Inna SHARYPOVA, M.D., Ph.D. Andrey ALENICHEV, M.D., Ph.D.

Dermatologists, Kristel Medical, Milan


Today, dermatologists and cosmetologists have a wide arsenal of tools at their disposal to treat age-related skin changes, including various medical equipment, injection methods and cosmeceuticals. Age-related skin changes are a task that cannot be solved using state-of-the-art technologies alone.

  • This is why methods that combine different mechanisms acting on skin cells and the extracellular dermal matrix are highly relevant.
  • The main aim of any combination is to improve the correction effect and reduce adverse reactions.
  • To achieve this, it is necessary to improve the condition of the skin in many ways: from surface characteristics to profound changes in cellular function and tissue structure.

We can achieve these goals by reinforcing treatment protocols with methods that support repair processes by improving microcirculation and tissue regeneration.


Today, a number of physical methods can be used to correct the signs of skin ageing: laser radiation, focused ultrasound, radiofrequency electric currents, broadband light, etc.

  • The medical devices used in cosmetology induce controlled micro-traumas that disrupt the physiological balance of skin tissue and trigger the restructuring processes that follow.
  • For example, the mechanisms of fractional radiofrequency (RF) microneedling are mainly based on the thermal effect and aseptic inflammation at the site of controlled microtrauma.
  • Their appearance is directly linked to the frequency of the alternating current passing through the skin - the lower the RF, the more pronounced the effect.

Compared with non-ablative fractional photothermolysis, fractional radiofrequency microneedling delivers energy to deeper layers of the skin.


  • Under the influence of the physical factors underlying methods based on fractional radiofrequency Microneedling, a greater number of cells in the epidermis, dermis and subcutaneous tissue move into an active state. This leads to structural and functional improvements.
  • The desired effect of the device's use is only possible when there is an optimal balance between the stress factor, represented by the device's action, and the tissue's ability to adapt...
  • To enhance and optimize the skin's ability to regenerate and replenish itself, biological injection methods are used, in particular autologous platelet-rich plasma (PRP) therapy.


The effectiveness of PRP therapy relies on the triggering of reparative and trophic processes activated by various paracrine factors present in platelets.

  • PRP has a strong healing and regenerative potential, as it is a concentrate of the main bioactive substances involved in the various stages of the healing process.
  • The effect of PRP is due to degranulation of platelet alpha granules, which contain glycoproteins and growth factors including platelet growth factor, transforming growth factor beta, fibroblast growth factor, vascular endothelial growth factor and epidermal growth factor.
  • In particular, the biological "cocktail" secreted by platelets has the ideal physiological proportion of growth factors. On contact with a surface different in structure from the vascular endothelium, and under the action of thrombin, platelets become activated, transformed and produce pseudopods that bind them to the fibrin network.
  • Under the action of growth factors, dermal fibroblasts synthesize collagen, elastin and a non-fibrous matrix that strengthen and lift the skin.
  • In addition, new capillaries form (angiogenesis) and the epidermis renews itself. To achieve a significant clinical effect, it is essential to have a sufficient number of viable platelets which, after being activated in the tissue by the patient's own thrombin, release growth factors and other bioactive substances into the surrounding intercellular space.

Essential conditions must be met for PRP preparations:

  • Platelet concentration must be 1.5 to 2.5 times higher than in blood. In particular, concentrations 2.5 to 3.0 times higher than physiological values lead to mutual inhibition of platelets and a marked reduction in the release of growth factors;
  • the absence of heparin, which blocks the ability of platelets to aggregate on fibrin, thus blocking the physiological process of activation and release of growth factors;
  • cells must not be mechanically destroyed during preparation, as growth factors only become biologically active when preformed by crossing intact, functionally active platelet membranes;
  • a minimum number of granulocytes, which have pro-inflammatory activity;
  • absence of red blood cells ;
  • presence of monocytes, which enhance the efficacy of the preparation by reinforcing non-specific immunity.

Injectable hyaluronic acid (HA) preparations are well known to cosmetologists.

In the skin, HA molecules over 500 kDa create the conditions for cell migration and proliferation, and maintain homeostatic equilibrium. Today, we also have a combined approach for intradermal injections: Cellular MatrixTM.

  • RegenLab's patented medical device contains the patient's own autologous PRP and a certain form of hyaluronic acid.
  • The non-cross-linked hyaluronic acid contained in the tube acts as a scaffold-like structural element, while PRP has a restorative and regenerative effect. The presence of hyaluronic acid exactly at the moment of fibrin polymerization enables the formation of three-dimensional structures in which platelets are dispersed.
  • These 3D structures are colonized by fibroblasts, enabling increased synthesis of extracellular matrix components and organized dermal reconstruction.
  • When the preparation containing the patient's autologous PRP and HA is injected into the target tissues, a biologically enriched network is formed, a matrix that facilitates cell migration and proliferation, improves tissue hydrodynamics and ensures the distribution of trace elements, metabolites and hormones.


