Dental extractions for patients on oral antiplatelet: a within-person randomised controlled trial comparing haemostatic plugs, advanced-platelet-rich fibrin (A-PRF+) plugs, leukocyte- and platelet-rich fibrin (L-PRF) plugs and suturing alone.
Giudice Amerigo,Esposito Marco,Bennardo Francesco,Brancaccio Ylenia,Buti Jacopo,Fortunato Leonzio
International journal of oral implantology (Berlin, Germany)
PURPOSE:To compare the outcome of tooth extractions in patients taking oral antithrombotic without reducing their dose. Four different interventions were compared within the same patient: suturing alone (control group), suturing plus a haemostatic plug, suturing plus advanced-platelet-rich fibrin (A-PRF+) plug, and suturing plus leukocyte- and platelet-rich fibrin (L-PRF) plug into the socket. MATERIALS AND METHODS:Forty patients, taking oral antiplatelet agents, requiring the extraction of at least four non-adjacent teeth were selected for the study. After extractions the sockets were randomly allocated to suturing alone (control group), suturing plus haemostatic plug (HAEM), suturing plus advanced-platelet-rich fibrin (A-PRF+) plug, and suturing plus leukocyte- and platelet-rich fibrin (L-PRF) plug into the socket without reducing the dose of oral antiplatelets according to a split-mouth design. Outcome measures were complications, time to complete each procedure, postoperative bleeding, costs of the materials, patient preference and a wound healing index recorded 1 and 2 weeks postextraction by blinded assessors. RESULTS:Two weeks after extraction no patient dropped out and no complication was reported. The average time to complete suturing after tooth extractions was: 1.0 ± 0.00 minutes at control sites, 1.5 ± 0.41 at HAEM sites, 2.8 ± 0.61 at A-PRF+ sites, and 2.8 ± 0.56 at L-PRF sites, the difference being statistically significant between each pairwise comparison except A-PRF+ vs L-PRF. Postoperative bleeding 30 minutes after extractions was present at 8, 5, 1 and 2 sites for control, HAEM, A-PRF+ and L-PRF sites, respectively. A-PRF showed statistically significantly less bleeding compared to the control group (odds ratio = 0.1 (95% CI [0.01;0.86]; P < 0.0361). In all cases bleeding was moderate in nature and not severe. One week after extractions the mean wound healing index was 1.05 ± 0.60 for control, 1.18 ± 0.59 for HAEM, 1.00 ± 0.68 for A-PRF+ and 0.95 ± 0.50 for L-PRF sites. No statistically significant difference was detected across groups (P = 0.633). Two weeks after extractions the mean wound healing index was 0.33 ± 0.53 for control, 0.43 ± 0.50 for HAEM, 0.25 ± 0.49 for A-PRF+ and 0.15 ± 0.36 for L-PRF sites. No statistically significant difference across groups was detected (P = 0.255). One week after extractions, nine patients preferred control sites, eight HAEM, ten A-PRF+, four L-PRF and nine had no preference. No statistically significant differences were detected for control sites (P = 0.6779), HAEM (P = 1.0000), A-PRF+ (P = 0.4055) and L-PRF (P = 0.1472). Two weeks after extractions five patients preferred control sites, three HAEM, eight A-PRF+, eight L-PRF and 16 had no preference. No statistically significant differences were detected for control sites (P = 0.8147), HAEM (P = 0.2363), A-PRF+ (P = 0.3488) and L-PRF (P = 0.3488). Costs without counting sutures and blood centrifuges were 0.00, 14.49, 2.44 and 2.44 Euro for control, HAEM, A-PRF+ and L-PRF sites, respectively. CONCLUSIONS:It may not be necessary to discontinue the use of oral antiplatelets in patients undergoing dental extractions and, when present, the minor statistically significant differences between procedures were not clinically relevant; therefore clinicians can use any of the tested interventions according to their preference, keeping in mind that simple suturing is sufficient and is faster and cheaper, and that A-PRF+ was associated with less postoperative bleeding when compared to suturing alone.
The impact of platelet-rich fibrin (PRF) on olfactory function and pain after septoplasty operations.
