On a stained blood smear, platelets appear as

dark purple spots, about 20% the diameter of
red blood cells. The ratio of platelets to red
blood cells in a healthy adult is 1:10 to 1:20.
Slide 4
PLATELETS

Cryo Cell

The main function of platelets is hemostasis by

attaching to any breach in vessel where platelets
aggregate, activate and release granules. The
platelets organelle zone is rich in platelet
granules. Alpha granules contain clotting
mediators such as factor V, factor VIII,
fibrinogen, fibronectin, platelet-derived growth
factors, and chemotactic agents. Delta granules,
or dense bodies, contain ADP, calcium,
serotonin, which are platelet-activating
mediators
Slide 5
PLATELET DERIVED FACTORS

Tissue regeneration

Angiogenesis

Epithelialization

Proliferation and differentiation of osteoblasts

Synthesis of collagen

Cryo Cell

Activated platelets release granules by

exocytosis which contain not only clotting
factors but more than sixty different biologically
active substances that are involved in processes
of tissue regeneration, angiogenesis,
epithelialization, proliferation and
differentiation of osteoblasts, and in synthesis of
collagen
Slide 6
PLATELET DERIVED GROWTH
FACTORS

EGF

PDGF Platelet derived growth factor

IGF

Insulin like growth factor

VEGF

Vascular endothelial growth f

TGF

Transformative growth factor

Epidermal growth factor

Cryo Cell

Various factors present in platelet granules are

Slide 7
PLATELET DERIVED GROWTH
FACTORS

KGF

Keratinocyte growth factor

FGF

Fibroblast growth factor

CTGF

Connective tissue growth factor

TNF alpha Tumor necrosis factor

Various Interleukins

Cryo Cell

Slide 8
The Primary Growth Factors Role
in Soft Tissue Healing and Repair

PDGF
2 polypeptide chains
AB, BB and AA types (AB in human
platelets)
Stored in alpha granules in platelets
TGF beta
Also polypeptide, multiple chains, TGF beta-1
most studied form
Found in platelets and bone, cartilage
Cryo Cell

The characters of primary growth factors role in soft

tissue healing and repair are
PDGF
2 polypeptide chains
AB, BB and AA types (AB in human platelets)
Stored in alpha granules in platelets
TGF beta
Also polypeptide, multiple chains, TGF beta-1 most
studied form
Found in platelets and bone, cartilage

Slide 9

Effect of PDGF on Tissues

Immediate

(within 5 minutes)

Second messenger stimulation

Inflammatory response
Early

(30min to 4 hours)

M-RNA stimulation, protein synthesis

Chemotaxis (draws cells to the area)
Late

(4-24 hours)

Fibroblast mitosis
Cryo Cell

Effects of PGDF on tissues are

Immediate (within 5 minutes)
Second messenger stimulation
Inflammatory response
Early (30min to 4 hours)
M-RNA stimulation, protein synthesis
Chemotaxis (draws cells to the area)
Late (4-24 hours)
Fibroblast mitosis

Slide 10
Effect of Platelet Growth
Factors on Tissues

It appears that through a complex series of

biochemical and cellular events, these growth
factors cause a rapid and sustained increase in
the number of fibroblasts in an area and then
through a poorly understood interaction among
factors modulates cellular activity to cause stem
cell maturation.

Cryo Cell

It appears that through a complex series of

biochemical and cellular events, these growth
factors cause a rapid and sustained increase in
the number of fibroblasts in an area and then
through a poorly understood interaction among
factors modulates cellular activity to cause stem
cell maturation.
Slide 11

PRP THERAPY

What do these Growth Factors Do??

Chemotaxis
Directional movement in response to a chemical stimulus
Stems cells are attracted to the growth factors and

migrate into the area

Cell proliferation
Significant increase in cellular reproduction activity

Possibly even a systemc effect

Performance enhancing?