For the treatment of signs of skin ageing, particularly in the face and neck areas, we use a protocol that combines PRP injections and radiofrequency microneedling.

For PRP therapy, we apply two RegenLab medical devices to each treatment:

  1. The RegenLab kit Plus device with additional activation of PRP with autologous thrombin serum (ATS) to form an autologous PRP gel used to fill wrinkles and improve facial contours.
  2. Device Cellular MatrixTM (CM), which combines PRP and 2 ml of non-cross-linked hyaluronic acid (HA) with 2 % in a single tube. It enables a biologically identical material to be prepared in a single step for immediate use in a completely closed sterile system (Figure 1).

The use of CM leads to a physiologically-determined activation of regeneration processes under the action of platelet growth factors and interleukins, while the hyaluronic acid contained in the preparation optimizes tissue hydration, helping to fill in wrinkles and folds.

Figure 1. The stages cell matrix preparation.

Figure 2. Injection diagram.

The overall strategy of PRP injections for the aesthetic correction of age-related changes is specifically aimed at increasing dermal elasticity and turgor. It is advisable to inject the preparation using the linear "filler-like" technique with 30-32G needles.

  • PRP-HA is administered intradermally using the retrograde linear technique for facial soft tissue lifting.
  • It should be noted that in certain areas of the skin, the bolus technique is preferable (Figure 2).
  • Radiofrequency therapy was performed using a Pologen RF device (from Lumenis) designed for minimally invasive fractionated bipolar RF therapy.
  • The RF device's energy has a direct effect (heating) on the epidermis and dermis at a depth of 0.6 to 1.5 mm.
  • Non-insulated microelectrodes deliver lossless RF energy directly to the selected zone, where water evaporates from the microthermal zone.

This stimulates the physiological healing process, as well as collagen and elastin synthesis.

Figure 3. Effect of RF microneedles on tissue.

For skin rejuvenation of the face and body, the treatment consisted of three treatment cycles, performed at one-month intervals according to the following scheme: the RF micro-needling facelift treatment was performed, followed by an injection of RegenLab PRP and cellular matrix one week later.

After a three-week break, this cycle was repeated.

The recommended monthly intervals should be observed between device-based and injection-based treatments.

It is important to note that PRP and cellular matrix injections were performed shortly after radiofrequency stimulation of the dermis.

This contributed to rapid skin restoration after treatment-induced skin trauma, as well as improved neo-collagenogenesis and restored microcirculation.

" Inclusion criteria

Gender - female and male, age - 40-70 years, informed consent to be included in the study, no hypersensitivity to HA and no contraindications to radiofrequency therapy.

" Exclusion criteria

Active skin diseases, acute herpes and other skin infections, exacerbation of general infectious diseases or chronic somatic diseases, mental illness, use of antidepressants or drugs affecting the central nervous system, epilepsy, cancer.

We observed 30 patients with signs of facial skin ageing, aged between 40 and 70, who underwent combination therapy in accordance with the developed protocol.

To assess stratum corneum hydration before and after treatment, corneometry was performed using the CM 825 Corneometer.

Transepidermal water loss (TEWL) was assessed before and after treatment using the Tewameter TM300 vaporimeter.
  • The effectiveness of aesthetic correction of facial skin was assessed using visual analog scales, photographs and a quality-of-life study.
  • Results were compared in two age groups: under 49 (n =1 5) and over 50 (n = 15).


Comparative evaluation of skin functional parameters (hydration and TEWL) showed promising results for combined therapy with cellular matrix and radiofrequency lifting (Table 1).

  • In fact, the hydration of the stratum corneum after the combined therapy was 1.7 times higher in patients under 49 and 2.1 times higher in patients over 50, while TEWL decreased 2.8 times in the under 49 group and 2.2 times in the over 50 group.

Matrix cellular + RF


Under 49


More than 50 years


before treatment

after treatment


before treatment

after treatment



51,1 ± 2,8

23,7 ± 1,6

85,8 ± 3,6

8,5 ± 1,9

P < 0,001

P < 0,01

38,7 ± 3,4

28,1 ± 2,6

79,9 ± 4,2

12,9 ± 1,7

Table 1.    Skin hydration and loss of skin elasticity with RegenLab's fractional radiofrequency microneedling, cell matrix and PRP combination therapy.
Figure 4. Reduction in clinical signs of skin chrono-aging after treatment with the combined method, according to the VAS index (qualitative characteristics of the skin and severity of wrinkles) in patients under 49.
Figure 5. Reduction in clinical signs of skin chrono-aging after treatment with the combined method, according to the VAS index (qualitative characteristics of the skin and severity of wrinkles) in patients over 50.

Assessment using visual analog scales showed a reduction in the main signs of chrono-aging in both groups (Figure 4 and Figure 5).

