Fibrin Membrane Research Articles

Cellular composition modifies the biological properties and stability of platelet rich plasma membranes for tissue engineering.

Scaffolds should provide structural support for tissue regeneration, allowing their gradual biodegradation and interacting with cells and bioactive molecules to promote remodeling. Thus, the scaffold's intrinsic properties affect cellular processes involved in tissue regeneration, including migration, proliferation, differentiation, and protein synthesis. In this sense, due to its biological effect and clinical potential, Platelet Rich Plasma (PRP) fibrin could be considered a successful scaffold. Given the high variability in commercial PRPs formulations, this research focused on assessing the influence of cellular composition on fibrin membrane stability and remodeling cell activity. The stability and biological effect were evaluated at different time points via D-dimer, type I collagen and elastase quantification in culture media conditioned by Plasma Rich in Growth Factors - Fraction 1 (PRGF-F1), Plasma Rich in Growth Factors - Whole Plasma (PRGF-WP) and Leukocyte-rich Platelet Rich Plasma (L-PRP) membranes, and by gingival fibroblast cells seeded on them, respectively. Ultrastructure of PRP membranes was also evaluated. Histological analyses were performed after 5 and 18 days. Additionally, the effect of fibrin membranes on cell proliferation was determined. According to the results, L-PRP fibrin membranes degradation was complete at the end of the study, while PRGF membranes remained practically unchanged. Considering fibroblast behavior, PRGF membranes, in contrast to L-PRP ones, promoted extracellular matrix biosynthesis at the same time as fibrinolysis and enhanced cell proliferation. In conclusion, leukocytes in PRP fibrin membranes drastically reduce scaffold stability and induce behavioral changes in fibroblasts by reducing their proliferation rate and remodeling ability.

Bacterial exclusion and wound healing potential of horizontal platelet-rich fibrin (H-PRF) membranes when compared to 2 commercially available collagen membranes.

The aim of the present study was to compare the barrier function during bacterial invasion and wound healing properties of 3 commonly used membranes including horizontal platelet-rich fibrin (H-PRF) against two commercially available resorbable collagen membranes. H-PRF membranes were prepared by collecting venous blood from 3 healthy volunteers using a 700g for 8-min centrifugation protocol followed by compression into membranes. To evaluate their barrier function, 3 groups (H-PRF membrane, collagen membrane A (Bio-Gide, Geistlich), collagen membrane B (Megreen, Shanxi Ruisheng Biotechnology Co) were placed between an inner chamber and outer chamber and inoculated with S. aureus. At 2h, 24h, and 48h post-inoculation, cultures from the inner and outer chambers were assessed for bacterial CFUs. Then, scanning electron microscope (SEM) was utilized to visualized the morphological destruction by bacteria of the inner and outer surfaces of the membranes. To assess the wound healing properties of each membrane, leachates from each group were applied to human gingival fibroblasts (HGF) and a scratch assay was performed at 24h and 48h. S. aureus showed a minimal bacterial attachment or invasion rate through either collagen membranes at 2h post-inoculation, yet over time demonstrated rapid degradation, especially on the rougher surface. While PRF demonstrated higher number of CFUs after 2h, no significant penetration/degradation of the H-PRF membranes was observed at 24h and 48h in the H-PRF group. Both collagen membranes demonstrated significant morphological changes 48h post-bacterial innoculation, while minimal obvious morphological changes were observed in the H-PRF group. The wound healing assay also demonstrated significantly better wound closure rates in the H-PRF group. H-PRF membranes exhibited better barrier function towards S. aureus over 2days of innoculation and better wound healing ability when compared to two commercially available collagen membranes. This study provides further evidence for the application of H-PRF membranes during guided bone regeneration by minimizing bacterial invasion. Furthermore, H-PRF membranes have significantly better ability to promote wound healing.

Development of a new plasma rich in growth factors membrane with improved optical properties.