Cryo Cell

The PRP treatment using extract of platelet

derived growth factors is an attempt to mimic
the natural response of living organism to tissue
damage. Release of platelet granule derived
growth factors initiates a chain reaction of
chemotaxis, proliferation, cell differentiation
and synthesis of extracellular matrix. Thus PRP
therapy can be directed to not only to repair
damage but also to create a biological model
encouraging tissue regeneration. The most ideal
therapy for tissue regeneration is using stem
cells. PRP can do the same regeneration to
almost 90% extent at minimal costs and
minimal setup.

Natural ageing process of tissues weakens the

regeneration potential, which can be effectively
bypassed by using PRP, directed to enhance
regeneration.
Slide 12

The most
natural
injectable
Cryo Cell

Preparation of platelet rich plasma.

Preparation of PRP is basically two step
centrifugation process. First step is done at 1500
RPM after adding appropriate anticoagulant for
15 min. Plasma containing most of the platelets
is separated to another tube which is centrifuged
at 2500 RPM to prepare platelet pellet.

Appropriate quantity of plasma is left behind to

resuspend the pellet.
This makes non activated PRP. It is not
necessary to specifically activate as injection in
tissue is sufficient to stimulate platelets to
release growth factors slowly.
Slide 13
Definition:
BUFFY COAT =PLATELETS + LEUCOCYTES

Platelet-rich plasma (PRP) is blood plasma that has

been enriched with platelets.
As a concentrated source of autologous platelets, (17-19
folds / 3.3million /mm3)
PRP contains ,and releases (through degranulation)
several different growth factors and other cytokines that
stimulate healing of bone and soft tissue.
Cryo Cell

Slide 14
TYPES OF PRP
P-PRP

Pure platelet rich plasma

L-PRP Leucocyte and Platelet rich plasma
P-PRF
Pure platelet rich fibrin
L-PRF
Leucocyte and platelet rich fibrin

A-PRP

Activated platelet rich plasma

Cryo Cell

Nomenclature of PRP is varying depending on

methods used in preparation. There is lot of
confusion about nomenclature and use of
specific product in literature.
The current consensus classification of PRP
preparations is based on a simple classification
system dividing so many types of products in 4
main families, based on their fibrin architecture
and cell content: Pure Platelet-Rich Plasma (PPRP), Leukocyte- and Platelet-Rich Plasma
(LPRP), Pure Platelet-Rich Fibrin (P-PRF),
Leukocyte- and Platelet-Rich Fibrin (L-PRF).
The 4 main families of products present

different biological signatures and mechanisms,

and obvious differences for clinical applications.
This classification serves as a basis for further
investigations of the effects of these products.
If required PRP can be activated using calcium
ions which makes it to a-PRP. There is a chance
of strong clotting which may make it unsuitable
for injection through insulin syringe.
a-PRP can be further incubated at 37C to
prepare expressed serum containing all the
growth factors, which can be used devoid of the
clot.
Another important preparation is Platelet rich
fibrin, in which fibrin clot is prepared which is
rich in platelets. The clot then is compressed to
make a membrane which can be used in dental
reconstruction or bone reconstruction. Fibrin
provides good scaffolding matrix for growth of
regenerating cells.
Slide 15

USE OF PRP IN VARIOUS

DISCIPLINES
PRP therapy finds usage in most
of the medical disciplines and
scientists have used PRP in
treating varied diseases
requiring rapid repair and
restoration.
Cryo Cell

PRP being almost an all round therapy it can be

used in almost all the disciplines of medicine.
Every faculty of medicine can be benefitted use
of PRP for treatment of specific conditions
requiring repair, regeneration and regeneration.

Slide 16
DENTISTRY
Dental

Implants
Alveolar ridge restoration
Sinus grafting and guided
bone regeneration
Cryo Cell

You must be wondering why am I mentioning

Dentistry first while taking about PRP uses. But
it is must to give the credit to Dentistry for using
PRP therapy on larger scale for guided
regeneration of bone tissue. Utilizing the
methods of enhancement of bone and soft
tissues regeneration using PRP are being used in
periodontics, oral implantology and oral and
maxilla-facial surgery.
New bone formation is a prerequisite for
regeneration of tissues lost through periodontal
diseases and for osseo-integration of implants
used in restorative dentistry.