Tutar Belgin,Ekincioglu Enis,Karaketir Semih,Berkiten Güler,Saltürk Ziya,Arkan Melis Ece,Göker Ayşe Enise,Uyar Yavuz
European archives of oto-rhino-laryngology : official journal of the European Federation of Oto-Rhino-Laryngological Societies (EUFOS) : affiliated with the German Society for Oto-Rhino-Laryngology - Head and Neck Surgery
BACKGROUND:We aimed to investigate the effect of platelet-rich fibrin (PRF) on olfactory function and pain score in patients who underwent septoplasty. METHODS:This prospective randomized observational study was performed between 2018 January and 2019 April with 148 patients who had septoplasty operation. Patients were divided two groups and 74 patients were placed in group 1 to which PRF was applied after the completion of septoplasty whereas 67 patients were put in group 2 which did not undergo PRF. Sniffin' Sticks test was applied to all patients at pre-op, post-op 1-week, 6-week, and 6-month. Pain scores of patients were measured with visual analogue scale at 1 and 3 week. RESULTS:The distribution of patients according to pre-op olfactory function (normo-hypo-anosmia), there was no significant differences statistically (p > 0.05). When we compared the 1-week post-op results of Sniffin' Sticks test of patients, we found differences between the groups (p < 0.05). It was observed in the early postoperative period that according to the Sniffin' Sticks test scores, the results of the PRF group were better than those of the non-PRF group. At 6-week and 6-month, between the groups; there was no differences in terms of olfactory function. When we looked at the pain score of patients at 1 and 3 week after septoplasty; significant differences were obtained between groups. CONCLUSION:The application of PRF to the mucosal surface after the completion of septoplasty, has positive effect on olfactory function and pain especially in the early postoperative period. During the healing process, it was observed that prf maintained better odor functions. It is a minimally invasive technique with low risks and satisfactory clinical results.
10.1007/s00405-020-05839-6
The effect of age, gender, and time between blood draw and start of centrifugation on the size outcomes of platelet-rich fibrin (PRF) membranes.
Miron Richard J,Dham Anika,Dham Uttma,Zhang Yufeng,Pikos Michael A,Sculean Anton
Clinical oral investigations
OBJECTIVES:Platelet-rich fibrin (PRF) has been utilized in regenerative dentistry as a supra-physiological concentrate of autologous growth factors capable of stimulating tissue regeneration. Due to the variability in the macroscopic morphology/size of PRF membranes observed between patients, we were interested in studying the effects of patient age, gender, and time between blood draw and the start of centrifugation on the size outcomes of PRF membranes. Despite PRF therapy being increasingly more popular in private practice, to date, no study has investigated the effects of the delay between blood draw and the start of centrifugation in a clinical setting. MATERIALS AND METHODS:A total of 60 patients enrolled in this study were divided into 6 groups of 10 patients each, including male and female patients categorized into age groups 21-40, 41-60, and 61-80 years. From each patient, a total of five PRF membranes were fabricated from 10-mL tubes following centrifugation starting after 0, 30, 60, 90, and 120 s. In total, 300 PRF membranes were produced in this study to investigate the effects of patient age, gender, and time on the size outcomes of PRF membranes. RESULTS:A longer delay by the clinician before starting centrifugation following blood draw led to a smaller final size of PRF membranes. At 90 s following blood draw, a significant (13%) reduction in PRF membrane size was observed. After 120 s, a significant (23%) reduction was observed. Additionally, female patients had on average 17% larger membranes compared to men (p < 0.05, 300 samples). Lastly, the size outcomes of the PRF membranes was largest in patients aged 61-80, followed by those aged 41-60 and 21-40. However, no statistically significant differences in PRF membrane sizes were reported between age groups. CONCLUSIONS:The time at which a centrifugation procedure begins following blood draw is critical to optimize the size outcomes of PRF membranes. In general, approximately 15 s is required per tube to harvest 9-10 cc of blood. Therefore, a 60- to 90-s interval between blood draw and the start of centrifugation should be a parameter that is respected by clinicians to avoid significant changes in the macroscopic morphology/size of fabricated PRF membranes. Furthermore, females and older patients produced larger membranes, likely due to lower red blood cell counts derived from their peripheral blood. CLINICAL RELEVANCE:The findings from the present study demonstrate that on average, a clinician has approximately 60-90 s between blood draw and the start of the centrifugation cycle to produce standard-sized PRF membranes. Shortly thereafter, a significant reduction in size is observed. Additionally, females and older patients were found to produce larger PRF membranes. Centrifugation protocols may therefore be adapted accordingly.
10.1007/s00784-018-2673-x
Efficacy of PRF vs PRF + Biodegradable Collagen Plug in Post-extraction Preservation of Socket.
