Investigation: Multidisciplinary clinical research
REGENPRP®, STANDARDIZED AUTOLOGOUS PLATELET CONCENTRATE.
10 YEARS OF RESEARCH AND PRACTICE.
Pr Barbara HERSANT MD, PhD.
Department of Plastic, Reconstructive and Maxillofacial Surgery, Hôpital Henri Mondor, Créteil, France
Summary
Platelet Rich Plasma (PRP) has emerged as an innovative therapeutic modality in a variety of medical fields, offering promising prospects for wound healing, facial rejuvenation and even genital rejuvenation.
Our clinical research is resolutely committed to studying these applications, seeking to determine the efficacy and safety of RegenPRP® (standardized PRP prepared with technology from RegenLab SA, Switzerland) in these varied clinical settings.
This technology makes it possible to obtain liquid RegenPRP, in the form of autologous fibrin glue or combined with hyaluronic acid (Cellular Matrix technology).®) to meet different therapeutic needs.
In the field of wound healingRegenPRP is attracting growing interest due to its regenerative properties and potential to accelerate the healing process. Our research aims to deepen our understanding of how RegenPRP can optimize the healing of post-operative and traumatic wounds, improving clinical outcomes for patients.
Facial rejuvenation is one area where RegenPRP has become an increasingly popular option. By harnessing the cell regeneration capabilities of RegenPRP, our study examines how this therapy can be used safely and effectively to alleviate the signs of skin aging, offering a non-surgical alternative to traditional facial rejuvenation techniques.
Our groundbreaking research also explores the possibilities of genital rejuvenation using RegenPRP. This innovative approach aims to restore the function and appearance of the genitalia, offering a new means of treatment for patients suffering from sexual dysfunction and aesthetic problems in this area.
It should be noted that all our studies are conducted in compliance with the strictest ethical and regulatory standards. Approved by the French medical agency and supervised by an independent ethics committee, our prospective studies guarantee scientific rigor and participant safety.
Our clinical research on RegenPRP is opening up exciting new perspectives in wound healing, facial rejuvenation and genital rejuvenation by combining a rigorous scientific approach with a commitment to medical innovation. We aspire to improve clinical outcomes and patient quality of life with this promising therapy.
Key words: RegenPRP®Cellular Matrix®RegenPRP association® and hyaluronic acid, plastic surgery, skin grafting, vulvovaginal atrophy, keloid scars, mesenchymal stromal cells.
Figure 1. A. RegenBCT® (10 mL) and B. A-CP kit® 20mL): CE-certified medical devices.
REGENKIT-BCT® AND A-CP-KIT® ARE TWO INNOVATIVE MEDICAL DEVICES FROM REGENLAB FOR THE PREPARATION OF REGENPRP, A STANDARDIZED AUTOLOGOUS PRP.
RegenBCT tubes®available in 10 mL (Table 1) and A-CP®available in 20 mL, offer an efficient solution for the preparation of RegenPRP, a PRP with a standardized composition (Figure 1).
These cutting-edge products reflect RegenLab's ongoing commitment to excellence in healthcare and medical research.
RegenPRP is a regenerative therapy that uses the healing properties of growth factors contained in platelets and blood plasma to stimulate wound healing and tissue regeneration.
Growth factors, such as EGF (epidermal growth factor), FGF (fibroblast growth factor), PDGF (platelet-derived growth factor), TGF-β (transforming growth factor beta), TGF-α (transforming growth factor alpha), IGF (insulin-like growth factor) and VEGF (vascular endothelial growth factor), are bioactive molecules that play a crucial role in the healing and tissue regeneration process.
When RegenPRP platelets are activated by contact with extracellular matrix proteins such as collagen, they release these growth factors, promoting angiogenesis (new blood vessel formation), cell migration, proliferation and differentiation, as well as the synthesis of new extracellular matrix.
This cascade of reactions helps accelerate the healing process and regenerate damaged tissue.
Regen BCT® | Inserts | Red blood cells | White blood cells | Mononuclear cells | Lymphocytes | Monocytes | Granulocytes |
Recovery rates Concentration | ≥ 80%
1,6X | < 0.3% HCT 0.2% 0,007X | 10-13%
0,2X | 20-30%
0,5X | 25-35%
0,5X | ~ 10%
0,2% | 3,5%
0,06X |
Table 1. RegenPRP cell composition prepared with Regen BCT tubes®. HCT: hematocrit.
