This document discusses regenerating facial tissues through platelet-derived growth factor (PDGF) injections rather than surgical facelifts. It provides calculations showing that the number of PDGF molecules released during injections is sufficient to stimulate the fibroblasts in the treated facial areas. Research studies cited found that PDGF injections led to angiogenesis within a week, maximum fibroblast activation within a month, and new collagen formation within two months, regenerating skin tissues. The technique aims to biologically rejuvenate facial tissues rather than just achieving aesthetic youth through damaging surgical procedures.

2. Vol.2 May 2010 N°1
Full Face Regeneration:
theoretical and practical protocol
Maurizio Ceccarelli (International Centre for Study and Research in Aesthetic and Physiological
Medicine – Roma)
J. Víctor García (Sociedad Española de Medicina y Cirugía Cosmética – Barcelona)
Premise
Ageing causes a reduction in facial volume and texture of the tissues that induces a fall of such for
gravity and the consequent appearance of a imperfection.
Cosmetic surgery, with the aid of lifting, removes a portion of the tissues to restore themselves to a
normal state. It is obvious that this is not a physiological intervention, but only a cosmetic treatment.
The tissues are damaged and the itself intervention, with fibrous internal induced, worsens the
microcirculation and aging more biologically the tissues. We have a young face aesthetically but old
biologically.
The physician, however, is concerned to regenerate tissues and inducing the initial appearance altered
by presenting a true biological rejuvenation. The scientific rationale of this operation belongs to the
Regenerative Medicine, a new medical branch derived from the Physiological Medicine.
The regeneration of tissues of the face is defined by the term Medical Face Lifting and today
proposes itself as an alternative to facelift surgery.
The Medical Face Lifting provides:
• The regeneration of skin tissue
• The regeneration of zygomatic and malar bone
• The regeneration of face adipose tissue
Regeneration of skin tissue
This intervention uses dermo-epidermal cells biostimulation with platelet growth factors. The
technique, developed in 2003 by Prof. J. Víctor García Universitat Autònoma de Barcelona in Spain, has
now spread throughout the world for major clinical responses subsequent to treatment.
For some time, the use of platelet growth factors is offered in various fields of medicine. Numerous
scientific publications report the results in dental, orthopedic surgery, maxillofacial surgery and
plastic surgery. Have recently been proposed works also in the rejuvenation of the skin.
Among the first to create protocols for separation and use of platelet growth factors have the dr.
Anitua who has published numerous works over time. All scientific papers published about the use of
platelet growth factors applied topically to the lesion. For a correct clinical response following topical
administration, it requires an increase in platelet concentration of 5 -10 times than normal. Indeed, using the Platelet

3. Rich Plasma (PRP) obtained after centrifugation. After centrifugation draw in the lower portion of the
plasma (1/3 less), where platelets are more concentrated.
Victor Garcia, Autonomous University of Barcelona in Spain has proposed, for the first time in 2003,
the use of platelet growth factors in the dermal (skin biostimulation). Obviously what was designed
for topical application would be reviewed in light of the introduction by injection into the dermis.
Deepen our understanding of the platelet growth factors. Growth factors have been discovered in
1986 by Professor Rita Levi - Montalcini and Prof. Stanley Cohen. These two scientists have received,
for this discovery, the Nobel Prize. Since 1986 much has been said about the growth factors. Today if
we do a search on Google 121,000,000 items appear.
The Platelet Derived Growth Factor
Growth factors are small protein fragments, belonging to the group of cytokines, capable of binding to
membrane receptors to activate or inhibit cellular functions. Can be produced by many cells and
tissues: platelets, fibroblasts, osteoblasts, skin cells, liver, kidney, lachrymal glands, etc..
The works by stimulating PDGF receptor tyrosine kinase. This induces the hydrolysis of
polifosfoinositoli. It frees the 1-3-difosfoinositolo that activates a receptor in the endoplasmic
reticulum. It has an entrance of calcium ions that activate protein kinase C. This leads to metabolic
activation of fibroblasts.
Deepen our knowledge of PDGF. This is, among other factors, in the alpha granules of platelets. PDGF
plays a specific action on the skin matrix synthesis and fibroblast activation.
Platelets contain two types of granules. The alpha granules capable to release PDGF, PF4, Fatt. V, Fatt.
