Autologous tissue regeneration with the Outin preparation involves the use of biologically activated patient plasma enriched with DNA nucleotides, which represents a "reservoir" of growth factors released from blood platelets - platelets. The method is used for:
- 1) skin renewal and fight against stress
- 2) repair of cellular damage from UV rays
- 3) treatment of scars and skin lesions (post acne)
- 4) photoaging
- 5) skin hypoxia
- 6) skin before and after menopause
- 7) anti-wrinkle treatment
- 8) optimized shine and brightness of the skin
PRP-"VAMPIRE FACELIFT" - DEFINITION
PRP-"platelet-rich plasma" or plasma rich in platelets is the patient's own blood with an increased concentration of platelets, which upon activation release growth factors (proteins), that is, it is a preparation obtained by centrifuging whole blood in order to increase the concentration of platelets.
Blood: number of platelets 150-350 x 10 9 in 1 mm3 of blood + anticoagulant.
PRP: soft spin: 1000 revolutions for 5-6 minutes.
Prf: 1300-2700 revolutions over 8-12 minutes.
PRP scientific works
- a method developed in the 1970s
- first time of use in Italy in 1987 (open heart surgery)
- surge in popularity in the mid-1990s
- the number of peer-reviewed publications has increased sharply since 2008.
- a total of more than 5,700 scientific papers
The most commonly used blood derivatives in clinical use:
- platelet rich plasma (prp): platelet rich plasma
- platelet-rich growth factors (prgf)
- platelet-rich fibrin (prf)
- plasma enriched with platelets and leukocytes (l-prp)
- fibrin enriched with platelets and leukocytes (l-prf), etc.
Platelets - history of discovery
Throughout history, before and after the scientific revolution, until today, scientists and naturalists have studied platelets, which allows us today to know a lot about platelet structure, function and the life of platelets in general. They were discovered by the French doctor Alfred Donne in 1842. Giulio Bizzozera, in 1882 established the fundamental knowledge about platelets, which were confirmed after him. Studying animal veins microscopically, he discovered that the shape of platelets is discoid, and that after an injury to a blood vessel in the circulation, the process of adhesion and aggregation begins: a role in the process of hemostasis. He was the first to define platelets as central cells of the clotting system. Before the discovery of platelets, Bizzozero also identified megakaryocytes in the bone marrow, which are also platelet precursors.
About 40 years ago, platelets were considered primarily cells responsible for hemostasis (stopping bleeding).
Thrombocytes are blood platelets, i.e. small, colorless corpuscles without a nucleus. There are 150-400 x109/1 mm3 of blood in a liter of blood. They are formed in the bone marrow from parts of megakaryocytes (cells with a large nucleus). They decompose quickly: their lifespan is from 3 to 7 days.
Photo: from left to right: erythrocyte, platelet, leukocyte
PRP concentrates platelets about five times.
What is special about platelets?
Platelets contain alpha granules that contain a large amount of growth factors and dense corpuscles. Alpha granules are released by the aggregation and adhesion of platelets at the site of injury or inflammation.
Abundance of granules in platelets
The most interesting property of platelets is the abundance of granules that contain a large number of biomolecules, among which is serotonin. We distinguish 3 types of granules: αalpha-granules, dense granules that are a powerful storage of serotonin (eng. Dens granules) and lysosomes. Since the 90s of the last century, scientific research and technology have shown that platelet GRANULES contain abundance of growth factors. Growth factors play a key role in the HEALING AND TISSUE REGENERATION PROCESS. Growth factors stimulate the migration and multiplication of stem cells.
The most interesting property of platelets is the abundance of granules that contain a large number of biomolecules, among which is serotonin. We distinguish 3 types of granules: αalpha-granules, dense granules that are a powerful storage of serotonin (eng. Dens granules) and lysosomes. Since the 90s of the last century, scientific research and technology have shown that platelet GRANULES contain abundance of growth factors. Growth factors play a key role in the HEALING AND TISSUE REGENERATION PROCESS. Growth factors stimulate the migration and multiplication of stem cells.
Schematic representation of the platelet structure
Growth factors
- platelet-derived growth factor (PDGF)
- fibroblast growth factor (FGF)
- transforming growth factor beta (TGF-β)
- insulin-like growth factor 1 (IGF-1)
- insulin-like growth factor 2 (IGF-2)
- vascular endothelial growth factor (VEGF)
- epidermal growth factor (EGF)
- keratinocyte growth factor (KGF, FGF7)
- connective tissue growth factor (CTGF, CCN2)
- interleukin 8 (IL-8)
PDGF (platelet-derived growth factor) - a group of polypeptides that stimulate protein synthesis - especially collagen and angiogenesis
TGF beta, EGF, VEGF they stimulate angiogenesis and synthesis of collagen, cell matrix, fibronectin
PDEGF (platelet-derived epidermal growth factor) stimulates the proliferation of keratinocytes and fibroblasts
PDAF (platelet-derived angiogenesis factor) stimulates angiogenesis
IGF-1 stimulates the synthesis of cell matrix elements and the mitotic activity of stem cells
PF-4 (platelet factor 4) attracts fibroblasts
Activation of platelets and release of growth factors is what makes this therapy so widely applicable and interesting!