Ahmed Nida,Gopalakrishna Vivek,Shetty Akshay,Nagraj Vaibhav,Imran Mohammed,Kumar Praveen
The journal of contemporary dental practice
AIM:To compare the clinical sequelae of the efficacy of PRF vs PRF + collagen plug in soft tissue healing and preservation of the socket width, height, and bone density in patients reporting for extractions of maxillary or mandibular anterior or posterior teeth and patients who desired replacement of teeth with dental implants in future. MATERIALS AND METHODS:The study included 54 patients who were divided randomly into 3 groups consisting of 18 patients in each group: in group I, no preservation of extraction socket; in group II, PRF was used; and in group III, PRF + collagen plug was used for preservation of extraction socket. Assessment of the soft tissue healing, bone density, bone height, and width was done on 1st, 8th, 12th, and 16th weeks, postoperatively. RESULT:Both PRF and PRF + Collaplug are comparable to each other in preserving the bone height, bone density, and also similar soft tissue healing; however PRF + Collaplug is better than PRF alone in preserving the bone width 4th month postoperatively, indicating that the resorbable Collaplug® does play an additional role in preserving the socket width. CONCLUSION:PRF + Collaplug® has better clinical outcome in socket preservation in comparison to PRF alone. However, as results were not statistically significant, subjecting a larger sample size with PRF + Collaplug® for socket preservation may result in statistical critical values to substantiate our observations. CLINICAL SIGNIFICANCE:PRF and Collaplug® can help in ridge preservation after extraction and also avoid additional bone grafting procedures in future implant placement for the patients. How to cite this article: Ahmed N, Gopalakrishna V, Shetty A, Efficacy of PRF vs PRF + Biodegradable Collagen Plug in Post-extraction Preservation of Socket. J Contemp Dent Pract 2019;20(11):1323-1328.
PRF improves wound healing and postoperative discomfort after harvesting subepithelial connective tissue graft from palate: a randomized controlled trial.
Lektemur Alpan Aysan,Torumtay Cin Gizem
Clinical oral investigations
OBJECTIVES:The aim of this study is to determine the use of platelet-rich fibrin (PRF) in the management of soft tissue donor site healing after harvesting connective tissue graft (CTG) from the palate and evaluate the postoperative discomfort (pain, bleeding, analgesic consumption, tissue color match) of patients. MATERIALS AND METHODS:Forty patients were randomly assigned to PRF or control group. In the PRF group, PRF membrane was placed into CTG donor site. After surgery, delayed bleeding, early healing index (EHI), tissue color match, and analgesic consumption were recorded. The visual analog scale (VAS) was used to observe the postoperative pain and tissue color match. Data were analyzed using the independent sample t test and the repeated measure ANOVA test. RESULTS:The patients in the PRF group reported significantly lower pain scores at all-time points. Postoperative 3rd and 7th day, EHI scores were lower in the favor of the PRF group. VAS score values of tissue color match were lower in the control group at 7th and 14th day, compared with the PRF group. Analgesic intake was significantly lower in the PRF group postoperatively at 1st and 3rd day that of control group. CONCLUSION:PRF application at the palatal donor site demonstrates promising results in terms of better wound healing and reduced postoperative discomfort in the patients after harvesting CTG. CLINICAL RELEVANCE:Patients may avoid surgical operations because of the discomfort or pain feeling. Reducing postoperative pain and discomfort and accelerating recovery meet the wishes of every patient and physician. PRF can provide these requirements as an easy method to obtain and implement.
10.1007/s00784-019-02934-9
Biological characterization of an injectable platelet-rich fibrin mixture consisting of autologous albumin gel and liquid platelet-rich fibrin (Alb-PRF).
