Leukocyte- And Platelet-rich Fibrin Research Articles

Can platelet-rich fibrin act as a natural carrier for antibiotics delivery? A proof-of-concept study for oral surgical procedures

ObjectivesEvaluate the role of platelet-rich fibrin (PRF) as a natural carrier for antibiotics delivery through the analysis of drug release and antimicrobial activity.Materials and methodsPRF was prepared according to the L-PRF (leukocyte- and platelet-rich fibrin) protocol. One tube was used as control (without drug), while an increasing amount of gentamicin (0.25 mg, G1; 0.5 mg, G2; 0.75 mg, G3; 1 mg, G4), linezolid (0.5 mg, L1; 1 mg, L2; 1.5 mg, L3; 2 mg, L4), vancomycin (1.25 mg, V1; 2.5 mg, V2; 3.75 mg, V3; 5 mg, V4) was added to the other tubes. At different times the supernatant was collected and analyzed. Strains of E. coli, P. aeruginosa, S. mitis, H. influenzae, S. pneumoniae, S. aureus were used to assess the antimicrobial effect of PRF membranes prepared with the same antibiotics and compared to control PRF.ResultsVancomycin interfered with PRF formation. Gentamicin and linezolid did not change the physical properties of PRF and were released from membranes in the time intervals examined. The inhibition area analysis showed that control PRF had slight antibacterial activity against all tested microorganisms. Gentamicin-PRF had a massive antibacterial activity against all tested microorganisms. Results were similar for linezolid-PRF, except for its antibacterial activity against E. coli and P. aeruginosa that was comparable to control PRF.ConclusionsPRF loaded with antibiotics allowed the release of antimicrobial drugs in an effective concentration. Using PRF loaded with antibiotics after oral surgery may reduce the risk of post-operative infection, replace or enhance systemic antibiotic therapy while preserving the healing properties of PRF. Further studies are needed to prove that PRF loaded with antibiotics represents a topical antibiotic delivery tool for oral surgical procedures.

Histological, Immunohistochemical, Biomechanical, and Wettability Evaluations of the Leukocyte- and Platelet-Rich Fibrin Membranes Derived from Canine Blood.

This study aimed to perform histological, immunohistochemical, biomechanical, and wettability assessments of leukocyte- and platelet-rich fibrin (L-PRF) membranes obtained from the blood of healthy dogs. Ten client-owned Labrador Retriever dogs were enrolled. Blood samples were obtained from the external jugular vein using a vacuum tube without anticoagulant, which was immediately centrifuged at 400g for 12 min in a dedicated centrifuge. The L-PRF clot was removed from the tube, and the red clot was released from the buffy coat using a spatula. The membrane was produced using a PRF box. Histological examination identified the three portions of the L-PRF membranes. The first portion was composed mainly of red blood cells with the presence of a low number of leukocytes among them. The second portion was composed of white blood cells, mainly neutrophils. The third portion was composed of the fibrin network which was characterized by acidophilic staining. The immunohistochemical analysis showed that vascular endothelial growth factor and platelet-derived growth factor were expressed in all samples at different intensities, both in cellular components and fibrin mesh. The tensile test and wettability assessments were measured in membranes 30 min and 3 h after production. The 30 min L-PRF membranes supported twice the ultimate tensile strength compared to 3 h L-PRF membranes. The wettability of the 30 min sample membranes was statistically higher than the 3 h sample membranes. In conclusion, the centrifugation protocol allowed production of the L-PRF membrane using canine blood and this was confirmed by histological and immunohistochemical analysis. The mechanical resistance and wettability of the L-PRF membrane were significantly reduced over time.