In implant surgery most frequent hindrances at

the implantation site are lack of adequate bone
availability, proximity of maxillary sinus and
inferior alveolar nerve canal. Advanced
procedures like sinus grafting and guided bone
regeneration are necessary is many cases before
implants can be securely placed. Use of PRP
enhances the results of procedures like sinus
lifts, ridge augmentations multifold at the same
time enhancing potential of osseo-integration of
implants. All cases where hydroxyapatite bone
graft powder material is used, combination with
PRP enhances the bone formation many folds
due to growth factors supplied by PRP.
In most of the maxillofacial surgery procedures
use of PRP results in increase in healing speed,
two to three folds thus reduces the healing time
and offers hastened restoration of function.
Slide 17

ORTHOPEDICS

Bone

regeneration
Soft tissue functional
restoration
Cryo Cell

Orthopedics had direct advantage of PRP use

based on the experience gained by dentistry.
Uses in Orthopedics are varied. Roughly can be
divided into bone regeneration uses and soft
tissue function restoration uses.
Slide 18

BONE REGENERATION
All

open and internal reduction fixation

operations
Spinal surgeries
Scoliosis Repair
Spinal Fusion
Joint replacement surgeries
Total Hip Replacement
Cryo Cell

Total Knee Replacement

There is conclusive proof that PRP helps in non

union fracture treatment. When surgical
procedures are performed along with use of PRP
the healing is early and sure. In practical studies
It was found that platelet derived growth factors
were deficient in all non-union cases and when
restored with PRP the healing happened in
average 8 weeks.
Autologous bone grafts are used for spinal
fusion surgeries. When used in combination
with PRP it resulted in goon union in all the
cases.
Use of PRP as adjunct in joint replacement
surgeries is very interesting study. In a recent
review in literature where PRP was used in 98

cases of various joint replacement surgeries,

PRP was applied intraoperatively to exposed
tissues, synovium and lining of wound at the
closure. The patients had less postoperative
blood loss, fewer pain reducing medications,
greater range of movements at the discharge and
shorter hospital stay compared to those who did
not have PRP applications. This study directly
concludes that direct application of PRP to
operative site after TKR seals the tissues and
enhances integration of the implant material.
With newer press fit type implants which avoid
use of bone cement use of PRP promotes earlier
and more complete osseo-integration of
implants into host bone.
Slide 19

BONE REGENERATION

Surgeries requiring bone grafting or artificial

bone graft use.
Hand and Foot surgeries

Cryo Cell

PRP should be used in combination with

autograft, allograft, demineralized bone matrix,
hyxroxyapatite powder or any other graft
material used to fill bone defects. If the graft
material is used in form of tiny chips or powder
it is best to use PRP to mix the graft material
along with activator so that graft material and
PRP can form a firm gel which can be filled in
the defect in form of putty which will provide
three dimentional matrix as well as all necessary
growth factor for enhanced healing and repair.
Slide 20

SPORTS INJURIES

Sports injuries are very common and require

intensive therapy to restore the function of
injured part hundred percent.
The repair and restoration must be very quick as
long treatment waits will result in longer
rehabilitation periods and monetory loss to the
sportsperson.
Cryo Cell

Injuries in various sports are very common and

become a media news. The treatment and
rehabilitation has to be quick, and prompt and
effective therapy is expected and all these things
get discussed in media regularly. In sports
medicine PRP therapy must be enhancing the
economic potential of sportspersons by billions
of rupees.
PRP became a big news when high profile
sports persons were treated with.
Slide 21

Real-Life Examples

Many athletes use PRP Therapy

Rafael Nadal
Tiger Woods
Hines Ward
Kobe Bryant
Brandon Roy

Cryo Cell

Most of these atheletes use PRP quite often for

their injuries.
Slide 22
Aesthetic Medicine
In USA PRP therapy used for Asethetic
Skin Treatment is termed as Dracula
Therapy
It can be used as Anti-aging Vampire
Fascial and used for removal of wrinkle
lines and making skin younger looking.
Cryo Cell

In USA PRP therapy used for Asethetic Skin

Treatment is termed as Dracula Therapy

It can be used as Anti-aging Vampire Fascial

and used for removal of wrinkle lines and
making skin younger looking.