XIII, fibrinogen and dense granules able to release serotonin, ADP and calcium. The release of the
factors contained in the granules following activation of platelets.
The lesion endothelial connects platelets with extravasal collagen. This leads to binding the platelet
with Von Willebrant factor and subsequent to platelet aggregation.
The platelets, in the activation process, join to connective collagen by Von Willebrant factor and
together with the fibrinogen. From this initial process follows the coagulation cascade.
On the effect of fibroblasts biological activity, platelet induced, a paper published in J. Biol Chem in
July 2008, tells us that:
- Even with only 5 ng/ml we have a strong induction of fibroblast proliferation.
And a paper published in Blood in August of 1984 tells us that:
- The concentration of PDGF in a healthy human serum after platelet degranulation, is approximately
20 ng/ml PDGF and half-life is very short, about 2 minutes.
We can then calculate:
- During platelet degranulation are released approximately 20 ng/ml of PDGF which corresponds to 20
x 10-9
g/ml;
- To get the corresponding number of moles, we divide the grams by the molecular weight of the
substance;
- Molecular weight of PDGF is 300,000 (3 x 105
);
- Divide the grams for PM (20 x 10-9
: 300,000) and get 6 x 10-14
moles;
- The number of molecules in one mole of substance is given by Avogadro's number (6 x 1023
);
- Multiply this number by the number of moles of released PDGF get the value of 36 x 109
molecules.
Of particular interest is the work of 2008 published on Communicative & Integrative Biology, where
we read:
- After 6-8 hours by a fibroblast stimulation with PDGF we have a recruiting tyrosine kinase receptors
on the cell surface and a second concentration of PDGF stimulation induces a greater response, more
important.
Then, after eight hours from a first bio-stimulation there is an increase of tyrosine kinase receptor.
Then, a second stimulation leads to an increased response of connective tissue cells.
We continue our calculations:
- In our biostimulation intervention treat, normally, face and neck.

4. - Develop the average volume of these areas by multiplying an area of 30 to 20 cm with a thickness of
2 mm (dermis). We get the value of 12 x 1012
micron3
.
- Now, considering that a cell has a volume of 100 micron3
, and the fibroblasts are 1% of dermal
volume, we can calculate the number of fibroblasts present in our treatment volume.
- Then, by dividing the total volume (12 x 1012
micron3
) for cell volume (100 micron3
) and 100 (1%) get
an number of fibroblasts equal to: 12 x 108
fibroblasts.
Now we have to calculate whether the available molecules of PDGF can activate the calculated
fibroblasts.
- We divide, therefore, the molecules of PDGF by the number of fibroblasts: 36 x 109
molecules: 12 x
108
fibroblasts.
- We get 30 molecules/fibroblast for milliliter of plasma injected. Since we inject, on average, 5 ml
per biostimulation session, we get: 50 molecules of PDGF/fibroblast.
But how many receptors are present on the cell wall of a fibroblast?
We know that cell surface molecules (Cluster of Differentiation) have been recorded to date in 350
different types (International Workshop and Conference on Human Leukocyte Differentiation
Antigens) and various CDs were collected in families of receptors.
The CDs that are stimulated with PDGF are tyrosine kinase receptor, a sub family.
In particular we classify:
1. Intracellular receptors and transmemebrane receptors.
2. The transmembrane receptors are divided into ionotropic and metabolotropic.
3. Metabolotropic receptors are divided, in turn, into:
· Receptors linked to G proteins
· Receptor coupled to kinase
· Receptor of guanylate cyclase
· Receptor of tyrosine kinase
Each family and subfamily has several types of receptor for a grand total of 350.
It is obvious that not all cells (in this case, fibroblasts) have all the families of membrane receptors.
Suppose that the fibroblast may have 10-20 different kinds of receptors and that one of these is a
tyrosine kinase receptor.
- We can divide the overall number of receptors (350) for 10 or 20. We get a number 35 or 18 for
each receptor type and thus a maximum of 35 tyrosine kinase receptor for fibroblast. 35 receptors to
stimulate with ... ... 150 molecules of PDGF!
The first clinical work and basic research of skin bio-stimulation with PDGF, made by Prof. J. Victor
Garcia and Dr. Antonio González-Nicolás gives us important histological information.
- After 7 days of biostimulation with PDGF we have a maximum of angiogenesis.
- After a month we have the maximum of activated fibroblasts.
