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PurposeThe aim of this study was to analyze systematically the influence of the relative centrifugation force (RCF) on leukocytes, platelets and growth factor release within fluid platelet-rich fibrin matrices (PRF).Materials and methodsSystematically using peripheral blood from six healthy volunteers, the RCF was reduced four times for each of the three experimental protocols (I–III) within the spectrum (710–44 g), while maintaining a constant centrifugation time. Flow cytometry was applied to determine the platelets and leukocyte number. The growth factor concentration was quantified 1 and 24 h after clotting using ELISA.ResultsReducing RCF in accordance with protocol-II (177 g) led to a significantly higher platelets and leukocytes numbers compared to protocol-I (710 g). Protocol-III (44 g) showed a highly significant increase of leukocytes and platelets number in comparison to -I and -II. The growth factors’ concentration of VEGF and TGF-β1 was significantly higher in protocol-II compared to -I, whereas protocol-III exhibited significantly higher growth factor concentration compared to protocols-I and -II. These findings were observed among 1 and 24 h after clotting, as well as the accumulated growth factor concentration over 24 h.DiscussionBased on the results, it has been demonstrated that it is possible to enrich PRF-based fluid matrices with leukocytes, platelets and growth factors by means of a single alteration of the centrifugation settings within the clinical routine.ConclusionsWe postulate that the so-called low speed centrifugation concept (LSCC) selectively enriches leukocytes, platelets and growth factors within fluid PRF-based matrices. Further studies are needed to evaluate the effect of cell and growth factor enrichment on wound healing and tissue regeneration while comparing blood concentrates gained by high and low RCF.

This study compared the efficacy of apicoectomy and apical curettage with and without leukocyte- and platelet-rich fibrin (L-PRF) in treating large periapical lesions. Sixty-four adults (30 male, 34 female) with a previously endodontically treated tooth and a large periapical lesion were randomized into four groups (n = 16): retrograde obturation (RG), orthograde obturation (OG), RG + L-PRF, and OG + L-PRF. All participants underwent root canal retreatment in two sessions. The RG group underwent root-end resection and retrograde MTA obturation, the RG + L-PRF group underwent L-PRF application to the bone defect following the RG protocol, the OG group underwent orthograde MTA obturation and periapical curettage without root-end resection, and the OG + L-PRF group underwent L-PRF application following the OG protocol. Clinical and radiographic assessments were performed preoperatively, and at 1 week and 1, 3, 6, 9, and 12 months postoperatively. At follow-up visits, pain scores, swelling, tooth mobility, tenderness to percussion (T-PER), tenderness to palpation (T-PAL), and the presence of fistula were clinically assessed. Periapical radiography determined the periapical index (PAI) score and measured the periapical lesion area (PALA). The Kruskal–Wallis test was performed to test the effect of a single independent variable (factor) on a dependent variable. No statistically significant differences were identified between the groups for preoperative PAI scores, pain scores, swelling, tooth mobility, fistula, T-PER, or T-PAL (p > 0.05). At postoperative week 1, the RG + L-PRF group showed a significantly lower T-PER. The RG + L-PRF group showed significantly lower PALA values and significantly higher PALA healing rates at postoperative 1, 6, and 9 months. Both L-PRF groups achieved PALA healing rates of over 90% at 9 months. It was concluded that a combination of apicoectomy and L-PRF effectively treats periapical lesions and promotes both short- and long-term healing and that a combination of periapical curettage and L-PRF offers a less invasive alternative, especially when the crown-to-root ratio is a concern. Trial registration: The protocol was registered at ClinicalTrials.gov (NCT05847647).