Figure 2. Simple, effective preparation of platelet-enriched autologous fibrin sealant. RegenPRP in the 10 mL syringe is combined with activated thrombin serum (prepared from patient blood using a RegenATS® tube) and calcium gluconate 10 % in the second syringe using the RegenSpray Applicator.
FIBRIN SEALANT ENRICHED WITH AUTOLOGOUS PLATELETS FOR SKIN GRAFTING
The tubes manufactured by RegenLab are vacuum tubes containing a sodium citrate anticoagulant solution and a chemically inert separator gel. A small quantity of the patient's blood, 10 or 20 mL depending on the device, is drawn directly into the tube. This blood is then centrifuged to separate the blood components, and, thanks to the specific density of the separating gel, isolate the platelets and plasma in the upper part of the tube. Anticoagulation with sodium citrate has two advantages. It has no ancillary effects on the patient and is completely reversible.
It is therefore possible to induce coagulation and prepare RegenPRP in the form of platelet gel, platelet-enriched autologous fibrin sealant or fibrin clot. To activate the coagulation of RegenPRP, autologous thrombin-rich serum (called ATS) is prepared from the patient's blood using a RegenATS tube.®. This tube also separates blood components, but as it contains no anticoagulant, the plasma isolated above the separating gel coagulates.
Serum extracted from coagulum contains all the enzymes of the coagulation cascade in their active form, including trombin, the enzyme that converts soluble plasma fibrinogen into fibrin. polymerizes and forms the clot.
When RegenPRP is combined with ATS, these enzymes counterbalance the effect of sodium citrate and physiologically induce coagulation of RegenPRP. (Figure 2 and Figure 3).
Autologous thrombin-activated RegenPRP, in the form of platelet gel or autologous fibrin sealant, offers a promising approach to stimulating tissue healing and regeneration in a variety of clinical applications, from wound healing to promoting tissue rejuvenation.
The aim of the 2017 study [1] was to determine whether RegenPRP in the form of platelet-enriched autologous fibrin glue could accelerate and improve wound healing after skin reconstruction in cases of tissue loss secondary to soft tissue infection.
Figure 3. RegenBCT tubes® and RegenATS® to obtain a combination of RegenPRP and activated thrombin serum (ATS).
INDICATIONS ARE RECONSTRUCTION USING AUTOLOGOUS SKIN GRAFTS FOR LOSS OF SUBSTANCE SECONDARY TO NECROTIZING FASCIITIS OR FOURNIER'S GANGRENE
Twenty-seven patients were randomized into two groups:
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Experimental treatment (14 patients): Reconstruction with application of RegenPRP glue (Figure 4 and Figure 5);
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Conventional treatment (13 patients): Reconstruction without application of RegenPRP glue.
The average time to complete healing was significantly shortened by almost 50 % when autologous fibrin sealant was combined with the skin graft compared with the skin graft alone (Figure 6).
In the study group, patients achieved a skin graft success rate of 89.6 ± 13.2 %, whereas in the control group, graft success was 76.8 ± 11.5 % (p=0.01**) (Figure 7).
Figure 4. Patient 1, a 54-year-old man with extensive skin necrosis (2 % of total body surface area) located on the left thigh.
A thorough and precise surgical debridement of all necrotic tissue was performed in the operating room. A. After wound bed preparation, RegenPRP adhesive was sprayed onto the wound bed and then applied to the wound. B. on the skin graft after fixation with staples.
shows wound healing 2 days after skin grafting. D. Complete healing was achieved 21 days after skin grafting. The patient is discharged with the possibility of mobilization
5 days after skin reconstruction.
Figure 5. Patient 2, a 44-year-old man with Fournier's gangrene. Aggressive debridement of necrotic tissue was performed in the operating room.
After wound bed preparation, autologous fibrin sealant enriched with platelets was sprayed onto the wound bed and then applied to the wound. B. on the skin graft after fixation with staples.
The figure shows wound healing 2 weeks after skin grafting. D. Complete healing was achieved 45 days after skin grafting.