Slide 23
SKIN REJUVENATION
Face

and neck
revitalization
Winkle lines reduction
Under eye bags
Cryo Cell

Skin ageing is an unpreventable, irreversible

process that is influenced by both intrinsic and
extrinsic factors. The intrinsic factors like the
reactive oxygen species along with matrix
metalloproteinases (MMP) reflect different
physiological and pathological processes
involved in skin ageing. There is accumulation
of fragmented collagen fibrils that prevents neocollagenesis and causes further degradation of
the extracellular matrix (ECM).
Conventional anti-aging strategies, including
lasers and topical treatments, increases ECM
synthesis through the activation of fibroblasts.

Usage of PRP in aesthetic medicine is a new

concept. Advantages of using PRP in aesthetic
medicine include, tissue regeneration and
rejuvenation, induction of cell differentiation,
extracellular matrix formation, recruitment of
other cells to the site of injury and an increase in
collagen production. This can increase skin
thickness and overall skin health.
Platelets release growth factors and cytokines
locally which facilitates cell stimulation,
proliferation, differentiation and extracellular
matrix accumulation. This reverses Ageing
process which is due to reduction and
degradation of ECM and reduction of collagen.
When PRP is injected in dermis of damaged
area, it stimulates the tissue, causing mild
inflammation that triggers the healing cascade.
As a result new collagen begins to develop. As
the collagen matures, it begins to shrink and
tightens and strengthens the skin, as well as the
tendons and ligaments of the damaged area
when injected at that level.

Improvements in skin texture and tone are

noticeable within 3 weeks. Full collagen
regeneration requires 3 months.
Advanced wrinkling cannot be reversed also
severe scarring does not respond to treatment
which needs to be resected.
Slide 24
Androgenic Alopecia
Hair

follicle rejuvenation
Early hair loss treatment
Hair Transplantation
procedures
Cryo Cell

Androgenetic alopecia (AGA) is the most

common form of hair loss but current treatment
options are limited and moderately effective.
Efforts are on to understand cellular pathways
and molecular mechanisms involved in the
pathophysiology of alopecia, so as to target

potential treatment that not only stimulates hair

growth, but induces formation of new hair
follicles.
Current strategies for treatment of pattern hair
loss mainly focus on promoting cellular
proliferation and differentiation during hair
growth cycle. Minoxidil is such agent which
promotes survival of dermal papilla cells.
Finesteride reduces apoptosis of hair follicle
cells. Both the medicines work on very limited
basis. PRP which provides a battery of growth
factors can do this job more efficiently. If use
properly in early stage of hair-fall, it can work
wonders and can reverse the process of hair loss
completely.
EGF & FGF activates the proliferation & causes
trans-differentiation of hair stem cells &
produce new follicular units. bFGF & betacatenin, promotes the anagen phase of papilla
cells & thereby plays a key role in elongating
hair shaft. The antiapoptotic regulators, Bcl-2
protein & Akt signaling, that are activated by
these GFs, prolongs the survival of dermal

papilla cells during the hair cycle. Thus, the

mitogenic & antiapoptotic effects, of PRP
prolong survival of dermal papillae. PRP
modulates angiogenesis and enhance blood flow
around hair follicles, thus improving cutaneous
ischemic conditions.
The use of PRP mesotherapy either alone or as
an adjunct to surgical procedures in the patients
of androgenic alopecia thus holds promising
results.
PRP can be injected as inter-follicular injection
as 0.1 ml/cm2, in a retrograde fashion from deep
to superficial, at every centimeter, throughout
the treated site while in
mesotherapy the microneedle roller of 1-mm
fine needles is rolled over scalp followed by
interfollicular injection of PRP over the treated
area.
It is also used as an adjunct to hair
transplantation to increase the survival rate after
implantation. The pretreatment of follicular
units with PRP before transplantation has
resulted in improved hair growth and density.