Platelet-rich fibrin (PRF) has emerged as a promising scaffold for drug delivery, particularly in the context of antimicrobial therapies. This systematic review evaluates the incorporation of antibiotics into PRF to determine its efficacy as a localized antimicrobial delivery system compared to plain PRF without antibiotics. A systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, including 13 in vitro studies with a moderate risk of bias. Antibiotics were incorporated into PRF using different methodologies, including systemic administration before blood collection, addition to blood before centrifugation, and injection into formed PRF matrices. Outcomes were analyzed regarding antibacterial efficacy, structural integrity of PRF, and release kinetics. Antibiotic-enhanced PRF demonstrated significant antibacterial activity against various bacterial strains. The efficacy of the enhanced PRF was dependent on the type of antibiotic, its concentration, and incorporation method. Encapsulation approaches facilitated a sustained antibiotic release, while higher antibiotic concentrations occasionally disrupted PRF integrity. Systemic administration of antibiotics before blood collection enriches PRF effectively, producing significant inhibition zones. The antibacterial effects of PRF outperformed alternative carriers, such as collagen sponges. Antibiotic-loaded PRF is a potent tool for localized antimicrobial delivery, with promising applications in clinical settings. Further research is needed to standardize preparation protocols and explore the impact of different antibiotic delivery methods on PRF’s regenerative properties.

ObjectivesThe aim of the present study was to evaluate the blood cell content, morphological aspects, gene expression of type I collagen, and release of growth factors on an injectable platelet rich fibrin (i-PRF).Materials and methodsBlood samples were collected from 15 volunteers to prepare i-PRF samples. Peripheral blood was used as a control group. Blood clot and i-PRF samples were cultured for 10 days. The supernatant of the samples was collected for ELISA immunoassay quantification of PDGF and VEGF growth factors over periods of 1, 8, 24, 72, and 240 h. I-PRF and blood clot samples were biologically characterized using histological and immunohistochemistry analysis for IL-10, osteocalcin, and TGF-β. Scanning electron microscopy (SEM) was used to inspect the fibrin network and distribution of blood platelets and leukocytes. Reverse transcriptase polymerase chain reaction (RT-PCR) method was used to evaluate gene expression for type I collagen.ResultsA higher concentration of platelets and lymphocytes was recorded in i-PRF than in peripheral blood (p < 0.05). The release of VEGF was higher in blood clot samples (1933 ± 704) than that for i-PRF (852 ± 376; p < 0.001). Immunohistochemistry showed upregulation of TGF-B, IL-10, and osteocalcin in the i-PRF group. RT-PCR showed increased type I collagen gene expression in i-PRF (p < 0.05). SEM images revealed agglomeration of platelets in some regions, while a fibrin networking was noticeable in the entire i-PRF sample.ConclusionsInjectable platelet rich fibrin becomes a good approach for soft and mineralized tissue healing considering the formation of a three-dimensional fibrin network embedding platelets, leukocytes, type I collagen, osteocalcin, and growth factors. Indeed, the injectable platelet rich fibrin can be indicated in several medical applications regarding bioactivity, simplied technique, and flowable mixing with other biomaterials.Clinical relevanceMorphological, cell, and protein characterization of platelet rich fibrin provides a better understanding of the clinical effects and improvement of clinical guidelines for several medical applications. Once well physicochemical and biologically characterized, the use of an injectable platelet rich fibrin can be extended to other applications in the field of orthopedics, periodontics, and implant dentistry on the repairing process of both soft and mineralized tissues.

Background The aim of this study was to evaluate 24 protocols for the production of platelet rich fibrin (PRF) produced via horizontal centrifugation to better understand cell separation following protocols at various times and speeds. Methods All protocols were compared utilizing a recent method to quantify cells in PRF in 1 mL sequential layers pipetted from the upper layer downwards until all 10 mL were harvested. In total, 960 complete blood counts (CBCs) were investigated. Both solid and liquid-based PRF protocols were investigated following 24 protocols involving 6 relative centrifugal force (RCF) values (100, 200, 400, 700, 1000 and 1200 g ) at 4 centrifugation times (3, 5, 8 and 12 min). Results In general, platelets could more easily accumulate in the upper 4 layers when compared to leukocytes owing to their lower cellular density. Protocol time seemed to have a greater impact on the final cell layer separation when compared to the effect of speed. Protocols of greater than 8 min at 400 g led to no leukocyte accumulation in the upper PRF layers (found specifically within the buffy coat). Protocols at or below 200 g were unable to effectively accumulate platelets/leukocytes. The optimal centrifugation speed and time for solid-PRF ranged between 400 and 700 g for 8 min. It was noted that variability in patient baseline platelet/leukocyte/erythrocyte counts (hematocrit) significantly affected cell layer separation. This finding was more pronounced at lower centrifugation speeds. Conclusions Within the investigated ranges, a protocol of 700 g for 8 min presented the highest yield of platelets/leukocytes evenly distributed throughout the upper PRF layers.