What is the role of growth factors?
CHEMOTAXY
- cell migration in response to a chemical stimulus
- stem cells are "attracted" by growth factors and migrate towards the site of application
MULTIPLICATION OF CELLS
- significant increase in the mitotic activity of cells
Effect of growth factors on tissues
Immediate (within 5 minutes)
- stimulation of other messengers
- inflammatory response
Early (30 minutes to 4 hours)
- increase in M-RNA expression, induction of protein synthesis
- chemotaxis (attraction of cells to the application region)
Late (4-24 hours)
- increase in mitotic activity of fibroblasts
Deferred (up to 3 months)
- tissue remodeling
The overall effect of platelet-derived growth factors on tissues
- they interfere with each other in a complex manner with different effects on cells
- they cause a series of biochemical reactions and cell modifications, leading to a rapid and long-term increase in the number and activity of fibroblasts
- they modulate cellular activity that leads to the division and maturation of stem cells, which is not fully explained
Indications for application
- deep skin rejuvenation
- wrinkle reduction
- reducing the visibility of scars
- acne (scars)
- stretch marks
- postpartum skin laxity
- alopecia
- treatment of cellulite
Expected results
- improvement of skin texture and tone after 3 weeks of application
- collagen synthesis and remodeling within 3 months of application
Preparation of platelet-rich plasma
Blood extraction
About 10-15 ml of blood is taken from the client into test tubes filled with anticoagulant (Na citrate).
Centrifugation
The blood is centrifuged at 1000 revolutions or 300g for 6 minutes, which separates the blood into 3 compartments: the upper and middle "buffycoat" rich in platelets and leukocytes, and the lower one rich in erythrocytes.
Separation of platelet-rich plasma
Platelet activation
Platelet activation is carried out with an activator containing calcium chloride, adenine, cytosine and thiamine
Application
Result of blood conditioning
On average, blood contains 2x 105 platelets per microliter, and centrifugation produces plasma rich in platelets, which clots up to 5 times higher concentration of platelets
Application methods
Injection techniques (SUPERFICIAL AND DEEP)
Papules: Surface intradermal technique, the application site is the junction of the epidermis and the dermis
Nappage: The technique was designed by Dalloz Bourguignon, the application is made with a series of injections at a depth of 2 to 4 mm, maintaining constant pressure in the syringe that is inserted into the skin at an angle of 30 to 60 degrees
Point by point: Deep intradermal and hypodermal technique at a depth of 4 mm and more
Microinjection techniques ("derma roller" and "derma pen")
Alpha granules and growth factors
Alpha granules release growth factors approx. 10 minutes after "activation" of platelets. The viability of platelets and the release of growth factors into the tissue continues until the end of the life of the platelet. In addition, platelets stimulate the influx of macrophages and stem cells, which additionally contribute to the process of tissue restoration and regeneration through the removal of degenerated and necrotic tissue, and stimulate tissue regeneration.
Growth factors stimulate regenerative processes: chemotaxis, proliferation, differentiation and angiogenesis.
PRF
PRP concentrates platelets about five times.
PRF represents a newer generation of platelet concentrates.
The special texture of PRF allows clinical use in amorphous and membrane form. The membrane form is used to cover and protect large tissue grafts. It stimulates osteogenesis and angiogenesis.
The first promotion of PRP
Dr. M. Ferrari 1987:
Open heart surgery and autologous blood component transfusion to avoid the use of homologous blood transfusion after open heart surgery.
Application from: orthopedics, sports medicine, dentistry, otolaryngology, neurosurgery, ophthalmology, urology, wound healing, aesthetic medicine, cardiothoracic and maxillofacial surgery.
PRP is an autologous product, the possibility of an immunogenic reaction and transmission of infectious diseases such as HIV and hepatitis is eliminated.
PRP – well-being
- in application and research since 1970.
- In 1990, the beginning of application in plastic surgery and facial and jaw surgery
- the healing potential of muscles and tendons introduced this therapy to orthopedics in the early 2000s.
- PRP began to be used as a treatment for tendinopathies in the early 1990s
Recently documented beneficial effects of PRP on chronic non-healing injuries of tendons, ligaments, muscles, bones (osteoarthrosis), intervertebral disc degeneration, plantar fasciopathy and cartilage degeneration as well as in the care of muscle trauma and ischemic nerve damage.