Figure 6. Average healing time for the control group and the experimental group.
Figure 7. Percentage of skin graft take at 7 days for the control group and the study group with autologous glue application (experimental group).
Figure 8. Preparation and application of RegenPRP glue between wound edges prior to suturing.
PLATELET-ENRICHED AUTOLOGOUS FIBRIN SEALANT IN WEIGHT LOSS SURGERY AND BREAST REDUCTION
The aim of this study [2] was to evaluate the adhesive action of platelet-enriched autologous glue obtained by combining RegenPRP with ATS (RegenPRP glue) in plastic surgery, and to analyze its impact on post-operative complications.
Modern plastic surgery is constantly striving to improve clinical outcomes while minimizing risks for patients. With this in mind, there is growing interest in the use of innovative products such as RegenPRP glue as a potential tool for optimizing healing and reducing post-operative complications (Figure 8).
- Fifty-one patients undergoing breast reduction surgery, abdominoplasty, thigh lift (vertical and horizontal) or brachioplasty with RegenPRP glue applied to the entire surface of the subcutaneous tissue at the time of suturing were included in this study.
- A retrospective control group operated on by the same surgeons using the same techniques but without application of RegenPRP glue was used as a comparator.
RegenPRP glue reduced the incidence of seroma formation after limb lifts, and prevented hematomas after breast reduction surgery (Table 2 and Table 3).
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Preparation and application of RegenPRP glue between wound edges prior to suturing. |
| Control Group | PRP Group | P* |
Seroma | 3/8 | 0/10 | 0.03* |
Hematoma | 1/8 | 0/10 | 0.44* (NS) |
Table 2. Postoperative complications of limb-lift patients at 1 month. P*, Fischer's exact test, P < 0.02; NS, not significant.
Control Group (n = 8) | PRP Group (n = 25) | P* | |
Seroma | 0/8 | 0/10 | 0.03* |
Hematoma | 2/8 | 0/10 | 0.44* (NS) |
Table 3. Postoperative complications in patients after breast reduction at 1 month. P*, Fischer's exact test, P < 0.05.
AUTOLOGOUS REGEN PRP AS AN ADJUVANT FOR THE TREATMENT AND PREVENTION OF KELOID SCARS
After conventional treatments, keloid scars show varying degrees of recurrence. The aim of this study [3] was to evaluate the efficacy and safety of RegenPRP in the treatment of postoperative keloid scars refractory to conventional treatments.
This prospective pilot study involved 17 patients with keloid scars who failed to respond to 4 cortisone injections or radiotherapy after extra-lesional keloid resection.
RegenPRP was injected intraoperatively and then 3 times at 1-month intervals.
- The primary endpoint was complete remission of keloid scars 2 years after treatment.
- The severity of scar pruritus was recorded before and after treatment. Nine keloid scars (53 %) were completely resolved at 2 years (Figure 9).
- Three patients (18 %) showed partial relapse, and 5 (29 %) relapsed completely after treatment.
- The pruritus severity score was significantly lower at 2 years than at baseline (1.33 ± 0.97 before treatment and 0.40 ± 0.63 at 2 years, P < 0.003) (Figure 10).
- The mean Vancouver Scar Scale score was significantly improved (8.18 ± 2.38 before treatment and 3.82 ± 1.98 at 2 years, P < 0.001) (Figure 11).
RegenPRP injection is a safe and effective adjunctive treatment to resection for keloid scars that are refractory to conventional therapy.
Figure 9. Two clinical cases of keloid scars in the earlobe that did not relapse. Appearance of keloid scar before treatment (A and C) and at 2 years (B and D).
Figure 10. Significant reduction in pruritus severity 2 years after treatment.
Figure 11. Significant decrease in VSS score showing improved scar quality 2 years after treatment.
REGEN PRP COMBINED WITH HYALURONIC ACID FOR FACIAL SKIN REJUVENATION
Cellular Matrix kits® are Class III medical devices enabling the preparation of RegenPRP combined with hyaluronic acid (HA) in a closed circuit.
The aim of this study [4] was to assess the clinical benefit of the RegenPRP-AH combination prepared with Cellular Matrix® and used in mesotherapy for skin rejuvenation (Figure 12).