This study aimed to comparatively assess the clinical success and radiographic regenerative dentin formation of Platelet Rich Fibrin (PRF) and mineral trioxide aggregate (MTA) when used as direct pulp capping agents. This double-blinded two parallel armed randomized controlled clinical trial comprised the allocation of 108 patients with traumatically exposed dental pulp during the management of deep carious lesions by undergraduate students after fulfilling inclusion and exclusion criteria. Patients were randomized into two groups ( n  = 54 in each group) using computer-generated simple randomization, wherein one group Platelet Rich Fibrin (PRF) was prepared from patients’ blood samples and applied directly over exposed pulp followed by MTA application and in the other group MTA was applied directly over pulp exposure. In both groups, cavities were restored with resin-modified glass ionomer liner and resin composite restoration. The overall success of treatment was calculated at 6 and 12 months after assessing pulp sensibility, history of pain, tenderness on percussion and the existence of any periapical pathosis using in periapical radiographs. Moreover CBCT was used at 12 months to determine the presence or absence of dentin bridge as a secondary outcome. After 12 months follow-up, there was no statistically significant difference in overall success of pulp capping in both groups. As the both groups showed 92.59% success rate. CBCT evaluation of dentin bridge formation by Platelet Rich Fibrin (PRF) demonstrated a significantly higher percentage than that formed in cases treated with MTA alone ( p  < 0.001). Direct pulp capping with Platelet Rich Fibrin (PRF) exhibited a clinical and radiographic success rate comparable to that of MTA. Platelet Rich Fibrin (PRF) can be implemented as a direct pulp capping agent in forthcoming clinical applications.

This study aimed to determine the contribution of titanium prepared platelet-rich fibrin (T-PRF) with open flap debridement (OFD) on clinical, biochemical and radiographic measurements of periodontal regeneration. Twenty periodontitis patients with bilateral intrabony defects and stage III grade A periodontitis were included in the study. A total of 40 defects were randomly selected for OFD alone (control group, n = 20) or combined OFD+ T-PRF (test group, n = 20). Clinical and radiographic parameters (at baseline and nine months after surgery), and growth factor levels in gingival crevicular fluid (at baseline and at two, four, six, and twelve weeks after surgical treatment) were also evaluated. Considering the clinical parameters, alterations in probing pocket depth, gingival marginal level and clinical endpoint in the test regions treated with T-PRF significantly improved ( P <0.05). Fibroblast growth factor-2 and platelet-derived growth factor-BB levels between the two groups in the second and fourth weeks were also significantly different (P<0.05). Furthermore, the receptor activator of nuclear factor κB ligand/osteoprotegerin ratio between the groups was significantly different in the second, fourth, sixth, and twelfth weeks (P<0.05). The bone-filling rate was also significantly greater in the test group than in the control group (P <0.001). Compared with OFD alone, combining T-PRF with the procedure was more successful with regards to clinical, radiographic, and biochemical measurements of periodontal regeneration.