PRP - effects
- tgf-β1 and tgf-β2 are involved in the regeneration of connective tissue and bone
- they stimulate fibroblast chemotaxis and collagen production
- inhibit the breakdown of collagen by reducing the activity of proteases and increasing the activity of their inhibitors
- in vitro and in vivo studies have shown that TGF increases the proliferation of mesenchymal stem cells and osteoblasts
The growth factor found in plasma rich in transforming growth factor-β (TGF-β) stimulates:
- proliferation of mesenchymal cells
- regulates endothelial, fibroblastic and osteoblastic mitogenesis
- regulates collagen synthesis and collagenase secretion
- regulates the mitogenic effects of other growth factors
- stimulates endothelial chemotaxis and angiogenesis
- inhibits the proliferation of macrophages and lymphocytes
- fibroblast growth factor (fgf) promotes the growth and differentiation of chondrocytes and osteoblasts
- mitogenetics for mesenchymal cells, chondrocytes and osteoblasts
- platelet-derived growth factor aib (pdgf) for mesenchymal cells and osteoblasts
- stimulates chemotaxis and mitogenesis in fibrillar, glial or smooth muscle cells
- regulates collagenase secretion and collagen synthesis
- stimulates macrophage and neutrophil chemotaxis
- epidermal growth factor (egf) promotes endothelial chemotaxis or angiogenesis
- regulates the secretion of collagenase
- stimulates epithelial or mesenchymal mitogenesis
- vascular endothelial growth factor (vegf) increases angiogenesis and permeability
- stimulates mitogenesis for endothelial cells
- connective tissue growth factor (ctgf) promotes angiogenesis
- cartilage regeneration
- fibrosis and adhesion of platelets
- insulin as a growth factor (ilGF 1 and 2) for fibroblasts and stimulates protein synthesis
PRP has received attention in the popular media as a result of its use in the treatment of sports injuries in professional athletes:
Tiger Woods
Born as Eldrick Tont Woods – American golfer. He is considered one of the greatest golfers of all time.
Raphael Nadal
Rafael "Rafa" Nadal Parera, professional Spanish tennis player. Nadal has won fourteen Grand Slam titles, the 2008 Olympic gold medal and 28 Masters 1000 titles and was a member of the Spanish Davis Cup team that won the titles in 2004, 2008, 2009 and 2011.
Kim Kardashian
"Vampire facelift"
Safety of use
Hypothesis: overexpression of growth factors and their receptors associated with the formation of tumors and tissue dysplasia.
Growth factors regulate various cellular processes, such as mitogenesis, chemotaxis, cell differentiation and metabolism. Neoplastic growth requires higher continuous doses of growth factors over a longer period of time. When applying PRP therapy, extracellular growth factors are broken down within 7-10 days.
Advantages of using: Autologous PRP - no risk of cross-reactivity - immune reaction or disease transmission.
Advantages of PRP compared to other methods of rejuvenation
- it uses its own platelets, so the risk of allergic reactions is minimal
- enhances collagen synthesis in response to the secretion of growth factors
- enhances the synthesis of hyaluronic acid and the amount and density of the extracellular matrix
- ideal for those clients who do not want synthetic fillers
- swelling, bruising and bruising are significantly less pronounced than with other methods
- can be combined with other methods
- it is equally effective in men and women
Combining with other rejuvenation methods
PRP can be combined with:
Filers – Fillers lead to an immediate increase in volume, and injecting PRP above the filler improves the texture and tone of the skin and extends the effectiveness of the filler for 3 to 6 months
Laser therapy – Application of PRP after laser therapy significantly accelerates recovery and gives better final treatment results
Peelings – Accelerates healing after the application of medium-deep peels and improves the tone and texture of the skin
Absolute contraindications
- platelet dysfunction
- thrombocytopenia
- hemodynamic instability
- septicemia
- active infection in the application area
- client's reluctance to accept risks
Relative contraindications
- NSAID use 48 hours before the procedure
- application of corticosteroids in the application area one month ago
- systemic use of corticosteroids 2 weeks ago
- fever back 14 days
- neoplasms, especially of the hematopoietic system
- HGB 10 g/dl
- platelet concentration 105/ul
- chemotherapy
- hypofibrinogenemia
- anticoagulant therapy
- chronic liver diseases
- systemic lupus erythematosus
- porphyria
- allergies
- abuse of nicotine products and alcohol
Possible side effects
- infection
- discoloration
- bruises
- soreness in the application area
Practiced preparation and application is the key to success
As with other therapies, thorough training and experience are the most important prerequisites for an optimal treatment result!