This was a prospective study involving 31 patients.
- The primary endpoint was to measure the effectiveness of RegenPRP-AH in improving facial appearance, as assessed by the validated FACE-Q scale.
- The secondary endpoint was to measure the effectiveness of RegenPRP-AH in improving skin texture in terms of elasticity and firmness. Follow-up was carried out at 1, 3 and 6 months after 3 injection sessions (Figure 13 and Figure 14).
A significant improvement in FACE-Q scores after three treatment sessions and a significant improvement in skin elasticity parameters were observed.
All adverse events were reversible and well tolerated (Figure 15).
Figure 12. Preparation of RegenPRP associated with HA using Cellular Matrix tubes® BCT-HA.
Figure 13. Illustration of RegenPRP-AH injection techniques.
Figure 14. Photos of a patient before and after three RegenPRP-AH injection sessions.
Figure 15. Evaluation of the FACE-Q satisfaction scale for facial appearance.
REGENPRP-AH INJECTION FOR THE TREATMENT OF VULVOVAGINAL DRYNESS IN WOMEN
Around 50-70 % of breast cancer survivors are affected by one or more symptoms of vulvovaginal atrophy (VVA).
The RegenPRP-HA combination prepared with Cellular Matrix® technology (Figure XVI) may be an alternative therapy for the treatment of VTA in postmenopausal women with a history of breast cancer who cannot undergo hormone therapy.
Figure 16. Platelet-rich plasma with hyaluronic acid gel (PRP-HA) prepared with Cellular Matrix tube® BCT- HA. After centrifugation, the blood is fractionated: red blood cells are trapped under the separating gel, and platelets and plasma on top. Hyaluronic acid is placed on top of the plasma. By gently inverting the tube several times, the platelets are resuspended in the plasma, and the hyaluronic acid is added to the plasma. homogenization from acid hyaluronic with the RegenPRP. About 4 mL of RegenPRP-HA mixture is obtained for each tube.
- For this study [5], we recruited 20 postmenopausal breast cancer survivors with VTA and a score < 15 on the Gloria test.
- The Bachman Vaginal Health Index (VHI) comprised five elements, including: vaginal pH, elasticity, fluid volume (secretions), epithelial integrity and moisture.
We administered intramucosal injections of RegenPRP-HA (Figure 17) and performed clinical assessments at 0, 1, 3 and 6 months.
- The primary endpoint was the evaluation of vulvovaginal mucosal changes using the VHI. The secondary endpoint was the assessment of dyspareunia and sexual dysfunction on the basis of the Female Sexual Distress (FSD) score.
- All participants (20 women) showed improvement in clinical symptoms of vaginal dryness and dyspareunia.
- The VHI score showed a significant increase at 1, 3 and 6 months, rising from a baseline (pre-treatment) total score of 10.7 ± 2.12 to 20.75 ± 4.8 (P < 0.0001) at 6 months.
- An improvement in vaginal epithelial integrity and hydration has been reported.
A VHI score > 15 showed a positive therapeutic result. The pH showed a significant decrease (Figure 18).
Figure 17. A 56-year-old woman with episiotomy sequelae. Mucosal and submucosal injections of 4 mL RegenPRP-HA with a 27G needle into the vaginal wall (posterior) and vestibule (under scars and fissure).
Figure 18. A. Mean change in IHV and percentage improvement versus baseline (pre-treatment) up to 1 to 3 and 6 months post-treatment. B. Time trend in mean pH from baseline (pre-treatment) to 1 to 3 and 6 months post-treatment.
Figure 19. Female Sexual Distress (FSD) and percentage improvement from baseline (pre-treatment) at 1, 3 and 6 months post-treatment. Results represent mean standard deviation (SD). P < 0.04, P < 0.0001.
The FSD score decreased significantly over the course of the study, from an initial score of 36.35 ± 2.53 before treatment to 30.15 ± 2.47 6 months after treatment, representing an improvement of 17 % (P < 0.0001). No adverse events were reported (Figure 19).
EFFECT OF PRP ON THE POTENTIATION OF MESENCHYMAL STEM CELLS IN SKIN HEALING
Chronic and acute non-healing wounds represent a major public health problem, and replacing skin lesions with newly regenerated skin is a challenge.
- Mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) were tested separately in an attempt to regenerate lost skin.
- However, these treatments were often ineffective in achieving complete wound healing.
- Further studies suggest that PRP could be used in combination with MSCs to enhance the efficacy of cell therapy on tissue repair. However, systematic studies related to the effects of PRP on MSC properties and their ability to rebuild the skin barrier are lacking.
The model used in our study [6] was 6-week-old immunocompetent male mice (C57BL/6JRj) with 4 full-thickness wounds (Figure 20).
We evaluated the skin repair capacity of a treatment combining human-derived adipose MSCs and human RegenPRP compared with treatment with saline, RegenPRP alone or MSCs alone.
Figure 20. A. Experimental model of 6-week-old, immunocompetent male C57BL/6JRj mice.
Experimental diagram - zone 1: saline injection, zone 2: PRP 20% + CaCl2 10% injection, zone 3: MSC injection, zone 4: PRP 20% + CaCl2 10% + MSC injection.
Macroscopic analysis was performed to measure the percentage of skin regeneration for the different conditions and at different times (D3-J7).
- The percentage of skin regeneration for control (saline), PRP alone, MSC alone and MSC combined with PRP conditions was assessed at D3, D7 and D10.
- The results show that RegenPRP alone significantly increases skin regeneration, from 8 % for the control to 23 % for the PRP group (p < 0.001).
- At D3, MSC alone and combined with PRP very significantly increased skin regeneration compared with control p < 0.0001. The percentage of skin regeneration increased from 8 % for CLT to 32 % for MSC alone and 45 % for MSC+PRP.
- At D7, the RegenPRP-CSM combination significantly increased skin regeneration compared with the control (p<0.001).
- Skin regeneration increased from 43 % in the control group to 80 % in the MSC+PRP group.
- A significant increase was also observed in the MSC+PRP condition compared with the MSC alone condition (p < 0.01) and the MSC alone condition compared with the control (p < 0.05) (Figure 22A and B).
- At D10, no significant difference was observed between the different conditions.
Comparison of total healing times, corresponding to total epidermalization of the lesion (complete closure), shows that the MSC+PRP combination results in a significant reduction in total healing time compared with the control (P<0.01) (Figure 22C).
An analysis of biomechanical skin parameters was carried out after complete healing at D15 in order to compare skin elasticity under the different conditions.
The results of the gross elasticity (R2) measurement show a significant improvement in the PRP, CSM and CSM+PRP conditions compared with the control group. There is a greater improvement in the R2 parameter in the CSM+PRP condition compared with the other conditions (Figure 21).
Figure 21. Biomechanical parameters R2 (gross elasticity), R5 (net elasticity), R6 (viscoelasticity ratio) and R7 (biological elasticity) of the mouse healed wound after treatment with saline, PRP, hMADS cells or hMADS cells in combination with PRP, respectively. (a - d). Results represent mean ± SD obtained from n = 10 mice. ***p < 0,001, ****p < 0,0001.
Figure 22. The treatment PRP improves the potential from healing of CSM human (cells hMADS) following grafting into mouse wounds.
Closure rates of wounds treated with saline, PRP, hMADS cells or hMADS cells in combination with PRP at 3, 7 and 10 days following surgery. B. Representative photograph of the back of a mouse with skin wounds at 3 and 7 days after wounding and treatment with saline, PRP, hMADS cells or hMADS cells in combination with PRP, respectively. C. Wound healing times in mice treated with saline, PRP, hMADS cells or hMADS cells in combination with PRP, respectively. *p < 0,05, **p < 0,01, ***p < 0,001.
Figure 23. Hematoxylin-eosin staining of the dermis.
Histology shows that in the MSC condition, in association with PRP, the dermis is better organized, with more collagen fibers and myofibrils, and a greater number of capillaries, reflecting neoangiogenesis.
- Analysis of epidermal and dermal thickness showed no qualitative difference (Figure 23).
- Capillary density of biopsies at D3 and D7 after hMADS MSC transplantation was determined after labeling of microvessels with isolectin B4.