Comparative Evaluation of Titanium-platelet-rich Fibrin and Leukocyte-platelet-rich Fibrin – A Scanning Electron Microscopic Study

ABSTRACT Background: Titanium-prepared platelet-rich fibrin (T-PRF), a new autologous platelet-rich fibrin (PRF) product, is based on the hypothesis that titanium tubes may be more effective in activating platelets when compared with Choukroun’s leukocyte and PRF (L-PRF) method. In this study, we aimed to define the structural characteristics under scanning electron microscopy of T-PRF and compare it with L-PRF. Materials and Methods: Blood samples were collected from 10 healthy volunteers. The blood samples were drawn using a 10 ml syringe. A volume of 9 ml was transferred to a dry glass tube, and 9 ml was transferred to a titanium tube and centrifuged. Half of each clot (i.e. after centrifugation, the T-PRF and L-PRF clots were collected) samples were processed with a scanning electron microscope analysis. Results: The T-PRF samples seemed to have a highly organized network with continuous integrity compared to the other L-PRF samples. Conclusion: Histomorphometry analysis showed that the T-PRF fibrin network covers a larger area than the L-PRF fibrin network, and also, fibrin seemed thicker in the T-PRF samples. The platelet activation by titanium seems to offer some high characteristics to T-PRF. CLINICAL RELEVANCE TO INTERDISCIPLINARY DENTISTRY Titanium-prepared platelet-rich fibrin (T-PRF), A new autologous platelet fibrin rich product can be used in regeneration of pulpal and periodontal tissues.

The Role of Leukocyte and Platelet-Rich Fibrin in Enhancing the Healing of Extraction Sockets: An Overview of the Literature

Introduction: Leukocyte and platelet-rich fibrin (L-PRF) is an emerging material in dentistry, however, there are controversies surrounding its effectiveness. Despite the amount of literature available, debates regarding its effect continue. This review aims to summarize and clarify the data surrounding the use of L-PRF in promoting the healing of extraction sockets, which may offer a better outcome for future treatments. Purpose: The purpose of this review is to evaluate the current literature on the use of L-PRF in promoting the healing of extraction sockets, and to provide a comprehensive overview of the available evidence. Methods: A comprehensive computer-based search of databases such as PubMed, Medline, and Cochrane Library was conducted. Results: The results of this review suggest that L-PRF has shown promise in promoting early healing of extraction sockets, but the evidence for its effectiveness over a longer period is limited. Conclusion: Although L-PRF has shown promising results in the early healing periods, its effectiveness over a longer healing period cannot be confirmed based on the available data. More clinical trials with standardized protocols and consistent measurement methods are needed to establish the role of L-PRF in enhancing the healing of extraction sockets.

Effect of Leucocyte and Platelet Rich Fibrin (L-PRF) in Postoperative Sequelae of Impacted Mandibular Third Molar Surgery - A Split Mouth Randomized Controlled Trial

Effect of Leucocyte and Platelet Rich Fibrin (L-PRF) in Postoperative Sequelae of Impacted Mandibular Third Molar Surgery - A Split Mouth Randomized Controlled Trial

Innovative methods and developments in oral care. L-PRF: from evidence to clinical use

Leukocyte- and platelet-rich fibrin (L-PRF) is a fully autologous biomaterial made from venous blood drawn from the patient. Due to its regenerative potency, antibacterial capacity, and analgesic activity, L-PRF can be used during surgical procedures as a sole biomaterial or as a bioactive additive along with other natural and synthetic biomaterials. There is sufficient scientific evidence available for applying L-PRF for various indications. The preparation protocol of L-PRF is simple, inexpensive and not time-consuming, making it possible to implement it in the daily practice. The type of centrifuge and blood collection tubes used, the settings of the centrifuge (rotation time and force) as well as the time between the different steps in the protocol have an influence on the biological and mechanical properties of L-PRF.

The impact of gender and peripheral blood parameters on the characteristics of L-PRF membranes.