Fujioka-Kobayashi Masako,Schaller Benoit,Mourão Carlos Fernando De Almeida Barros,Zhang Yufeng,Sculean Anton,Miron Richard J
Platelets
Platelet-rich fibrin (PRF) has been proposed as an autologous membrane with the advantages of host accumulation of platelets and leukocytes with entrapment of growth factors. However, limitations include its faster resorption properties (~2 weeks). Interestingly, recent studies have demonstrated that by heating a liquid platelet-poor plasma (PPP) layer, the resorption properties of heated albumin (albumin gel) can be extended from 2 weeks to greater than 4 months (e-PRF). The aim of the present study was to characterize the biological properties of this novel regenerative modality. Whole blood collected from peripheral blood in 9-mL plastic tubes was centrifuged at 700 for 8 minutes. Thereafter, the platelet-poor plasma layer was heated at 75°C for 10 minutes to create denatured albumin (albumin gel). The remaining cells and growth factor found within the buffy coat layer (liquid PRF) were thereafter mixed back together with the cooled albumin gel to form Alb-PRF. Histological analysis, including the distribution of cells within Alb-PRF, was then performed. Seven different growth factor release kinetics from Alb-PRF were characterized up to 10 days, including PDGF-AA, PDGF-AB, PDGF-BB, TGF-β1, VEGF, IGF and EGF. Thereafter, gingival fibroblast cell responses to Alb-PRF were investigated by means of a live/dead assay at 24 hours; migration assay at 24 hours; proliferation assay at 1, 3 and 5 days; real-time PCR for the expression of TGF-β and collagen 1a2 at 3 and 7 days; and collagen 1 immunostaining at 14 days. It was first observed histologically that viable cells were evenly distributed throughout the Alb-PRF formulation. Growth factor release demonstrated a slow and gradual release, particularly for TGF-β1 and PDGF-AA/AB, during the entire 10-day period. Alb-PRF also exhibited statistically significantly higher cell biocompatibility at 24 hours and statistically significantly induced greater fibroblast proliferation at 5 days when compared to those of control TCP. Alb-PRF further induced statistically significantly greater mRNA levels of TGF-β at 3 and 7 days, as well as collagen 1 at 7 days. The present results indicate that Alb-PRF possesses regenerative properties induced by the slow and gradual release of growth factors found in liquid PRF via albumin gel degradation. Future studies are thus warranted to fully characterize the degradation properties of Alb-PRF and explore future clinical applications in various fields of medicine.
10.1080/09537104.2020.1717455
Impact of incubation method on the release of growth factors in non-Ca-activated PRP, Ca-activated PRP, PRF and A-PRF.
Steller Daniel,Herbst Nele,Pries Ralph,Juhl David,Hakim Samer G
Journal of cranio-maxillo-facial surgery : official publication of the European Association for Cranio-Maxillo-Facial Surgery
The aim of this study was to investigate the influence of different incubation methods on the growth factor content of lysates of platelet-rich fibrin (PRF), advanced-platelet-rich fibrin (A-PRF) and platelet-rich plasma (PRP) products. A comparison of related studies suggests that the method of sample preparation has a significant influence on growth factor content. There are few reports on the comparison of non-Ca-activated PRP, Ca-activated PRP, A-PRF, and PRF, along with a lack of information on the release of PDGF-BB, TGF-β1, and VEGF among the different incubation methods. The lysate preparation was made of non-Ca-activated PRP, Ca-activated PRP, PRF, and A-PRF, using a room-temperature, 37 °C, or freeze-thaw-freeze incubation method. Afterwards the VEGF, PDGF-BB, and TGF-β1 content was investigated by running ELISA tests. Growth factor levels were significantly increased in the non-Ca-activated PRP with freeze-thaw-freeze incubation, and in the PRF preparation there was a significant disadvantage to using room temperature incubation for releasing growth factors. In conclusion, the freeze-thaw-freeze method is sufficient for releasing growth factors, and calcium activation is not necessary. Finally, the study demonstrates the possibility of preparing PRP products from platelet concentrates, so that preoperative blood sampling might not be required.
10.1016/j.jcms.2018.10.017
The Effects of Leukocyte-Platelet Rich Fibrin (L-PRF) on Suppression of the Expressions of the Pro-Inflammatory Cytokines, and Proliferation of Schwann Cell, and Neurotrophic Factors.