- After two months we have a neocollagenogenesi of reticular collagen (type III) and a really skin
rejuvenation.
- Then, by dividing the total volume (12 x 1012 micron3) for cell volume (100 micron3) and 100 (1%) get
an equal number of fibroblasts: 12 x 108 fibroblasts.
Now we have to calculate whether the molecules of PDGF at our disposal can activate fibroblasts
calculated.
- We divide, therefore, the molecules of PDGF by the number of fibroblasts: 36 x 109 molecules: 12 x
108 fibroblasts.
- We get to 30 molecules per milliliter of plasma fibroblast injected. Since we inject, on average, 5 ml
per session biostimulation, we get: 50 molecules of PDGF per fibroblast.
But how many receptors are present on the cell wall of a fibroblast?
We know that cell surface molecules (Cluster of Differentiation) have been recorded to date in 350
different types (International Workshop and Conference on Human Leukocyte Differentiation
Antigens) and various CDs were collected in families of receptors.
The CDs are stimulated with PDGF receptor tyrosine kinase of a sub.

5. In particular we classify:
1. Receptors and intracellular receptors transmemebrana.
2. The transmembrane receptors are divided into ionotropic and metabolotropici.
3. Metabolotropici receptors are divided in turn into:
· Receptors linked to G proteins
· Coupled receptor kinase to
· Receptor guanylate cyclase
· Receptor tyrosine kinase
Each family and subfamily has several types of receptor for a grand total of 350.
It 'obvious that not all cells (in this case, fibroblasts) have all the families of membrane receptors.
Suppose that the fibroblast may have 10-20 different kinds of receptors and that one of these is a
receptor tyrosine kinase.
- We can divide the overall number of receptors (350) for 10 or 20. We get a number 35 or 18 for
each receptor type and thus a maximum of 35 receptor tyrosine kinase for fibroblast. Receptors to
stimulate with 35 ... ... 150 molecules of PDGF!
The first clinical work and basic research of bio-stimulation with PDGF skin, made by Prof. J. And Dr.
J. Victor Garcia Antonio González-Nicolás gives us important information of histological.
- After 7 days of biostimulation with PDGF have a maximum of angiogenesis.
- After a month we have the maximum of activated fibroblasts.
- After two months we have a neocollagenogenesi collagen lattice (type III) and organic skin
rejuvenation.
Summary
• The contact of platelets with connective tissue leads to the activation of these and to the
degranulation of alpha granules.
• PDGF released from alpha granules is 20 ng/ml and that of 36 x 109
molecules per milliliter.
• In a normal bio-stimulation with PDGF we have to put 12 x 108
fibroblasts. Each fibroblast has
an average of 30 tyrosine kinase receptor on his wall.
• For this fibroblast activation we have 30 molecules of PDGF per plasma milliliter.
• After 6-8 hours from the PDGF activation there is a recruitment of new receptors on the wall
of the fibroblast.
• A new stimulation with PDGF (30 ng/ml), 6-8 hours after, allows for a greater clinical response.
• After 30 days of PDGF have the maximum peak number of activated fibroblasts.
• Fibroblastic activation follows the neoformation of collagen type III (reticular)
From the above we propose a variation of the bio-stimulation with PDGF scheme:
• Collect the sample according to the scheme and the classic kit approved;
• Separate all the plasma and stir to mix the platelet;
• Conduct an initial biostimulation on the face of the patient;
• Wait 6-8 hours to allow the recruitment of fibroblasts receptor;
• Make a second biostimulation with Platelet Rich Plasma (lower third of the centrifuge tube)
After 30 days, we make.
• a biostimulation with a Medical Device containing aminoacids and a buffer. The precursor
aminoacids allow the neoformation of dermal matrix and the bicarbonate buffer maintains a pH
of 7.4 to avoid the formation of collagen I type.
• a epidermis regeneration by abrasion, peeling, PDGF powdering and occlusion
• an infiltration of wrinkles with APF (Autologus Plasmatic Fibrin) as a basis for fibroblasts
movement. The fibroblast proliferation under the wrinkle allows a dermal regeneration with
local reduction of the aesthetic problem.

6. The synthesis of fibrin represents the process of terminal blood coagulation cascade. To the vascular
injury following before the formation of platelet thrombus and after the formation of fibrin thrombus
by conversion of fibrinogen into fibrin.