The aim of the present work was to develop a fibrin membrane using plasma rich in growth factors (PRGF) technology with improved optical properties to be used for the treatment of ocular surface diseases. Blood was drawn from three healthy donors, and the volume of PRGF obtained from each donor was divided into two main groups: i) PRGF or ii) platelet-poor plasma (PPP). Each membrane was then used pure or diluted to 90 %, 80 %, 70 %, 60 % and 50 %. The transparency of each of the different membranes was evaluated. The degradation and morphological characterization of each membrane was also performed. Finally, a stability study of the different fibrin membranes was performed. The transmittance test showed that the fibrin membrane with the best optical characteristics was obtained after removal of platelets and dilution of fibrin to 50 % (50 % PPP). No significant differences (p>0.05) were observed between the different membranes in the fibrin degradation test. The stability test showed that the membrane at 50 % PPP retains its optical and physical characteristics after storage at -20 °C for 1 month compared to storage at 4°C. The present study describes the development and characterization of a new fibrin membrane with improved optical characteristics while maintaining mechanical and biological characteristics. The physical and mechanical properties of the newly developed membrane are preserved after storage for at least 1 month at -20 °C.

Use of platelet-rich fibrin (PRF) in the periodontology: A Review

Platelets play an essential role in wound healing and periodontal regeneration, so using platelet concentrates can speed up wound healing after periodontal treatment. Platelet-rich fibrin (PRF), an autogenous concentrated blood product, is a fibrin matrix comprising molecular and cellular elements that allow for optimum improvement and PRF also increases the production of osteoprotegerin, which causes osteoblast proliferation and acts as an osteoconductive and osteoinductive material, initiating bone regeneration. Platelet-rich plasma (PRP) and PRF are platelet concentrates prepared from the patient's own blood. Recent studies have focused on the development of therapeutic alternatives that are simple to prepare, non-toxic or biocompatible to living tissues, and inexpensive, with the potential to result in the local release of growth factors that accelerate hard and soft tissue healing. PRP is a plasma fraction that contains a high concentration of growth factors. PRF is a natural fibrin-based biomaterial derived from an anticoagulant-free blood harvest without any artificial biochemical modification, allowing fibrin membranes enriched with platelets and growth factors to be obtained. It facilitates the application of growth factors to the surgical area in a concentrated form, thereby accelerating wound improvement and regeneration. Platelet concentrates are used in many areas of dentistry, including soft tissue improvement, plastic periodontal surgery, gingival enlargement, ridge preservation, regeneration of bone defects, drug-related osteonecrosis of the jawbone, sinus augmentation, immediate implant placement, and implant osseointegration. Their use is preferred due to their ease of application and low cost. This paper reviews the current literature concerning the utilization of platelet-rich fibrin in periodontology. How to cite this article: Sağsöz A, Acun Kaya F. Use of platelet-rich fibrin (PRF) in periodontology: A review. J Med Dent Invest 2023;4:e230317. https://doi.org/10.5577/jomdi.e230317 Linguistic Revision: The English in this manuscript has been checked by at least two professional editors, both native speakers of English.

Local response and emerging nonlinear elastic length scale in biopolymer matrices

Nonlinear stiffening is a ubiquitous property of major types of biopolymers that make up the extracellular matrices (ECM) including collagen, fibrin, and basement membrane. Within the ECM, many types of cells such as fibroblasts and cancer cells have a spindle-like shape that acts like two equal and opposite force monopoles, which anisotropically stretch their surroundings and locally stiffen the matrix. Here, we first use optical tweezers to study the nonlinear force-displacement response to localized monopole forces. We then propose an effective-probe scaling argument that a local point force application can induce a stiffened region in the matrix, which can be characterized by a nonlinear length scale R* that increases with the increasing force magnitude; the local nonlinear force-displacement response is a result of the nonlinear growth of this effective probe that linearly deforms an increasing portion of the surrounding matrix. Furthermore, we show that this emerging nonlinear length scale R* can be observed around living cells and can be perturbed by varying matrix concentration or inhibiting cell contractility.