The purpose of this study was to evaluate the impact of gender and peripheral blood parameters on the characteristics of Leucocyte-and Platelet-Rich Fibrin (L-PRF) membranes and to describe histologically three different zones of L-PRF membranes. Blood was collected from twenty healthy donors (10 men and 10 women). Peripheral blood parameters including leucocyte and platelet counts, and fibrinogen levels were recorded. L-PRF membranes were prepared to quantify the release of growth factors (PDGF, VEGF, BMP-2, and BMP-9) at 1, 2, 3 and 7 days and for histological examination. Three zones within each L-PRF membrane (face, body, and tail) were analysed separately, quantifying the area of leucocytes, platelets, and fibrin in percentage. The Young's modulus of the membranes was also considered (during tensile and compression tests). Women had significantly higher fibrinogen levels in their peripheral blood, and a higher release of BMP-9, whereas men showed a significantly higher Young's modulus in compression tests. The histology revealed significant differences in cellular content and fibrin concentration between the 3 areas, with the face being biologically the richest. Several factors influenced the final characteristics of L-PRF membranes. These need to be taken into consideration when interpreting the results of research, but especially in clinical practice.

Comparison of physical, mechanical and biological effects of leucocyte-PRF and advanced-PRF on polyacrylamide nanofiber wound dressings: In vitro and in vivo evaluations.

Platelet-rich fibrin (PRF) is extracted from the blood without biochemical interference and, also, with the ability of a long-term release of growth factors that can stimulate tissue repair and regerenation. Here, leucocyte- and platelet-rich fibrin (L-PRF) and advanced platelet-rich fibrin (A-PRF) were extracted and utilized for the creation of nanofibers containing polyacrylamide (PAAm), PAAm / L-PRF and PAAm / A-PRP through electrospinning processing technique. The effect of the type of PRF on the physical, mechanical and biological properties of the resultant nanofiberous wound dressings are thoroughly evaluated. The results presented in the current study reveals that the fiber diameter is grealtly reduced through the utilization of L-PRF. In addition, mechanical property is also positively affected by L-PRF and the degradation rate is found to be higher compared to A-PRF group. The L929 cells proliferation and adhesion, angiogenesis potential and wound healing ability was significantly higher in PAAm/A-PRF nanofibers compared to pure PAAm and PAAm/L-PRF nanofibers owed to the release of vascular endothelial growth factor (VEGF) and platelet derived growth factor (PDGF). Overall, the utilization of L-PRF or A-PRF can improve the physical, mechanical and biological behavior of nanofiber making them an ideal candidate for wound dressings, with the emphasis on the skin tissue repair and regeneration applications.

Suburethral implantation of autologous regenerative cells for female stress urinary incontinence management: Results of a pilot study

ObjectivesTo assess the feasibility and the safety of treating female stress urinary incontinence (SUI) with suburethral implantation of a mixture of the stromal vascular fraction from adipose tissue and leukocyte-and platelet-rich-fibrin. MethodsPatients with SUI were treated with a mixture of stromal vascular fraction and leukocyte-and platelet-rich fibrin. The stromal vascular fraction was obtained from enzymatic digestion of autologous adipose-tissue and added to an leukocyte-and platelet-rich-fibrin membrane. The mixture was transvaginally implanted into the suburethral area. A fraction of the Stromal vascular fraction sample was used for cellular characterization. Patients were followed for 9 months. Every 3 months, the patients were clinically evaluated with a cough- stress test and a validated-questionnaire. An MRI was performed preoperatively and 3 months after the procedure to assess tissue changes. ResultsTen patients received the surgical procedure. The validated-questionnaire revealed a subjective SUI improvement in nine patients 3 months after the operation and in seven patients 9 months after the operation. Eight, six, and four patients achieved a negative cough-stress test 3, 6 and 9 months post-injection, respectively. Flow cytometric analysis of stromal vascular fraction cell phenotypes revealed predominantly mesenchymal and endothelial cell heterogeneity. In total, we injected 0,18 × 106 to 13,6 × 106 cells. No adverse events were observed peri- or postoperatively. ConclusionThese preliminary results suggest that the suburethral implantation of a combination of SVF and l-PRF is a feasible and safe modality for treating female SUI. However, evidence is lacking and further research are needed to clarify the respective roles of SVF and l-PRF in female SUI treatment.