  • Improved skin quality and reduced severity of mimic and gravitational wrinkles contributed to an overall improvement in patients' quality of life.
  • On average, the DLQI improved by 79.9 % in patients under 49: from 14.5 ± 0.7 to 2.9 ± 0.8 (p < 0.01) (Figure 5), and 75.4 % in patients over 50: from 16.2 ± 0.9 to 4.0 ± 0.5 points (p<0.01) (Figure 6).
  • Another indicator was evaluated in the study - "satisfaction" (efficacy, comfort and safety of treatment); it reflects the patient's opinion based on comparison of expected and received effects in relation to other methods used for similar indications. The overall patient assessment was as follows: 90 % of patients, regardless of age, rated the efficacy and comfort of the combined method as good/very good (Figure 7).
  • Dermatoscopic and photographic data also confirmed the clinical efficacy of the combined therapeutic protocol.

Figure 6. Mean Dermatological Quality of Life Index (DLQI) scores after treatment with the combined method to correct signs of aging of facial skin (y-axis: median values, at p < 0.01 according to the Mann-Whitney criterion).

Figure 7. Data from the patient questionnaire on the efficacy/comfort/safety of the combined method for correcting signs of ageing of facial skin.

Clinical examples of the use of the combined method (radiofrequency facelift and cellular matrix) to correct age-related skin changes.

Before and after combined treatment, patient E. 44 years old

Before and after combined treatment, patient E. 53 years old


The protocol we have developed for combined therapy using bipolar RF micro-needling facelift and injection of RegenLabTM PRP and Cellular MatrixTM is a highly effective method for correcting age-related skin changes, resulting in dermal volumization and skin renewal.

  • We recommend that this treatment strategy be widely used in clinical practice.
  • In addition, PRP can be combined with other equipment used in cosmetology, such as laser, light and ultrasound.
  • Cellular MatrixTM increases the skin's ability to adapt and repair itself.

Significant rejuvenation effects in most patients last up to 1.5-2 years.


  1. Franco W, Kotare A, Ronan Sj and Hyperthermic injury to adipocyte cells by selective heating of subcutaneous fat with a novel radiofrequency device: feasibility studies. Lasers Surg Med. 2010. - С. 361-70.
  1. Sadick NS, Makino Selective electro-thermolysis in aesthetic medicine: a review. Lasers Surg Med. 2004.- C.91-7.
  2. Belenky I, Margulis A, Elman M, Bar-Yosef U, Paun Exploring channeling optimized radiofrequency energy: a review of radiofrequency history and applications in esthetic fields. Adv Ther. 2012 Mar; 29(3): 249-66.
  3. Carruthers Radiofrequency resurfacing: technique and clinical review. Facial Plast Surg Clin N Am 2001. - C. 311-9.
  4. Weiner SF. Radiofrequency Microneedling: Overview of Technology, Advantages, Differences in Devices, Studies, and Indications. Facial Plast Surg Clin North Am. 2019 Aug; 27(3): 291-303.
  5. Friedman Dj, Gilead The use of hybrid radiofrequency device for the treatment of rhytides and lax skin. Dermatol Surg 2007. - 543-51.
  6. Redaelli A, Romano D, Marciano A. Face and neck revitalization with platelet-rich plasma (PRP): clinical outcome in a series of 23 consecutively treated patients. Drugs Dermatol. - 2010; 9:466-72.
  7. Rappl M. et al. Effect of platelet-rich plasma gel in a physiologically relevant platelet concentration on wounds in persons with spinal cord injury. Int Wound J 2011; 8:18.7-195.
  8. Smith J. D., Melhem M. E. et al. Improved growth factor directed vascularization into fibrin constructs through inclusion of additional extracellular molecules. Microvasc Res. 2007. 73(2): 84-94.
  9. al, Effects of PlateletRich Plasma on Intestinal Anastomotic Healing in Rats: PRP Concentration is a Key Factor. J Surg Res. 2012 Apr; 173 (2):258-66.
  10. Turzi, PRP Standardization and Cell Therapy.. Biobridge Foundation, Lausagne 2018.
  11. Girish KS, Kemparaju The magic glue hyaluronan and its eraser hyaluronidase: a biological overview. Life Sci. 2007 May 1 80(21):1921-43. Epub 2007 Mar 6.
  12. Stern R, Maibach HI. Hyaluronan in skin: aspects of aging and its pharmacologic modulation. Clin Dermatol. 2008 Mar- Apr; 26(2):106-22. doi: 10.1016/j.clindermatol.2007.09.013.
  13. Oh JH, Kim YK, Jung JY, Shin JE, Kim KH, Cho KH, Eun HC, Chung Intrinsic aging- and photoaging-dependent level changes of glycosaminoglycans and their correlation with water content in human skin. J Dermatol Sci. 2011 Jun; 62(3):192-201. doi: 10.1016/j.jdermsci.2011.02.007. Epub 2011 Mar 2.

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