Autologous platelet-derived growth factors such as platelet rich fibrin (PRF) are receiving increasing attention in the context of different medical situations such as soft-tissue wound healing or bone regeneration in patients with cancer suffering from therapy-associated osteonecrosis. PRF has been observed to support the colonization and differentiation of osteoblasts, thereby improving the wound healing process. In the recent past, fruit extracts from the tropical plant have been associated with improved intestinal health and anti-inflammatory therapeutic effects. The aim of this study was to investigate the influence of -derived noni juice on clinical blood parameters and the composition of human PRF. Forty healthy volunteers participated in a prospective, single-blinded, placebo-controlled, cross-over study. Participants consumed either noni juice (2 ml/kg/day) or placebo for four weeks, separated by a four-week washout. Blood samples were collected, and PRF was prepared by centrifugation. Clinical blood values were analyzed, and PRF samples were examined for growth factors, structural proteins, and cytokines using ELISA and cytokine arrays. Noni juice consumption led to significant changes in blood calcium, ALAT, and γ-GT levels. PRF analysis revealed elevated interleukin-11 (IL-11), macrophage colony-stimulating factor (M-CSF), and chemokine CCL7, indicating that noni juice alters the molecular profile of PRF. Regular intake of noni juice influences PRF composition and modulates hepatic enzymes. These findings highlight the potential of dietary factors to impact regenerative biomaterials and warrant further targeted investigations.

This randomized controlled trial evaluated the efficacy of titanium-prepared platelet-rich fibrin (T-PRF) for alveolar ridge preservation (ARP) compared with leukocyte and platelet rich fibrin (L-PRF) and spontaneous healing. Thirty single-rooted teeth requiring extraction were randomly assigned to one of three groups (ARP with T-PRF, ARP with L-PRF, and spontaneous healing) using a flapless, atraumatic extraction approach. Radiographic measurements (bone width, height, and density) were obtained pre-operatively and at four months via OnDemand3D software, and clinical parameters—including post-operative pain, analgesic consumption, soft-tissue healing, periodontal probing depth, gingival recession, and keratinized tissue width—were assessed. At four months, the T-PRF group demonstrated substantially greater preservation of ridge dimensions ( P  = 0.000), and higher bone density ( P  = 0.000), along with improved soft-tissue healing ( P  < 0.05), lower pain scores ( P  = 0.000), and reduced analgesic use ( P  < 0.05). Periodontal parameters remained stable across the groups, except for keratinized tissue width (KTW), which was substantially greater in the T-PRF group ( P  = 0.020). There were substantial differences among the groups in the need for bone regeneration when implants were placed ( P  < 0.05). These findings support the clinical advantage of T-PRF in optimizing post-extraction outcomes for implant site development. This trial was registered in the UK’s Clinical Study Registry ISRCTN (ISRCTN60191547).

The present study investigated the tissue reaction of platelet rich fibrin and chitosan hydrogel as internal matrices in repairing furcal perforations in mature dogs’ teeth. Seventy-two teeth in six mongrel dogs were experimented in this study. After access opening, root canal preparation was completed and obturation was done using gutta percha/resin sealer. Furcation perforations were done, and the experimental teeth were classified according to the perforation repair protocol to three experimental groups and a positive control group (18 teeth each). Group 1: Platelet-rich fibrin matrix with premixed calcium silicate-based bioceramic putty (BC putty), Group 2: Chitosan hydrogel matrix with BC putty, Group 3: BC putty alone and Group 4: a positive control group where no repair material was utilized. Access openings were restored with composite filling. The experimented teeth and the supporting bone were sectioned into blocks and histologically examined for tissue reaction at one and three months. Statistical analysis was performed using Chi-square test, where the significance level was set at P  ≤ 0.05. BC putty and BC putty with PRF matrix exhibited less bone loss, epithelial proliferation and inflammatory reaction compared to chitosan hydrogel at one and three months intervals, also they showed more hard tissue deposition compared to chitosan hydrogel at 3-month interval. Although BC putty presented higher sealing ability with great area of newly formed hard tissue compared to chitosan hydrogel, BC putty with PRF can be considered as a successful management option for furcal perforation repair. Management of perforation is considered a challenging procedure especially when located in the furcation area, however histological evaluation of the tissue reaction to different internal matrices materials could provide favorable clinical outcomes concerning the perforation repair procedures.