Effect of leukocyte and platelet-rich fibrin on free gingival graft healing: A clinical and histological study in rabbits.

Background. Recently, the use of leukocyte- and platelet-rich fibrin (L-PRF) has been recommended due to the presence of various growth factors to increase the success of free gingival grafts (FGG). This study evaluated the effect of using L-PRF in the healing of FGG in rabbits. Methods. Twenty rabbits were randomly divided into two groups. In each group, FGG was performed in two separate sites with or without L-PRF. One of these groups was sacrificed on the 7th day and the other on the 28th day and analyzed in terms of clinical indices, including wound healing, gingi-val thickness (GT), and keratinized tissue width (KTW). Then histologic sections were obtained and stained for type and degree of inflammation and rate of vascular formation analysis. SPSS 22 was used for statistical analysis. Results. The extent of changes in GT, KTW, wound healing index, and vascular formation between the test and control groups was not statistically significant. The difference in the type of inflammation was significant only between the -7day and -28day control groups (P=0.003). The degree of inflammation between the -7day test group and the -28day control group, as well as the -7day and -28day control groups, were statistically significant (P=0.011 and P=0.002, respectively). Conclusion. Using L-PRF with FGG could improve FGG healing compared to using FGG alone, but the results were not statistically significant.

Application of Fibrin Associated with Photobiomodulation as a Promising Strategy to Improve Regeneration in Tissue Engineering: A Systematic Review.

Fibrin, derived from proteins involved in blood clotting (fibrinogen and thrombin), is a biopolymer with different applications in the health area since it has hemostasis, biocompatible and three-dimensional physical structure properties, and can be used as scaffolds in tissue regeneration or drug delivery system for cells and/or growth factors. Fibrin alone or together with other biomaterials, has been indicated for use as a biological support to promote the regeneration of stem cells, bone, peripheral nerves, and other injured tissues. In its diversity of forms of application and constitution, there are platelet-rich fibrin (PRF), Leukocyte- and platelet-rich fibrin (L-PRF), fibrin glue or fibrin sealant, and hydrogels. In order to increase fibrin properties, adjuvant therapies can be combined to favor tissue repair, such as photobiomodulation (PBM), by low-level laser therapy (LLLT) or LEDs (Light Emitting Diode). Therefore, this systematic review aimed to evaluate the relationship between PBM and the use of fibrin compounds, referring to the results of previous studies published in PubMed/MEDLINE, Scopus and Web of Science databases. The descriptors “fibrin AND low-level laser therapy” and “fibrin AND photobiomodulation” were used, without restriction on publication time. The bibliographic search found 44 articles in PubMed/MEDLINE, of which 26 were excluded due to duplicity or being outside the eligibility criteria. We also found 40 articles in Web of Science and selected 1 article, 152 articles in Scopus and no article selected, totaling 19 articles for qualitative analysis. The fibrin type most used in combination with PBM was fibrin sealant, mainly heterologous, followed by PRF or L-PRF. In PBM, the gallium-aluminum-arsenide (GaAlAs) laser prevailed, with a wavelength of 830 nm, followed by 810 nm. Among the preclinical studies, the most researched association of fibrin and PBM was the use of fibrin sealants in bone or nerve injuries; in clinical studies, the association of PBM with medication-related treatments osteonecrosis of the jaw (MRONJ). Therefore, there is scientific evidence of the contribution of PBM on fibrin composites, constituting a supporting therapy that acts by stimulating cell activity, angiogenesis, osteoblast activation, axonal growth, anti-inflammatory and anti-edema action, increased collagen synthesis and its maturation, as well as biomolecules.