Thrombin (derived from prothrombin) acts on transforming fibrinogen into fibrin which is then
stabilized by factor XIII. Thrombin cleaves fibrinogen B terminals allowing polymerization of
monomers that.
For the formation of thrombin is important the platelets presence. These bind on their surface the
VIII factor, V factor and the X factor enabling the transformation of prothrombin into thrombin.
The clot retraction follows from this and subsequent fibrinolysis for work of plasmin. The formation
of plasmin is almost contemporaneous with the formation of fibrin and follows the activation of
plasminogen by an activator secreted by the endothelial cells.
There are related mechanisms that regulate the activation and inhibition of synthesis of plasmin.
As mentioned, the main stimulus to the formation of plasmin following the release of a endothelial cells
factor. This indicates that the vessel was repaired and that the clot can be removed. The
transformation of plasminogen to fibrin occurs only for molecules incorporated into fibrin and not for
those in the circulation.
The formed plasmin degrades the fibrin clot forming fibrin degradation products (FDP). The molecules
of plasmin in the fluid phase are degraded to fibrinogen, V factor and VIII factor.
The action of plasmin widens even, indirectly, to the extracellular matrix hydrolyses. Indeed, plasmin
activates dermal metalloproteinase (collagenase, gelatinase, stromelinasi).
To allow a clinical response with autologous plasma fibrin we must increase the duration time of this
and therefore reduce the clot retraction and plasmin fibrinolysis.
The clot retraction is when shortening of the fibrin strands and squeezing the blood serum outside the
grid formed by these filaments. The retraction of the clot occurs through the work of activated
platelets that undergo a viscous metamorphosis, which melt and are the key elements of the formation
of fibrin. At the same start the production of a substance called reactozima, which stimulates the
retraction of the clot.
To prevent the clot retraction we separate, after centrifugation, the plasma from the cells.
Furthermore, to prevent fibrinolysis, add to plasma tranexamic acid.
Tranexamic acid prevent the binding of plasminogen to fibrin and this would therefore prevent
fibrinolysis. A drop of commercial product based on tranexamic acid (Tranex) contains 1018
molecules
of active ingredient, more than enough to act on the molecules of plasminogen in 10 ml of plasma (1017
molecules).
Autologous plasma fibrin is infiltrated in the dermis under wrinkles and/or depressions of the face.
Regeneration of bone tissue
The second step is the regeneration of bone. The soft tissue of the face based on a solid structure
formed by the bone of the head. In particular, the zygomatic and malar bone play a role preventing the
falling down of the soft tissues. The higher is the malar and zygomatic bones volume and then develop
out of the face and the greater is the support. Today we can help patients with low zygomatic and
malar bone volume by increasing the volume of this with an autologous bone implants.
Intervene using the platelet poor plasma that remains from the second bio-stimulation (see above) and
joining it to a biomaterial compatible with bone tissue. We use the tricalcium phosphate powder in 30
micron microspheres in sterile 0.5 grams vials. The suspension is prepared by mixing tricalcium
phosphate plasma poor in platelets in a concentration of 20%. Once prepared, the suspension, it is
homogenized by a vortex. Then, the syringe is placed in a water bath to coagulate plasma proteins with
the heat with training of STBA (Support Autologous Biologic Tissue). We obtain a syringe with
autologous proteins mixed with tricalcium phosphate. The product is inject at the periosteum in the
malar-zygome.

7. The patient's face is drawn with two lines: the first from the corner of the nostril to the top of the
tragus and the second from the corner of the mouth outside corner of the eye. In the junction point
we enter with the needle, perpendicular to the tissues, reaching the bone. Continue parallel to the
bone, both at the level zygomatic malar. We inject slowly withdrawing the needle. To end we rub to
homogenize the product on the periosteum.
Within 30-40 days, the fibrotic foreign body answer leads to increased volume treaty bone.
Treatment may be repeated up to aesthetic improvement.
Regeneration of the face adipose tissue
The regeneration of the face adipose tissue follows the outline of a technique already in use: the
lipofilling. This provides the removal of a certain amount of fat from a home device body and the
replanting of the same, treated or not, in the face.
Regeneration does not use the technique of normal fat cells but mainly of fat stem cells. These are
sown in small amounts to stimulate the formation of new adipose tissue (liposowing).
Fat is rich in stem cells. A stem cell is every 50 normal cells (compared to bone marrow that contains 1
for 10000).