Comparative evaluation of the effectiveness of Titanium platelet rich fibrin (T-PRF) membrane and collagen membrane combined with xenograft in the treatment of mandibular class II furcation defects – A study protocol of randomized controlled trial

BACKGROUND Class II furcation is a significant clinical problem in dentistry. Its presence indicates advanced periodontitis. Different xenografts are available for the treatment of furcation involvement. Platelet concentrates that have been assimilated have a high concentration of growth factors, which has the added benefit of improving and accelerating healing. This study aims to compare the evaluation of effectiveness of T-PRF with xenograft and collagen membrane with xenograft in class II furcation defects. METHODS A one-year timeframe is going to be utilized to carry out the planned protocol, which is a randomised clinical trial. According to the inclusion criteria, 26 systemically healthy patients with class II furcation defects in mandibular molars will be included in the study. In Group I, xenograft and T-PRF (test group) will be condensed and in Group II, xenograft and collagen membrane (control group) will be condensed after the complete debridement of the furcation defect. Plaque index (PI), papillary bleeding index (PBI), Periodontal pocket depth (PPD), relative attachment level (R-CAL) and relative gingival marginal level (R-GML) will be evaluated as the clinical parameters. Cone-beam computed tomography (CBCT) will be performed to measure furcation height (FH), furcation width (FW), and furcation depth (FD) defects of mandibular molars. Data from each group will be evaluated at the time of surgery (baseline), and three and six months after the surgery. EXPECTED OUTCOMES Using xenograft (FIX OSS) combined with T-PRF, the authors anticipate better outcomes for class II mandibular furcation defects (test group). Improved clinical and radiographic outcomes in terms of PPD, R-CAL, R-GML and FH, FW, FD, are anticipated. CONCLUSIONS Using xenograft in combination with the T-PRF procedure is anticipated to have a better combined effect on periodontal parameters, including hard tissues and soft tissue outcomes. Registration: CTRI REF/2023/03/064367.

Use of platelet rich fibrin in tymapnoplasty

An observation study was undertaken at Department of Otorhinolaryngology, AIMSR fromJan 2021 – Aug 2022. The aim of my study the efficiency of use of autologous platelet rich fibrin membrane along with temporalis fascia in closure of tympanic membrane perforation during tympanopasty. All the patients attending ENT OPD at AIMSR within the age group of 11-60 years, irrespective of sex with tubotympanic CSOM either unilateral or bilateral were included in this study. In our study of 50 patients, 42 patients had graft uptake and 8 had failure of graft. Through this study we concluded that. The outcome of tympanoplasty does not depend on age and sex, dry ear of more than 6 month has a better graft.

The role of Platelet Rich Fibrin (PRF) membrane and conjunctival autograft on TGF-β and collagen type 1 expression in conjunctival wound healing: a literature review

Link of Video Abstract: https://youtu.be/8w4jBvx0pJc Background: Plasma-rich fibrine (PRF) membrane is a new biomembrane useful in ocular surface reconstruction technique. Since its development, PRF membrane has been used successfully in many studies; however, some controversies maintained. This literature study aims to evaluate the role of Platelet Rich Fibrin (PRF) membrane and conjunctival autograft on TGF-β and collagen type 1 expression in conjunctival wound healing. Methods: This literature review compiles and elaborates on previous studies from many authors to support future experimental studies, which will be conducted to compare the effect of PRF membrane and conjunctival autograft on TGF-β and type 1 collagen expression. Results: PRF has many advantages, such as the ability to reduce inflammation, rich in growth factors, and can be a temporary scaffold for fibroblast migration. However, despite its advantages, PRF is not widely used, unlike its counterpart techniques, such as conjunctival autograft. Conjunctival autograft has long been used as an option for ocular surface reconstruction techniques, but it has some limitations. The conjunctival autograft technique cannot be performed in large or bilateral conjunctival damage. Thus, the PRF membrane technique can become a perfect solution for such a case. This literature review will provide more information on how PRF can regulate inflammation and support the conjunctival wound healing process. Conclusion: Some impacts of PRF membrane and conjunctival autograft are still unclear, especially their effect on TGF-β and type 1 collagen expression. This literature review is expected to encourage and support future experimental studies about the impact of PRF and conjunctival autograft on TGF-β and type 1 collagen expression.