Isolectin B4 immunohistochemistry showed a highly significant difference in capillary density for MSC grafts alone and MSC+PRP (p<0.001***) at D3.
- A significant difference was observed between lesions treated with MSCs alone and those treated with PRP at D3 (p<0.05*).
- At D3, MSC+PRP significantly increased capillary density and thus angiogenesis compared with the MSC alone group (p < 0.05*) and lesions treated with PRP alone (p < 0.0001****).
- At D7, a significant increase in capillaries was only observed for MSC+PRP lesions compared with the control (saline treatment) (p < 0.05*) (Figure 24).
To demonstrate activation of the MSC secretome in the presence of PRP, we analyzed the transcriptional expression of VEGF, HGF and SDF-1 genes by quantitative RT PCR from biopsies of MSCs grafted naive or in the presence of 20% PRP, obtained after sacrifice of the mice on D1 and D3.
- The results summarized in the figure show significant activation of the MSC secretome in the presence of PRP at 20 % compared with naive cells, with a significant increase in the transcriptional expression of the 3 genes analyzed, VEGF, HGF and SDF-1 at D1 and D3.
- Thus the pro-angiogenic factors VEGF and HGF are significantly increased at D1 (p < 0.0001****, p < 0.001*** respectively) and D3 (p < 0.001***, p < 0.01** respectively) in biopsies treated with MSC+PRP compared with MSC alone.
- In parallel, the chemoattractant factor SDF-1 is significantly increased at D1 (p < 0.01*) and D3 (p < 0.01*) in biopsies treated with MSC+PRP compared with MSC alone.
- These results confirm those obtained in vivo showing that PRP increases the pro-angiogenic and chemoattractant capacity of MSCs (Figure 25).
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Figure 25. Relative transcriptional expression of human VEGF and human SDF-1 in mouse wounds corresponding to expression in hMADS cells grafted in combination with PRP compared with expression in hMADS cells grafted alone, at 1, 3 and 7 days post-wounding. Results represent the mean ± SD obtained from at least n = 5 mice per group. *p < 0,05, **p < 0,01, ***p < 0,001, ****p < 0,0001.
RESEARCH PROJECT: A PROSPECTIVE PILOT STUDY OF THE NASOLABIAL FOLD
Figure 26. RegenMatrix device®.
The Regen Matrix medical device® (under development, not yet certified) is an extension of the Cellular Matrix®.
The Regen Matrix® device combines the benefits of cross-linked hyaluronic acid (XLHA) with RegenPRP in a single, closed-circuit injection.
- When RegenPRP and XLHA are used in combination, the desired effects are enhanced and prolonged. XLHA creates an optimal "structural matrix" in which platelets gradually release their growth factors.
- The tube contains 2 mL of cross-linked HA at a concentration of 20 mg/ml (40 mg in total), produced by bacterial fermentation, without animal proteins (Figure XXVI).
REFERENCES
- Hersant, B., et al. Autologous Platelet-Rich Plasma/Thrombin Gel Combined with Split-Thickness Skin Graft to Manage Postinfectious Skin Defects: A Randomized Controlled Study. Adv Skin Wound Care 2017; 30(11): 502-8.
- Hersant, B., et al. Efficacy of Autologous Platelet-rich Plasma Glue in Weight Loss Sequelae Surgery and Breast Reduction: A Prospective Study. Plast Reconstr Surg Glob Open 2016; 4(11): e871.
- Hersant, B., et al. Efficacy of Autologous Platelet Concentrates as Adjuvant Therapy to Surgical Excision in the Treatment of Keloid Scars Refractory to Conventional Treatments: A Pilot Prospective Study. Ann Plast Surg 2018; 81(2): p. 170-5.
- Hersant, B., et al. Efficacy of autologous platelet-rich plasma combined with hyaluronic acid on skin facial rejuvenation: A prospective study. J Am Acad Dermatol 2017; 77(3): 584-6.
- Hersant, B., et al. Efficacy of injecting platelet concentrate combined with hyaluronic acid for the treatment of vulvovaginal atrophy in postmenopausal women with history of breast cancer: a phase 2 pilot study. Menopause 2018; 25(10): 1124- 30.
- Hersant, B., et al. Platelet-Rich Plasma Improves the Wound Healing Potential