Today there is much talk of the use of stem cells present in adipose tissue. Much to develop a machine
(high cost) that can increase and separate the stem cells of the fat.
Professor J. Victor Garcia and Prof. Maurizio Ceccarelli have developed a simple technique to enrich
stem cells in the area of collection for Liposowing. This technique has been presented for the first
time at BioBridge Event of 2008 in Geneva Palais des Nations.
The explanation of why the fat is so rich in stem cells be found in the need to accumulate energy by
adipose tissue in a state of adipocyte hypertrophy.
The adipocyte is able to significantly increase its volume to collect energy in the triglycerides form.
But when its volume is very high (greater than 170% of normal volume) the adipocyte stimulates the
formation of new adipose tissue by activating the differentiation of stem cells in the stroma-vascular
connective tissue.
The liposintetic stimulus leads to adipocyte hypertrophy that, at a certain volume, to stimulate
perivascular stem cell propagation and differentiation.
The stimulus to mitosis and differentiation preadipocytes follows mainly to increasing the rate for
insulin receptor down regulation in the hypertrophic adipocyte and the liberation IGF-1.
The insulin receptor down regulation creates insulin resistance. It follows to a local concentration gain
of insulin and to a preadipocytes stimulation. The volume adipocyte increase active paracrine secretion
of IGF-1 and stimulates the preadipocytes formation.
The liposowing differs therefore from lipofilling mainly because fat stem cells are implanted.
Based on the foregoing, the Liposowing provides:
1. Stimulation of the fat donation area of body with glucose and insulin increase the volume
adipocyte ready. Insulin, which main lipogenetic hormone, stimulates the function of
lipoproteinlipase allowing fatty acids uptake of circulating lipoprotein and the entry of glucose
in the adipocyte. Glucose is the precursor of glycerol phosphate. The latter binds fatty acids
to form triglycerides. It uses an amount corresponding to 1 IU of insulin per kilo of fat to
stimulate.
2. 100 units of insulin are diluted in 250 cc of saline solution or glucose 5%. The resultant solution
with 0.4 U.I. per milliliter.
3. We use a milliliter of this solution together with 200 ml of 5% glucose to stimulate an area of
fat removal of 400 cubic centimeters.
4. Inject 0.5 milliliters per cubic centimeter of fat on the prepared solution with a concentration
of 1 IU insulin per fat stimulate kilo and a concentration of 5% glucose.
5. After 4 hours of the first infiltration if it makes a second adding 1% local anesthetic and
adrenaline.

8. 6. To the bleaching of the tissue we sample with 14 G needle to allow the collection of the stromal
fraction stem cells rich. It's important the collection of stromal-vascular fraction because this
contains the stem cells.
7. For implantation using small cannulas (2.1 mm) or needles cannulas 14 G. The plant is run by
distributing small amounts (like Fischer grains of rice) and to facilitate a smooth distribution
may be done with a special gun.
The technique of Liposowing with fat stem cells, allowing a system that allows to a better engraftment
of fat cells and to a differentiation of stem cells, not only as adipocytes, as well as dermal-epidermal
cells.
By Medical Face Lifting can be restored to the body tissues of the face allowing for a comprehensive
rejuvenation of this.
The Practice
See you now, on a practical level, the Medical Face Lifting.
By Medical Face Lifting we can regenerate:
• Epidermis
• The dermis
• The hypodermis
• Bone
We use the term Full Face Regeneration.
To achieve the above we use the patient's autologous products including:
• Autologous Platelets Derived Growth Factors
• Platelets Rich Plasma
• Plasmatic Autologous Fibrin
• Autologous Fat Stem Cells
• Autologous Biological Tissue Support
We start at time 0, with the dermal regeneration through autologous platelet growth factors.
We take a sample of venous blood to the patient. We need, therefore, a system Vacuette, a butterfly
needle and harvesting tubes. The tubes contain the anticoagulant sodium citrate at 3.8%.
Sodium citrate in the presence of calcium ions, forms calcium citrate. The sequestration of calcium
leads to the impossibility of passing the prothrombin into thrombin and thus prevents blood clotting.
There are kits certified for blood harvesting to be used in the preparation of platelet factors. These
tubes contain a gel cap that allows to separate the majority part of the corpuscular centrifuge in the
lower and plasma and platelets and lymphocytes high. In particular, the spin take under the cap gel red
blood cells, neutrophils, and eosinophils, leaving above the cap gel platelets, lymphocytes and
monocytes.