Management of Periapical cyst using Beta Tricalcium Phosphate Bone Substitute and Platelet Rich Fibrin Membrane Following Enucleation

The pulp and periodontium are connected anatomically, functionally, and developmentally. It might be challenging to diagnose and arrange effective treatments when pulpal issues and inflammatory periodontal disease coexist. One of the most frequent oral cavity cysts is the radicular cyst, which is typically brought on by trauma or an infection. There are a number of techniques that are now being used to enhance postoperative recovery and hasten the repair and regeneration of the damaged area. An investigation revealed that a 38-years-old patient had a radicular cyst, which was initially treated by endodontic treatment of the involved teeth, followed by enucleation of the cyst and a further bone graft placement followed by Platelet rich fibrin membrane. The patient's main complaint was pain in lower front jaw region for two to three months. Patient also had few episodes of pus discharge on its own through the same region. After three months, there was an increase in clinical attachment level and a decrease in probing depth. There was a sizable bone fill, according to radiographic findings.

Fat enhanced leukocyte-platelet-rich fibrin versus fascia lata in endoscopic reconstruction of CSF leaks

PurposeThe aim of this study was to use a new biological active fat enhanced leukocyte-platelet-rich fibrin membrane (L-PRF) for skull base defect reconstruction and compare its validity and reliability with the time-honored fascia lata.MethodsThis prospective study was conducted on 48 patients with spontaneous CSF leaks who were divided into 2 matched groups by stratified randomization, 24 patients in each group. In group A we performed multilayer repair using fat enhanced L-PRF membrane. In group B we used fascia lata for the multilayer repair. In both groups we enforced the repair with mucosal grafts/flaps.ResultsThe two groups were statistically matched for age, sex, intracranial pressure, and site and size of the skull base defect. There was no statistically significant difference between the two groups regarding the outcome of the repair or recurrence of CSF leak during the first postoperative year. Meningitis occurred in one patient in group B and was successfully treated. Another patient in group B developed thigh hematoma which resolved spontaneously.ConclusionThe fat enhanced L-PRF membrane is a valid reliable option in repair of CSF leaks. The membrane is autologous, readily available, easily prepared, and has the advange of including stromal fat, stromal vascular fraction (SVF), and leukocyte-platelet-rich fibrin (L-PRF). The present study showed that fat enhanced L-PRF membrane is stable, non-absorbable, not liable to shrink or become necrotic, and can establish good seal of the skull base defect and further enhance the healing process. The use of the membrane also has the advantage of avoiding thigh incision and possible hematoma formation.

Evaluation of PRF Membrane with Coronally Advanced Flap with and without Vertical Releasing Incisions for the Treatment of Gingival Recessions.

The aim of the current study was to compare the esthetic results for subjects with Miller Class I and II gingival recession (GR) abnormalities using platelet-rich fibrin (PRF) membrane with coronal advanced flaps (CAFs) with and without vertical releasing incisions (VRIs; the envelope-type flap and the flap with VRIs). Seven defects from each of the test and control groups made up of fourteen defects total. In the test group, PRF + CAF was performed without VRI, while in the control group, VRI was used. Gain in root coverage was the main result, with secondary results including papillary bleeding index (PBI), plaque index (PI), relative gingival margin level, relative attachment level, probing pocket depth, recession depth, width of keratinized gingiva (WKG), and gingival thickness. After 3 months of therapy, a clinical evaluation was conducted. No significant difference was observed between the two groups in terms of recession reduction (2.08 ± 0.5 vs 1.91 ± 0.66 mm), clinical attachment level (CAL) gain (2.08 ± 0.5 vs 1.91 ± 0.66 mm), and increase in WKG (2.66 ± 0.88 vs 2.58 ± 0.51 mm) for test and control groups, respectively. For the treatment of GR, both groups are efficient. However, the CAF + PRF without VRI group showed higher patient compliance and lower postoperative morbidity. The PRF membrane with CAF with or without VRI provide effective treatment option for GR. CAF + PRF without VRI is easy to perform and has less postoperative complications.