A Comprehensive Review on Efficacy of Leukocyte and Platelet-rich Fibrin (L-PRF) Administration in Reconstruction of Oral and Maxillofacial Structures

Several studies have shown that growth factors are effective in soothing inflammatory processes involved in the initial hemostasis phase of wound healing. Leukocyte and platelet-rich fibrin (L-PRF) is the second generation of concentrated platelets that have been shown to stimulate the proliferation of osteoblasts, gingival fibroblasts, and periodontal ligament cells. Rich platelet components have an immune function and antibacterial properties, which play a critical role in the wound healing process. The present study aimed to review the reported documents on L-PRF administration in regeneration and wound healing of oral and maxillofacial surgeries. The research was performed using well-known medical databases to find controlled clinical trials that used L-PRF as a cotreatment strategy during oral and maxillofacial surgeries. Eligible articles published between 2000 and 2022 in PubMed, Web of Science, Science Direct, and Scopus databases were selected for this review. According to the results of this study, L-PRF has significant effectiveness as a maxillofacial regenerative therapy by improving para-clinical parameters after surgery. However, more clinical practice and additional research are essential for the development of this complementary treatment method

Increased local concentrations of growth factors from leucocyte‐ and platelet‐rich fibrin do not translate into improved alveolar ridge preservation: An intra‐individual mechanistic randomized controlled trial

Leucocyte- and platelet-rich fibrin (L-PRF) has been tested for enhancing alveolar ridge preservation (ARP), but little is known about the local release profile of growth factors (GF), and the clinical equipoise related to its efficacy remains. This study compared the patterns of GF release, early soft tissue healing, and alveolar ridge resorption following unassisted healing and L-PRF application in non-molar extraction sockets. Atraumatic tooth extraction of two hopeless teeth per patient was followed by unassisted healing or L-PRF placement to fill the socket in 18 systemically healthy, non-smoking subjects. This intra-individual trial was powered to assess changes in horizontal alveolar ridge dimensions 1 mm below the crest of alveolar bone. GF concentrations in wound fluid were assessed with a multiplex assay at 6, 24, 72, and 168 h. Early healing was evaluated with the wound healing index and changes in soft tissue volumes on serial digital scans. Hard tissue changes were measured on superimposed CBCT images after 5months of healing. L-PRF resulted in higher GF concentrations in wound fluid (WF) than in the control, but no differences in release patterns or time of peak were observed. No inter-group differences in early healing parameters were observed. Alveolar bone resorption was observed in both groups. No significant inter-group differences were observed in hard tissue healing 1, 3, or 5 mm apical to the original bone crest or in the ability to digitally plan a prosthetically guided implant with or without bone augmentation. L-PRF increased the GF concentrations in WF of extraction sockets without shifting the pattern observed in unassisted healing, while the increased delivery did not translate into clinical benefits in early wound healing or ARP. The current findings question the assumption that increased local concentrations of GF by L-PRF translate into improved clinical outcomes. Additional definitive studies are needed to establish the benefits of L-PRF in ARP (ClinicalTrials.gov NCT03985033).

In Vitro and Ex Vivo Kinetic Release Profile of Growth Factors and Cytokines from Leucocyte- and Platelet-Rich Fibrin (L-PRF) Preparations.

L-PRF is an autologous blood-derived biomaterial (ABDB) capable of releasing biologically active agents to promote healing. Little is known about its release profile of growth factors (GFs), cytokines, and MMPs. This study reported the in vitro and ex vivo release kinetics of GFs, cytokines, and MMPs from L-PRF at 6, 24, 72, and 168 h. The in vitro release rates of PDGF, TGF-β1, EGF, FGF-2, VEGF, and MMPs decreased over time with different rates, while those of IL-1β, IL-6, TNF-α, IL-8, and IL-10 were low at 6 h and then increased rapidly for up to 24 h and subsequently decreased. Of note, the release rates of the GFs followed first-order kinetics both in vitro and ex vivo. Higher rates of release were found ex vivo, suggesting that significant amounts of GFs were produced by the local cells within the wound. In addition, the half-life times of GFs locally produced in the wound, including PDGF-AA, PDGF-AB/BB, and VEGF, were significantly extended (p < 0.05). This work demonstrates that L-PRF can sustain the release of GFs and cytokines for up to 7 days, and it shows that the former can activate cells to produce additional mediators and amplify the communication network for optimizing the wound environment, thereby enhancing healing.