The centrifugation time recommended in the literature for the platelets separation is 1500-1800 RCF
(relative centrifugal force) for 15-18 minutes. Our practice is sufficient for half the time (750-800
RCF for 7-9 minutes).
To get the speed we consider the radius of the centrifuge. A simple nomogram correlates cm radius
with RCF giving us the speed of our centrifuge program.
After separation by centrifugation, stirring gently the tube for mix the platelet concentration
throughout the volume. Then, we collect the whole plasma.
If we use the kit with the gel tube is easily sucks the whole plasma. In the case of using a plain tubes
(no gel) should be careful do not take with the aspiration the leukocyte portion. Here, in fact, we have
granulocytes that can be released after the activation, proteolytic enzymes, cathepsin,
myeloperoxidase and toxic compounds with oxygen can damage the skin.
During centrifugation of blood, we prepare the skin of our patient using a mask anesthetic. We treat
face, neck, décolleté and hands.

9. We perform now a biostimulation on all called areas with a dermal infiltration of plasma with a needle 4
mm 30 G. It’s important to examine the formation of wheals, sign of an intradermal introduction.
Platelets introduced are activated, degranulate and release PDGF.
It is the calcium that induces platelet degranulation. This can be released by activating endogenous
platelet collagen receptors or by adding plasma exogenous calcium chloride.
The intradermal introduction brings platelets into contact with the dermal collagen and to their
activation and subsequent degranulation. In this case we can use plasma to complete a single syringe
because degranulation occurs only when the platelet is introduced into the dermis.
In the case of exogenous activation, add to plasma calcium chloride 10mEq/10 ml.
The degranulation following the addition of the activator with release of PDGF. In this case, the PDGF
is released into the syringe and, for its short half-life, we must be very quick to introduce it into the
skin. It’s better to start up small quantities (1 ml) of plasma having a good time before the inactivation
of growth factor.
After completing the treatment areas should be thoroughly sterilized and at least thirty minutes, the
patient should not put on skin creams or tricks.
6 hours after the biostimulation with platelet growth factors we make a second dermal biostimulation
with plasma rich in platelets. This is to enhance dermal regeneration.
In fact, the first bio-stimulation induces fibroblasts a process called receptor recruitment. This
means that the first activation attract new receptors on the wall of the fibroblast and the second
induces a greater metabolic response.
The skin to be treated (face, neck, décolleté and hands) is always prepared with a sterilization and
application of a anesthetic mask.
We make the usual sampling with tubes containing anticoagulant (sodium citrate). It is taking 20 ml of
venous blood in two tubes. Better use of the kit certified for this treatment. The blood is centrifuged
respecting the necessary parameters to obtain a proper separation of platelets.
This time he picks up only the lower third of the plasma. The area is rich in platelets. The remaining
plasma is used for bone regeneration.
We introduce the plasma into the dermis directly. In this case, the platelet activation follows the
contact of these with the dermal collagen. This mode allows us to operate with a longer time using a
single syringe with the whole plasma.
We can, however, activate the platelets, prior to the introduction, with calcium chloride. In this case
we must be very quick to use growth factors released prior to their inactivation. It’s better to split
the plasma into 1 ml syringes, activate it and insert it into the skin in few minutes. Repeat the process
until the use of whole plasma.
Again, after treatment, the area must not be contaminated with creams or tricks and must be
sterilized.
The plasma remaining prior use, poor in platelets, is used to prepare the Autologus Biological Tissue
Support that is added to tricalcium phosphate. Take the remaining plasma, pour it in vials containing
0.5 g tricalcium phosphate with a particle size of 30 microns, homogenize the emulsion formed by
shaking with a Vortex. We have the emulsion in a syringe in a boiling water bath. It has the coagulation
of proteins and obtain a filler formed by Autologus Biological Tissue Support (coagulated plasma
proteins) dispersed with tricalcium phosphate 20%.
Now we can use this product to regenerate the zygomatic bone, increasing the volume. The volume
increase zygomatic bone results in a lifting and repositioning of the tissues fell by gravity and aging.