The hair follicle is dipped into PRP for 15

minutes, before implantation and after transplant
PRP is injected into both the donor and recipient
area of scalp to minimize bleeding, stimulate
wound healing & to reduce scarring .
Though, PRP is a promising newer technique in
a dermatosurgeon's armamentarium but it is yet
to show consistency in results.
Slide 25
Advantages of PRP therapy

Autology: your own blood

Tissue regeneration and rejuvenation
Safety: non-allergenic and free from concerns over transmissible
diseases such as HIV, Hepatitis B & C, CJD and so on
Faster healing: no fibrosis formation compared to some long term
fillers, for example
Easy to use: dermal and hypodermal injections require only one
syringe and one needle
Convenience: PRP harvesting can be easily and quickly performed in
clinic. A sterile and registered specific kit for the blood puncture, the
preparation of the plasma and the materials is used.
Cost effectiveness

Cryo Cell

Advantages of PRP rejuvenation are

Uses bodys own natural platelets so there is no
risk of allergic reaction

Natural collagen formation in response to

presence of activated platelets
PRP is ideal for patients who do not want any
synthetic fillers
There is very little swelling, bruising, lumping
as fluid assimilates in natural skin environment
PRP can be used to enhance laser procedures for
faster and improved healing.
PRP therapy is equally effective in men and
women.
Slide 26
MISCELLANEOUS USES

Sinus and fistula surgery

Atrophic vagina syndrome
Adjunct therapy for wound healing
For sternal wound healing in cardiac surgery

Cryo Cell

PRP can be practically used in any case of

delayed healing or where healing process gets

hindered due to any reason. There are suggested

many uses of PRP in literature. Few such
conditions are like sinus and fistula surgery
I have tried to cover as much as possible
literature review to bring this small presentation
and interesting possibilities of participation of
laboratory in actual treatment of patient. With
your cooperation many such possibilities can be
explored to enhance the cure for patients which
will directly give benefits to hospitals instituting
such therapies.
Thank you.
Slide 27
CHRONIC SKIN ULCERS
Diabetic

chronic wounds
Diabetic foot
Decubitus ulcers, Pressure sores
Chronic non-healing ulcers
Ulceration due to varicose veins
Cryo Cell

Any wound goes through stages of acute

inflammation, increased mitotic activity and
finally orderly repair. Chronic wounds can be
those wounds failing to proceed through an
orderly and timely process to produce anatomic
and functional integrity. Practically, a chronic
wound is the one which has failed to heal within
3 months.
Repeated trauma, foreign bodies, pressure
necrosis, infection, ischemia and tissue hypoxia
are few of the reasons of non healing. Growth
factors are crucial for timely wound healing.
Platelets can fill the gap by supplying growth
factors. It is desirable to supply growth factors
over extended time and this can be achieved by
delivering platelets in fibrin matrix.
Fibrin membranes are very useful for treating
chronic diabetic non healing ulcers, and
pressure sores. Use of fibrin membranes in such
cases can prepare the wound for skin grafting
earlier, reducing the time for total healing. This
results in improved quality of life and lower cost
of care over other conventional therapies.

Slide 28

Thank You!

Cryo Cell

I tried to give a theoretical overview of PRP and

its use in Dermatology and Aesthetic medicine.
Practical application and further research
depends on cooperation between you and me.
As with any new therapy there are always
doubts about its effectivity in minds of
clinicians as well as patients. Most of the new
therapies that are not based on sound scientific
principles come and go as fashion.
But PRP is such a unique therapy which is
based on sound scientific principles and also
shows promising success in initial trials. Now it

is upto us to develop the therapy into sound

treatment choice by using therapy in various
indicated conditions.