Complementarity of surgical therapy, photobiomodulation, A-PRF and L-PRF for management of medication-related osteonecrosis of the jaw (MRONJ): an animal study

BackgroundThis study aimed to evaluate the complementarity of surgical therapy, photobiomodulation (PBM), advanced platelet-rich fibrin (A-PRF), and Leukocyte and platelet-rich fibrin (L-PRF) for the management of medication-related osteonecrosis of the jaw (MRONJ).MethodsSixty rats underwent injection of zoledronate followed by left mandibular first and second molar extractions to induce MRONJ lesions. All rats were examined for the signs of MRONJ 8 weeks post-dental extraction. Forty-nine rats with positive signs of MRONJ were appointed to seven different groups as follows: control (Ctrl); surgery alone (Surg); surgery and PBM (Surg + PBM); surgery and A-PRF insertion (Surg + APRF); surgery and L-PRF insertion (Surg + LPRF); surgery, A-PRF insertion, and PBM (Surg + APRF + PBM); surgery, L-PRF insertion, and PBM (Surg + LPRF + PBM). Euthanasia was carried out 30 days after the last treatment session. The lesions' healing was evaluated clinically, histologically, and radiographically. Data were analyzed using STATA software version 14, and the statistical significance level was set at 5% for all cases.ResultsAccording to the present study, A-PRF and L-PRF treatment resulted in significant improvements in clinical, histological, and radiographical parameters compared to the Ctrl group (P < 0.05). The PBM also decreased wound dimensions and the number of empty lacunae compared to the Ctrl group (P < 0.05). Surg + APRF + PBM and Surg + LPRF + PBM were the only groups that presented a significantly higher mean number of osteocytes (P < 0.05). No significant differences were observed between A-PRF and L-PRF treatment groups (P > 0.05).ConclusionsSurgical resection followed by applying A-PRF or L-PRF reinforced by PBM showed optimal wound healing and bone regeneration in MRONJ lesions.

Production Protocol Standardisation, Macroscopic and Histological Evaluation, and Growth Factor Quantification of Canine Leukocyte-and Platelet-Rich Fibrin Membranes.

Leukocyte-Platelet-Rich Fibrin (L-PRF) is a second generation of platelet concentrates; it was widely used, as an autologous platelet-based wound sealant and hemostatic agent in surgical wound healing. L-PRF clot or membrane is a solid fibrin-based biomaterial, with a specific 3D distribution of the leukocytes and platelet aggregates. This biological scaffold releases growth factors (i.e., TGF- β1, PDGF-AB, VEGF) and matrix proteins (fibronectin, vitronectin and thrombospondin-1) during the healing process after the application. To the Authors' knowledge both in human and veterinary medicine a single standardised protocol was not reported. This prospective study aimed to apply Crisci's L-PRF protocol (which is characterised by 30” of acceleration, 2' at 2,700 rpm, 4' at 2,400 rpm, 3' at 3,000 rpm, and 36” of deceleration and arrest) sin canine species, evaluate macroscopically and histologically the L-PRF membranes obtained by using Wound Box to standardise the L-PRF protocol in dogs and to evaluate the clinical feasibility of using L-PRF membranes by quantitative in vitro analysis of growth factors over 7 days. One hundred twenty-eight dogs in good general condition with no history of recent NSAIDs intake (15 days of washout) and/or any medication or disease related to coagulation process met inclusion criteria and therefore were enrolled. We obtained 172 membrane L-PRF membranes by 86 dogs: half of them underwent macroscopic and histological analysis, the other 86 underwent ELISA analysis. The Wound Box gave a membrane of mean (±SD) length (cm), width (cm) and weight (g) of 1.97 (±0.89), 0.95 (±0.36), 0.46 (±0.20) respectively. Histology analysis confirmed a well-defined histoarchitecture with five layers reproducing density and distribution of blood cells in this biomaterial. Finally, the ELISA assay performed with 22 L-PRF membranes showed a peak in growth factors at 6 h after membrane production, followed by a decrease in release at 24 and 72 h and a second peak in release at 168 h after production. Statistical analysis of demographic variables (age, sex, and body condition score BCS) and the average of growth factors determined by the ELISA assay did not reveal statistical significance, except for the BCS factor compared with the production of VEGF. Our data confirm the effectiveness of this protocol and of Wound Box to produce L-PRF membranes in dogs.