The procedure involves:
1. Draw two lines, one from the outer eye outer corner of the mouth and the outer edge of the nostril
to the upper edge of the tragus;
2. In the point of intersection we enter with the needle perpendicular until it touches the bone;
3. We rotate the syringe and walk pass the needle to the bone;
4. The product is injected radially covering both the malar and zygomatic bone;
5. We massage to distribute the product evenly on the bone.

10. The granules of tricalcium phosphate induce a fibrotic foreign body response which is seen, in 30-40
days, with an increase in bone volume.
After 10 days of the first biostimulation we have the maximum angiogenic response.
The tissue of the face is therefore best placed to facilitate the engraftment of adipose stem cells,
useful for the hypodermis regeneration.
To increase the concentration of adipose stem cells we must induce the proliferation of these in the
area of sampling. Insulin and IGF-1 have this feature and act when the adipocyte volume exceeds
170%.
To stimulate the increase in adipocyte volume it stimulates new formation of triglycerides infiltrating
the area with Ready Insulin and Glucose Solution.
We perform an initial dilution putting 100 IU of Insulin Ready in 250 cc of glucose solution 5%. We get
a concentration of 0.4 IU of insulin per milliliter.
Considering that the amount of insulin normally used is 1 IU per kg, if we want to stimulate an sampling
area of 20 cm for 20 cm to 400 cc of fat, we use 0.4 IU and then 1 ml of solution that we have
prepared.
We prepare the solution to infiltrate the 400 cc of fat, like this:
• Take 1 ml from the solution already prepared;
• Put a milliliter in 200 ml of glucose 5%.
• We use all this new solution (200 ml sol. Glucose 5% + 0.4 IU of insulin) to infiltrate the sampling
area (400 cc of fat). This is equivalent to a volume of 0.5 cc per cubic centimeter of fat.
• Wait four hours to allow the formation of new triglycerides intraadipocyte and the resulting
proliferation of stem cells by insulin and IGF-1.
• We perform a new infiltration preparing a solution of 200 ml of Sol 5% glucose with 0.4 IU insulin
and adding 5 ml of 1% lidocaine with 1:50,000 Adrenalin.
• Expect the sign of complete bleaching of the zone for action of the anesthetic and
vasoconstrictor.
• We perform the aspiration of fat stem cells using 14 G needle mounted on a 5 ml syringe. This is
to collect the stromal-vascular fraction, where are the stem cells, and to avoid injury to these
cells, very sensitive, with excessive intake. The needle with the sharp point, pick up real small
fraction-vascular stromal cores. Stem cells (CD34 +/CD31-) differ into adipoblast (CD34+/CD31+)
that are implanted more easily and, with their propagation give rise to the formation of a new fat.
We can use, for withdrawal, the Lipivage syringe. This has the advantage of allowing the
separation of fat from the anesthetic solution and blood, very easily, but as a contraindication to
its excessive volume (100 ml) and the suction force (too high).
• After removing the fat with stem cells and pending the preparation of the planting area (cleaning,
sterilizing and anesthetic) is put into syringes still the glucose solution with insulin.
• Upon liposowing, emptying the syringes of the solution and inject the fat with stem cells in small
quantities distributed by a micro cannula 2.1 mm in areas where fat tissue is hypotrophic (bubble
Bichat, nose geniene angle bottom of mouth).
• After liposowing antibiotic cover is maintained for three days.
After 30 days of the first biostimulation we have the highest concentration of activated
fibroblasts.
Optimize, then, our dermal regeneration by making a carpet biostimulation with amino acid precursors
of matrix components and a bicarbonate buffer.
We use a Level III Medical Device ready and contains the amino acid precursors of the components of
dermal matrix combined with a bicarbonate buffer capable of maintaining a physiological pH of the
skin. We conduct carpet intradermal infiltration on all areas (face, neck, décolleté and hands) using 3
vials of the product. Amino acids in the Medical Device allow activated fibroblast to regenerate the
skin dermis. The proline is used as a precursor of collagen, glucosamine as a precursor of hyaluronic
acid and lysine as a precursor of desmosina, flexible hinge elastin. Bicarbonate buffer enhances the

11. status of the dermal matrix keeping its flow through maintaining a pH of 7.4. This also allows to avoid
a new formation of fibrous collagen (type I) characteristic of inflammation (acidic pH).
The skin, before treatment, is always prepared with a anesthetic mask.
The Medical Device is introduced with small intradermal wheals, carpet in all areas, using a needle 30G
4 mm.