Wang Zhanqi,Mudalal Mahmoud,Sun Yue,Liu Yiping,Wang Jia,Wang Yao,Sun Xiaolin,Zhou Yanmin
Scientific reports
This study evaluates the use of L-PRF as an autologous scaffold in nerve regeneration, and Schwann cells (SCs) proliferation and secretion of neurotrophic factors and its anti-inflammatory effect on SC Porphyromonas Gingivalis-Lipopolysaccharide (PG-LPS)-induced inflammatory responses in vitro. SEM was done to investigate various features of L-PRF. L-PRF-extracts was used to investigate the release of growth factors and treatment of SCs line. ELISA was applied to examine the release of IGF-1. The proliferative effect of L-PRF on SCs was assessed with CCK-8 assay. The effect of L-PRF on the mRNA and protein expression of SC neurotrophic factors were analyzed by RT-qPCR and ELISA. CCK-8 assay and RT-qPCR were used to determine the required concentration and the action time of PG-LPS before the anti-inflammatory effect of L-PRF was determined by measuring the changes in IL-1β, IL-6, and TNF-a with RT-qPCR and ELISA. There are different features in L-PRF. Fourteen days was sufficient to release adequate GF. The mRNA expressions of the pro-inflammatory cytokines were notably raised by PG-LPS in 3-hours treatment. L-PRF can increase SC proliferation, neurotrophic factors secretion, and suppress SC PG-LPS-induced inflammatory responses in vitro. L-PRF has the potential as an autologous biological additive for peripheral nerve regeneration in the event of nerve inflammation and injuries.
10.1038/s41598-020-59319-2
Comparison of platelet-rich fibrin (PRF) produced using 3 commercially available centrifuges at both high (~ 700 g) and low (~ 200 g) relative centrifugation forces.
Miron Richard J,Xu Hudi,Chai Jihua,Wang Jiaolong,Zheng Shihang,Feng Mengge,Zhang Xiaoxin,Wei Yan,Chen Yan,Mourão Carlos Fernando de Almeida Barros,Sculean Anton,Zhang Yufeng
Clinical oral investigations
OBJECTIVES:Platelet-rich fibrin (PRF) has gained tremendous momentum in recent years as a natural autologous growth factor derived from blood capable of stimulating tissue regeneration. Owing to its widespread use, many companies have commercialized various centrifugation devices with various proposed protocols. The aim of the present study was to compare 3 different commercially available centrifuges at both high and low g-force protocols. MATERIALS AND METHODS:PRF was produced on three commercially available centrifuges including the IntraSpin Device (IntraLock), the Duo Quattro (Process for PRF), and Salvin (Salvin Dental). Two separate protocols were tested on each machine including the original leukocyte and platelet-rich fibrin (L-PRF) protocol (~ 700 RCF max (~ 400 RCF clot) for 12 min) as well as the advanced platelet-rich fibrin (A-PRF+) protocol (~ 200 g RCF max (~ 130 g RCF clot) for 8 min). Each of the tested groups was compared for cell numbers, growth factor release, scanning electron microscopy (SEM) for morphological differences, and clot size (both weight and length/width). RESULTS:The present study found that PRF clots produced utilizing the low-speed centrifugation speeds (~ 200 g for 8 min) produce clots that (1) contained a higher concentration of evenly distributed platelets, (2) secreted higher concentrations of growth factors over a 10 day period, and (3) were smaller in size. This was irrespective of the centrifugation device utilized and consistently observed on all 3 devices. The greatest impact was found between the protocols utilized (up to a 200%). Interestingly, it was further revealed that the centrifugation tubes used had a much greater impact on the final size outcome of PRF clots when compared to centrifugation devices. It was found that, in general, the Process for PRF tubes produced significantly greater-sized clots when compared to other commercially available tubes. The Salvin Dental tubes also produced significantly greater PRF clots when compared to the IntraLock tubes on each of the tested centrifugation devices. CONCLUSIONS:The present study demonstrated the reproducibility of a scientific concept (reduction in RCF produces PRF clots with more evenly distributed cells and growth factors) utilizing different devices. Furthermore, (and until now overlooked), it was revealed for the first time that the centrifugation tubes are central to the quality production of PRF. Future research investigating tube characteristics thus becomes critically important for the future optimization of PRF. CLINICAL RELEVANCE:This is the first study to reveal the marked impact of centrifugation tubes on the final production of PRF. Future study thus becomes markedly important to further optimize the quality of PRF-based matrices. It was further found that little variability existed between the centrifugation devices if optimized centrifugation protocols (lower centrifugation speeds) were utilized.
10.1007/s00784-019-02981-2
Improved growth factor delivery and cellular activity using concentrated platelet-rich fibrin (C-PRF) when compared with traditional injectable (i-PRF) protocols.