Clinical efficacy of titanium prepared platelet rich fibrin in periodontal regeneration: A systematic review and meta-analysis

Background: Periodontal regeneration therapies frequently involve autologous platelet concentrates (APCs). They can be used in sinus lift surgeries and socket preservation, among other clinical settings. Platelet rich fibrin (PRF) membrane has been used to treat gingival recession in individuals or groups of individuals using a coronally progressed or lateral pedicle flap. In the treatment of mixed periodontic endodontic lesion/furcation defect, PRF functions as a healing and interpositional biomaterial, filling a cystic cavity. PRF is known to help the bone regeneration process. In the last few years, efforts have been made to enhance the PRFs characteristics and quality. One of them is titanium platelet rich fibrin (T-PRF). Third-generation platelet concentrate no longer contains silica, and its preparation in glass vacuum containers, no longer creates any known concerns. The effectiveness PRF's has been evaluated in connective tissue and bone repair. The aim of this study is to compare T-PRF to other platelet concentrates and different treatment modalities for periodontal regenerative procedures. Methods: A protocol of this systematic review have been registered in prospero (CRD42022293545). The online database searched were PUBMED, COCHRANE for published articles up to November 2022 without language restrictions. Studies in trial registers, handsearching, bibliographic references of relevant articles were also checked. Data collection and analysis was done by individual authors. Independent eligibility assessments were conducted by four review authors. Then, using the standard Cochrane methodology, four review authors extracted the data and evaluated the risk of bias for individual studies. We developed "Summary of findings" tables and used GRADE to evaluate the evidence. Results: Three studies were included for meta-analysis. Results of meta-analysis supported that T-PRF is effective for correction of both hard and soft tissue defects. Conclusions: The overall qualitative and quantitative analysis suggest that T-PRF has superior structural properties and thicker fibrin network for periodontal regenerative procedures.

Comparative analysis of coronally advanced flap using platelet-rich fibrin membrane and amniotic membrane in gingival recession

Comparative analysis of coronally advanced flap using platelet-rich fibrin membrane and amniotic membrane in gingival recession

Comparative evaluation of the effectiveness of titanium platelet rich fibrin (T-PRF) membrane and collagen membrane combined with xenograft in the treatment of mandibular class II furcation defects –A Randomized Controlled Trial

Comparative evaluation of the effectiveness of titanium platelet rich fibrin (T-PRF) membrane and collagen membrane combined with xenograft in the treatment of mandibular class II furcation defects –A Randomized Controlled Trial

Comparative evaluation of the effectiveness of titanium platelet rich fibrin (T-PRF) membrane and collagen membrane combined with xenograft in the treatment of mandibular class II furcation defects –A Randomized Controlled Trial

Comparative evaluation of the effectiveness of titanium platelet rich fibrin (T-PRF) membrane and collagen membrane combined with xenograft in the treatment of mandibular class II furcation defects –A Randomized Controlled Trial

Impact of glutaraldehyde cross-linking on the advanced platelet-rich fibrin membrane: Macroscopic, conventional light microscopic and scanning electron microscopic analysis.

Advanced platelet-rich fibrin (A-PRF) is a biopolymer that releases growth factors to facilitate healing. Along with other barrier membranes, the A-PRF membrane has proven to be beneficial in guided tissue regeneration (GTR) treatment. The cross-linking of the A-PRF membrane with glutaraldehyde (GLUT) has been attempted previously, and has been shown to prolong its degradation time and improve its mechanical properties. In the present study, the effects of GLUT cross-linking on macroscopic changes in the A-PRF membrane were assessed, and microscopic features were analyzed using a light microscope and a scanning electron microscope (SEM). The aim of the present study was to evaluate and compare the effects of GLUT cross-linking on the A-PRF membrane through the macroscopic, microscopic and SEM examinations. A total of 18 human A-PRF membrane samples were prepared, half of which were treated with 0.1% GLUT, and the remaining were left untreated. The macroscopic measurements of the samples included weight, length and thickness, while specimen slides were prepared for light microscopic evaluation and SEM analysis. The GLUT cross-linked membranes weighed more and were thicker than the non-cross-linked membranes, but there was no change in length. Light microscopic images showed fewer cells at the head and tail, though cells were abundant in the body of the A-PRF membrane. The images acquired using SEM showed fibrin strands of greater thickness, but fewer interspersed cell bodies in the cross-linked membranes. This in vitro study revealed an increase in thickness and cross-linking fiber density along with the presence of viable cells in the GLUT-treated A-PRF membrane, which may prove its effectiveness in healing or serving as a barrier membrane in clinical trials.

Patient-reported outcome measures (PROMs) of leucocyte and platelet-rich fibrin (L-PRF) or hemostatic agent application at palatal donor sites after free gingival graft harvesting: a randomized controlled clinical trial.