Mechanical properties and cellular content of leukocyte- and platelet-rich fibrin membranes of patients on antithrombotic drugs.

The aim of this study was to examine the potential influence of antithrombotics on leukocyte- and platelet-rich fibrin (L-PRF) membranes. Tensile tests and cell counts were performed with L-PRF membranes originating from patients on anticoagulants and antiplatelets versus patients not taking antithrombotics. For the tensile tests, 13 control patients, 12 on anticoagulants, and 10 on antiplatelets donated blood. Compared to controls, membranes from anticoagulated donors were weaker (strength 0.57±0.24MPa vs. 0.80±0.27MPa, p=.03) and could not be stretched as far (1.8±0.3 vs. 2.1±0.3 times the initial length, p=.01). For the cell counting, 23 control patients, 16 on anticoagulants, and 16 on antiplatelets donated blood. The percentage of platelets was ±50% in the three groups. The percentage of leukocytes was lower in the anticoagulant group compared with controls (69±10% vs. 78±8%, p=.04). However, because of the unknown error of method, it is questionable whether the statistical significance is meaningful. There was no difference between membranes from the control group and the group on antiplatelets. Our results indicate that L-PRF membranes originating from patients on anticoagulants are weaker, stretch less far, and contain less leukocytes than L-PRF membranes of patients not taking these drugs.

Comparative ex-vivo evaluation of the expression of Vascular Endothelial Growth Factor (VEGF) in Leukocyte and Platelet-Rich Fibrin (L-PRF), injectable Platelet-Rich Fibrin (i-PRF), &amp; Stored Whole blood: An ELISA based observational study

Introduction: Platelets have a vital role in haemostasis, healing and secretion of growth factors. The biologically active peptides in platelets significantly modulate proliferation of human pluripotent stem cells including periodontal ligament cells, thus, aiding regeneration. Vascular Endothelial Growth Factor (VEGF) is an angiogenic growth factor that promotes healing. Objective: Evaluation of expression of VEGF levels in Leukocyte and Platelet-Rich Fibrin (L-PRF), injectable Platelet-Rich Fibrin (i-PRF, and stored whole blood.Methodology: Autologous blood was collected in sterile/citrated vacutainers by drawing peripheral blood samples from 10 healthy subjects. Each sample was divided into 3 groups (Stored Whole Blood, L-PRP & i-PRF). The samples were processed for preparation of Platelet Concentrates viz Leukocyte and Platelet-Rich Fibrin (L-PRF), injectable Platelet-Rich Fibrin (i-PRF) as per Choukroun’s protocols. Quantitative assessment of expression of VEGF was carried out with ELISA. Comparative statistical analysis was performed using SYSTAT-13 software.Results: Quantitative analysis of VEGF revealed higher levels in Leukocyte and Platelet-Rich Fibrin (L-PRF), injectable Platelet-Rich Fibrin (i-PRF) compared to those of stored whole blood groups.Conclusion: Leukocyte and Platelet-Rich Fibrin (L-PRF), injectable Platelet-Rich Fibrin (i-PRF) having increased expression of VEGF can be preferentially applied in regenerative periodontal treatment protocols.