To the dermal regeneration we match a particular activation of fibroblasts in the district of wrinkles
to reduce these imperfections. We inject Autologus Plasma Fibrin to create a protein mesh under the
wrinkle. This mesh is colonized from the fibroblasts that reconstruct the dermis preferentially in this
district. We get a filler effect that reduces wrinkles and depressions.
The fibrin is formed by the action of thrombin on fibrinogen. The newly formed fibrin polymer is then
stabilized by factor XIII. The fibrin forms the fibrin clot that prevents blood loss from injury.
Rebuilt the endothelial wall, its cells produce a particular activator that converts circulating
plasminogen to plasmin. The plamina degrades the fibrin clot and recanalise the vessel.
We make a withdrawal of venous blood without anticoagulants.
We centrifuge to separate the plasma from the cells. Meanwhile, the prothrombin is converted into
thrombin and active fibrin clot formation. primum movens of the same lysis.
In addition, to increasing the time duration of the clot under the wrinkle, add a drop of tranexamic
acid prevents the conversion of plasminogen to plasmin.
We inject as a filler the Autologus Plasma Fibrin to activate a preferred reconstruction of the dermis
in the district of our interest.
We complete the facial tissues regeneration with the epidermal treatment using Plasma Rich In
Platelets.
The skin should be prepared prior to application with a mild skin abrasion that removes the corneous
layer and with a mandelic acid peel to open desmosomes ties to allow an epidermis splitting useful to
epidermal penetration of active ingredients.
The Protocol forsee:
1. Abrasion of the corneous layer
2. Peeling with mandelic acid
3. Topical application of plasma rich in platelets
4. Occlusion with a collagen thin layer to improve the penetration of platelet growth factors.
• We make a blood vein withdrawn using tubes with sodium citrate at 3.8%.
• Centrifuge and remove the lower third of the plasma (PRP).
• Platelet Rich Plasma sprayed on the skin abraded and peeling cracking.
• Finally, apply a thin layer of collagen to occlude the skin and facilitate penetration of active
ingredients.
The upper two thirds of the plasma (PPP) is used for a deep correction of wrinkles (in case of
insufficient regeneration of the dermis with Autologus Plasma Fibrin), mixed with Tricalcium
Phosphate.
The operating scheme is the same as bone regeneration, but to obtain the emulsion we use a smaller
amount of tricalcium phosphate (0.1 gr.) to give a final concentration of 4%.
The emulsion is coagulated with boiling water bath and finally obtain 2 ml of Biological Autologus
Tissue Support with 4% of tricalcium phosphate. This coagulated emulsion, effect filler, is used for
a final wrinkles correction like using a synthetic filler.
The introduction is made in the deep dermis, in small amounts and with a long massage after filling. The
presence of tricalcium phosphate granules induces a fibrotic response that is revealed as a cosmetic
correction.
This is the practical protocol of Medical Face Lift in a manner best described by the term Full Face
Regeneration.

12. In summary:
1. At time 0 we make an initial dermal regeneration using Autologus Platelet Growth Factors;
2. At 6 hours after time 0 we carry out a further dermal regeneration using Plasma Rich In Platelets;
3. Simultaneously (6 hours from time 0) we make a bone regeneration using Autologus Biological Tissue
Support emulsion with tricalcium phosphate 20%;
4. At 10 days time from 0 (maximum angiogenesis), we make a hypodermis regeneration using
Autologus Adipose Stem Cells;
5. In 30 days time from 0 (maximum concentration of activated fibroblasts) a further dermal
regeneration is carried out using aminoacids precursors of the dermal matrix components and a
bicarbonate buffer;
6. Simultaneously we activate the dermal reconstruction in wrinkles and depressions with an
Autologous Plasma Fibrin infiltration;
7. Furthermore, we make a epidermal regeneration using Platelet Rich Plasma for topical
administration on abraded and cracked with peeling skin;
8. Also we correct the unsightly residue of wrinkles and depressions with Autologous Biological Tissue
Support emulsified with tricalcium phosphate to 4%.
All this allows for a face rejuvenation of our patients without surgery using and regenerating the
biological state of tissues.
Bibliography
Ceccarelli M., García J. V., Il Medical Face Lifting: Rigenerazione dei Tessuti del Volto, The
Physiological Medical Letter Vol. I, N° 1, Gennaio 2010