Fujioka-Kobayashi Masako,Katagiri Hiroki,Kono Michihide,Schaller Benoit,Zhang Yufeng,Sculean Anton,Miron Richard J
Clinical oral investigations
OBJECTIVES:Several studies have recently demonstrated that only marginal improvements in platelet and leukocyte concentrations are achieved following standard injectable platelet-rich fibrin (i-PRF) protocols. Due to these previous findings, a novel harvesting technique was recently developed to collect higher concentrations of platelets/leukocytes specifically from the buffy coat layer (C-PRF) following faster centrifugation protocols. The aim of this study was to investigate the regenerative properties and effects on growth factor release and cellular activity of PRF collected through this novel harvesting technique compared to standard i-PRF protocols. MATERIALS AND METHODS:The upper 1-ml layer collected through standard i-PRF protocols at low centrifugation speeds was compared with 1 mL of C-PRF collected from the buffy coat layer following high centrifugation protocols (3000×g for 8 min on a horizontal centrifuge) to specifically concentrate cells within the platelet/leukocyte-rich buffy coat layer. Thereafter, the expression of seven different growth factors, including PDGF-AA, PDGF-AB, PDGF-BB, TGF-β1, VEGF, IGF-1, and EGF, was characterized for up to 10 days. Then, gingival fibroblast biocompatibility was investigated at 24 h (live/dead assay); migration was investigated at 24 h; proliferation was investigated at 1, 3, and 5 days; and the expression of PDGF and TGF-β was investigated at 3 days. Collagen 1 immunostaining was also quantified at 14 days. RESULTS:At all investigated time periods, a significant increase in growth factor release was observed in C-PRF. In particular, the release of PDGF-AA, TGF-β1, and EGF exhibited the highest increases when compared with that in i-PRF. While both i-PRF and C-PRF exhibited high biocompatibility and induced significantly higher fibroblast migration and proliferation when compared with that of the control tissue culture plastic group, C-PRF showed the greatest potential for cell migration and proliferation. Furthermore, C-PRF induced significantly higher mRNA levels of TGF-β and PDGF levels at 3 days and greater collagen 1 staining when compared with induced by i-PRF. CONCLUSIONS:In the present study, it was found that C-PRF collected specifically from the buffy coat layer following higher centrifugation protocols exhibited an up to a threefold increase in growth factor release when compared with that exhibited by standard i-PRF. This significantly promoted higher gingival fibroblast migration, proliferation, gene expression, and collagen I synthesis. CLINICAL RELEVANCE:The findings of the present study demonstrate that a more potent formulation of liquid platelet concentrate than that obtained from the upper plasma layer following a short and slow centrifugation protocol (i-PRF protocol) can be obtained for clinical use by specifically harvesting cells in the platelet- and leukocyte-rich buffy coat layer following an 8-min 3000×g centrifugation protocol (C-PRF protocol).
10.1007/s00784-020-03303-7
Quantitative Near-Infrared Imaging of Platelets in Platelet-Rich Fibrin (PRF) Matrices: Comparative Analysis of Bio-PRF, Leukocyte-Rich PRF, Advanced-PRF and Concentrated Growth Factors.
Aizawa Hachidai,Tsujino Tetsuhiro,Watanabe Taisuke,Isobe Kazushige,Kitamura Yutaka,Sato Atsushi,Yamaguchi Sadahiro,Okudera Hajime,Okuda Kazuhiro,Kawase Tomoyuki
International journal of molecular sciences
Platelet-rich fibrin (PRF) is a fibrin matrix enriched with platelets. The PRF matrix is thought to form a steep gradient of platelet density around the region corresponding to the buffy coat in anticoagulated blood samples. However, this phenomenon has not yet been proven. To visualize platelet distribution in PRF in a non-invasive manner, we utilized near-infrared (NIR) imaging technology. In this study, four types of PRF matrices, bio-PRF, advanced-PRF (A-PRF), leukocyte-rich PRF (L-PRF), and concentrated growth factors (CGF) were compared. Blood samples collected from healthy, non-smoking volunteers were immediately centrifuged using four different protocols in glass tubes. The fixed PRF matrices were sagittally divided into two equal parts, and subjected to modified immunohistochemical examination. After probing with NIR dye-conjugated secondary antibody, the CD41 platelets were visualized using an NIR imager. In L-PRF and CGF, platelets were distributed mainly on and below the distal surface, while in bio-PRF and A-PRF, platelet distribution was widespread and homogenous. Among three regions of the PRF matrices (upper, middle, and lower), no significant differences were observed. These findings suggest that platelets aggregate on polymerizing fibrin fibers and float up as a PRF matrix into the plasma fraction, amending the current "gradient" theory of platelet distribution.