The G-Force Conundrum in Platelet-Rich Fibrin Generation

Aim:A force of 400 g at 2700 revolutions per minute (RPM) results in an optimum leukocyte and platelet-rich fibrin (L-PRF). Most of centrifuges with varying characteristics generate a g-force in excess of 700 g at 2700 RPM. In this context, the study explores the effect of the original centrifugation protocol and a modified protocol tailor-made to lower the RPM to generate a g-force of ~ 400 g on platelet concentration, clot size and growth factors release in L-PRF prepared in two different commercially available centrifuges.Materials and Methods:Twenty five subjects each were assigned to the following groups; R1 and R2 where L-PRF was obtained from two laboratory swing-out centrifuges (Remi 8C® and Remi C854®, Mumbai, India), respectively. PRF was obtained from each subject within a group using two protocols; Original (O) protocol: conforming to the original centrifugation cycle (2700 RPM for 12 min) and Modified (M) protocol. Clot size, growth factor estimation, and platelet counts were measured at 20, 40, and 60 min from all the L-PRF clots, respectively.Results:At the third time period (40–60 min), there were no significant differences in clot sizes with the original protocol (P = 0.09), but a highly significant difference was noticed with the modified protocol in both the centrifuges (P = 0.001). Our results showed an increased concentration of vascular endothelial growth factor and epidermal growth factor with modified protocol than with original protocol with both the centrifuges (P = 0.001). By the end of second and third time periods, more platelet concentration was observed with modified protocol than with the original protocol in both the centrifuges (P = 0.001).Conclusion:This study infers that the centrifuge type and relative centrifugal force can affect the quality and quantity of cells and growth factors and an optimum relationship between g-force and RPM should be maintained to obtain L-PRF with adequate cell viability and optimum growth factor release.

The Adjunctive Use of Leucocyte- and Platelet-Rich Fibrin in Periodontal Endosseous and Furcation Defects: A Systematic Review and Meta-Analysis.

The aim of this systematic review of randomized controlled trials was to evaluate the adjunctive use of leucocyte- and platelet-rich fibrin (L-PRF) in periodontal endosseous and furcation defects, as compared without L-PRF. The endosseous defect group was subclassified into: L-PRF/open flap debridement (L-PRF/OFD) versus OFD, L-PRF/osseous graft (L-PRF/OG) versus OG, L-PRF/Emdogain (L-PRF/EMD) versus EMD, and L-PRF/guided tissue regeneration (L-PRF/GTR) versus GTR. The furcation defect group was subclassified into L-PRF/OFD versus OFD, and L-PRF/OG versus OG. Mean difference, 95% confidence intervals and forest plots were calculated for probing pocket depth (PPD), clinical attachment level (CAL) and radiographic defect depth (DD). Nineteen studies concerning systemically healthy non-smokers were included. The results of this systematic review and meta-analysis showed in two- and/or three-wall endosseous defects that the adjunctive use of L-PRF to OFD or OG was significantly beneficial for PPD reduction, CAL gain and DD reduction, as compared without L-PRF. Furthermore, the data showed that for two- and/or three-wall endosseous defects, the adjunctive use of L-PRF to GTR was significantly beneficial for CAL and DD improvement, whereas adding L-PRF to EMD had no significant effect, and that for class II furcation defects, the addition of L-PRF to OFD was significantly beneficial for PPD, CAL and DD improvement, whereas the addition of L-PRF to OG was significantly clinically beneficial. In conclusion, this systematic review and meta-analysis found that there was significant clinical and radiographic additive effectiveness of L-PRF to OFD and to OG in two- and/or three-wall periodontal endosseous defects of systemically healthy non-smokers, as compared without L-PRF.