The aim of this study was to evaluate the patient's morbidity and post-surgical complications after treatment of palatal donor sites after free gingival graft (FGG) harvesting using Leucocyte-and Platelet Rich Fibrin (L-PRF) membranes or a hemostatic agent with oxidized and regenerated cellulose. Forty-two palatal donar sites after FGG haversting in 42 patients were randomly assigned to experimental (L-PRF membrane) or control procedure (hemostatic agent). The primary outcome was postoperative pain related to the wound located at palatal area, while the secondary outcomes were post-operative discomfort, inability to chew, post-operative stress, surgical chair time, thickness of palatal fibromucosa and thickness of FGG. The PROMs were recorded after one week. After one week, a statistically significant difference was found between groups in terms of post operative stress (p=0.0083). No statistically significant differences in terms of postoperative pain (p=0.326), patient discomfort (p=0.509), inability to chew (p=0.936), surgical chair time (p=0.932) were recorded between test and control group. No statistically significant differences were recorded in terms of thickness of palatal fibromucosa (p=0.647) and thickness of FGG (P=0.756) between groups. Post-surgical wound healing complications (i.e., necrosis or infections) were not observed in both groups. Within their limitations, the present outcomes indicated that the application of L-PRF membrane at palatal donor sites after FGG harvesting did not produce significant advantages for the patients.

Clinical Evaluation of Coronally Advanced Flap With or Without Advance-Platelet Rich Fibrin Membrane in the Treatment of Miller's Class-II Localized Gingival Recession: A Clinical Study.

Periodontal treatment focuses on maintaining a patient's natural teeth and gums. The gingival margin recedes to a point apical to the tooth in 20%-100% of people. Coronallyadvanced flap (CAF) is one of several effective treatments for this condition. This surgery covers the tooth root with gingiva. The predictability of this surgery depends on the blood supply, donor tissue, and surgical skills. Platelet concentrates, which include platelet-rich fibrin (PRF), majority of the time is used for various regenerative therapies. Since no bovine thrombin or anticoagulant is needed, its manufacturing is simpler, cheaper, and less biochemically modified than PRP. Platelet-rich fibrin (PRF) is a fibrin matrix that progressively releases platelet cells and cytokines. The present study aimed to evaluate the efficacy of CAF with and without A-PRF in the treatment of Miller's class-II localized gingival recession. Twenty patients were chosen who had Miller's class-II localized gingival recession. A random number generator was used to place patients into either the "test" or "control" group. Treatment for both Groups A and B included a coronally advanced flap, but only Group A additionally got autologous platelet-rich fibrin (A-PRF). After receiving a detailed explanation of the treatment process, the patient signed an informed consent form. Complete medical and dental histories were taken to see whether there were any absolute or relative contraindications. Following treatment with either method in the current study, gingival thickness improved considerably. The percentage of root coverage did not change considerably between the two groups. The clinical result might likely have been different if other factors, such as platelet concentration and PRF consistency, had been examined in the current investigation. Furthermore, there was no histological examination of the healing process. As a result, we are unsure of the extent to which PRF affects how effectively connective tissue attaches. The additional use of A-PRF membrane did not provide additional benefits in terms of root coverage outcomes compared with CAF alone. The use of A-PRF membranes significantly reduced the recession depth.

Efficacy of leukocyte-platelet-rich fibrin membrane in immediate postextraction implant placement: A randomized controlled trial.

Leukocyte-platelet-rich fibrin (L-PRF) membrane is an emerging autologous healing biomaterial that promotes angiogenesis and healing in immediate implant sites. The purpose of the study was to evaluate hard and soft tissue outcomes of immediate implant placement with or without L-PRF. A total of 18 immediate implants were divided randomly into two groups of 9 implants each, i.e., Group 1 and Group 2. All sites received a definitive restoration after 3 months of implant placement and were followed up for a period of 6-months. Addition of L-PRF in the extraction sockets when placing immediate implants resulted in statistically nonsignificant benefit in terms of clinical and radiographic parameters when compared to immediate implant placement without L-PRF. Immediate implant placement in Group 2 demonstrated marginal but statistically significant benefit as compared to sites in Group 1.

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.