10.3390/ijms21124426
A novel method for harvesting concentrated platelet-rich fibrin (C-PRF) with a 10-fold increase in platelet and leukocyte yields.
Miron Richard J,Chai Jihua,Zhang Peng,Li Yuqing,Wang Yunxiao,Mourão Carlos Fernando de Almeida Barros,Sculean Anton,Fujioka Kobayashi Masako,Zhang Yufeng
Clinical oral investigations
BACKGROUND AND OBJECTIVES:Liquid platelet rich fibrin (PRF; often referred to as injectable PRF) has been utilized as an injectable formulation of PRF that is capable of stimulating tissue regeneration. Our research group recently found that following standard L-PRF protocols (2700 RPM for 12 min), a massive increase in platelets and leukocytes was observed directly within the buffy-coat layer directly above the red blood cell layer. The purpose of this study was to develop a novel harvesting technique to isolate liquid PRF directly from this buffy coat layer and to compare this technique to standard i-PRF. MATERIALS AND METHODS:Standard high g-force L-PRF and low g-force i-PRF protocols were utilized to separate blood layers. Above the red blood corpuscle layer, sequential 100-μL layers of plasma were harvested (12 layers total; i.e., 1.2 mL, which represents the total i-PRF volume), and 3 layers (3 × 100 μL) were harvested from the red blood cell layer to quantify blood cells. Each layer was then sent for complete blood count (CBC) analysis, and the cell numbers were quantified including red blood cells, leukocytes, neutrophils, lymphocytes, monocytes, and platelets. The liquid PRF that was directly collected from the buffy-coat layer following L-PRF protocols was referred to as concentrated PRF (C-PRF). RESULTS:The i-PRF protocol typically yielded a 2- to 3-fold increase in platelets and a l.5-fold increase in leukocyte concentration from the 1- to 1.2-mL plasma layer compared to baseline concentrations in whole blood. While almost no cells were found in the first 4-mL layer of L-PRF, a massive accumulation of platelets and leukocytes was found directly within the buffy coat layer demonstrating extremely high concentrations of cells in this 0.3-0.5-mL layer (~ 20-fold increases). We therefore proposed harvesting this 0.3- to 0.5-mL layer directly above the red blood cell corpuscle layer as liquid C-PRF. In general, i-PRF was able to increase platelet numbers by ~ 250%, whereas a 1200-1700% increase in platelet numbers could easily be achieved by harvesting this 0.3-0.5 mL of C-PRF (total platelet concentrations of > 2000-3000 × 10 cells/L). CONCLUSION:While conventional i-PRF protocols increase platelet yield by 2-3-fold and leukocyte yield by 50%, we convincingly demonstrated the ability to concentrate platelets and leukocytes over 10-fold by harvesting the 0.3-0.5 mL of C-PRF within the buffy coat following L-PRF protocols. CLINICAL RELEVANCE:Previous studies have demonstrated only a slight increase in platelet and leukocyte concentrations in i-PRF. The present study described a novel harvesting technique with over a 10-fold increase in platelets and leukocytes that can be further utilized for tissue regeneration.
10.1007/s00784-019-03147-w
Advanced-PRF: Clinical evaluation in impacted mandibular third molar sockets.
Gupta N,Agarwal S
Journal of stomatology, oral and maxillofacial surgery
OBJECTIVE:This prospective study evaluated the efficacy and healing potential of modified formulation of PRF, commonly known as Advanced PRF (A-PRF) in impacted mandibular third molar extraction sockets. MATERIALS AND METHODS:20 patients with bilateral impacted mandibular third molars were included in this study. Surgical disimpactions were done at 3-4 weeks interval in opposing quadrants of patient. One quadrant received A-PRF while the opposing quadrant in same patient was taken as control. Comparative evaluation was done in terms of pain assessment, analgesics required, swelling, soft tissue healing and trismus on 1st, 3 and 7day follow-up. Comparative assessment of bone healing was also done on 1st, 3and 6month follow-up. RESULTS:This study involved 12 female and 8 male patients with in age range of 18-35 years. The evaluation of pain, swelling, trismus and soft tissue healing on 3rd postoperative day revealed considerable improvement on A-PRF sites as compared to control sites. The outcomes were found to be statistically significant with p values 0.008, 0.031, 0.0001, 0.05 respectively. Even the analgesic consumption was remarkably less when A-PRF was used (P=0.004). Bone healing evaluation in A-PRF sites on 1st, 3and 6month has shown significantly improved results with P<0.05. CONCLUSION:Our study infers that A-PRF has enhanced the healing potential of soft tissue as well as bone in extraction socket. Apart from that it has also shown promising results in relief of immediate postoperative symptoms like pain, swelling and trismus which improves the comfort and acceptability of surgical procedures by patients. Enhanced healing and patient comfort in cost effective manner are the highlighting features of A-PRF.
10.1016/j.jormas.2020.04.008
Improvement of bone repair with l-PRF and bovine bone in calvaria of rats. histometric and immunohistochemical study.
do Lago Eliel Scarpioni,Ferreira Sabrina,Garcia Idelmo Rangel,Okamoto Roberta,Mariano Ronaldo Célio
Clinical oral investigations
OBJECTIVES:The effect of leucocyte- and platelet-rich fibrin (L-PRF), associated with DBBM (deproteinized bovine bone mineral; Bio-Oss®) was investigated and compared with autogenous bone graft as a standard material for filling bone defects. MATERIAL AND METHODS:A defect of 5 mm in diameter was performed in 40 calvaria of rats. The animals were divided into 5 groups and received blood clot (CO), autogenous bone (AUT), DBBM (BIO), L-PRF, or DBBM associated with L-PRF (BIO-LPRF). After 4 and 8 weeks, bone regeneration was assessed by histometric and immunohistochemical analyses. RESULTS:The highest mean percentage of bone formation found at 4 and 8 weeks was observed for the BIO-L-PRF group (54.0% ± 2.8 and 63.6% ± 2.2). The lowest mean percentage at 4 and 8 weeks was observed for the CO group (16.7% ± 2.5 and 20.5% ± 1.0). There was statistical similarity among the AUT, BIO, and L-PRF groups. The expressions OC, RUNX 2, and VEGF showed a favorable aspect in the formation of new bone for BIO-L-PRF. VEGF was the marker with the highest expression because it was related to the initial healing process, promoting the migration and proliferation of endothelial cells in the region of the defect. Even after weeks, VEGF maintained a moderate expression. CONCLUSIONS:The association of L-PRF with DBBM improved bone repair when these biomaterials were inserted into the defects of the calvaria of rats. CLINICAL RELEVANCE:This reinforces the good performance of bovine bone and L-PRF as filler materials, especially when associated.
10.1007/s00784-019-03018-4
In vivo evaluation of the biocompatibility and biodegradation of a new denatured plasma membrane combined with liquid PRF (Alb-PRF).
Gheno Ezio,Mourão Carlos Fernando de Almeida Barros,Mello-Machado Rafael Coutinho de,Stellet Lourenço Emanuele,Miron Richard J,Catarino Karoline Ferreira Farias,Alves Adriana Terezinha,Alves Gutemberg Gomes,Calasans-Maia Mônica D
Platelets
Guided bone regeneration (GBR) is a process that involves the regeneration of bone defects through the application of occlusive membranes that mechanically exclude the population of non-osteogenic cells from the surrounding soft tissue. Interestingly, platelet-rich fibrin (PRF) has previously been proposed as an autologous GBR membrane despite its short-term resorption period of 2-3 weeks. Recent clinical observations have demonstrated that, by heating a liquid platelet-poor plasma (PPP) layer and mixing the cell-rich buffy coat zone, the resorption properties of heated albumin gel with liquid-PRF (Alb-PRF) can be significantly improved. The aim of this study was to evaluate the inflammatory reaction, biocompatibility, and extended degradation properties of a new autologous Alb-PRF membrane in comparison to commonly utilized standard PRF after nude mice implantation, according to ISO 10993-6/2016. Two standard preparations of PRF (L-PRF and H-PRF) were compared to novel Alb-PRF following subcutaneous implantation at 7, 14, and 21 days. All groups demonstrated excellent biocompatibility owing to their autologous sources. However, it is worth noting that, while both L-PRF and H-PRF membranes demonstrated significant or complete resorption by 21 days, the Alb-PRF membrane remained volume-stable throughout the duration of the study. This study demonstrates-for the first time, to the best of our knowledge-a marked improvement in the membrane stability of Alb-PRF. This indicates its future potential for use as a biological barrier membrane for GBR procedures with a long-lasting half-life, or as a biological filler material in esthetic medicine applications. Thus, further studies are warranted to explore future clinical applications in various fields of medicine.
10.1080/09537104.2020.1775188