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Aim: 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. Methods: 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). Results: 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. Conclusion: 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.

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BACKGROUND: Platelet concentrates like leucocyte- and platelet-rich fibrin (L-PRF) have been widely evaluated in different oral surgical procedures to promote the healing process. However, liquid L-PRF products such as liquid fibrinogen have been poorly explored, especially in the biomimetic functionalization of dental implants. The aim of this in vitro study is to evaluate the interaction between 5 different dental implant surfaces and liquid fibrinogen. METHODS: Five commercially available dental implants with different surfaces (Osseospeed™, TiUnite™, SLActive®, Ossean®, and Plenum®) were immersed for 60 minutes in liquid fibrinogen obtained from healthy donors. After this period, the implants were removed and fixed for scanning electron microscopy (SEM). RESULTS: All dental implants were covered by a fibrin mesh. However, noticeable noncontact areas were observed for the Osseospeed™, TiUnite™, and SLActive® surfaces. On the other hand, Ossean® and Plenum® surfaces showed a dense and uniform layer of fibrin covering almost the entire implant surface. The Osseospeed™, TiUnite™, and SLActive® surfaces presented with lower blood cell numbers inside the fibrin mesh compared with the others. Moreover, at higher magnification, thicker fibrin fibers were observed in contact with Ossean® and Plenum® surfaces. The Plenum ®surface showed the thickest fibers which also inserted and interconnect to the microroughness. CONCLUSION: The initial contact between an implant surface and the fibrin network differs significantly among different implant brands. Further studies are necessary to explore the clinical impact of these observations in the osseointegration process of dental implants.

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OBJECTIVES: The aim of this study was to describe the histological and clinical outcome of "dentin block" (a mixture of autologous particulate dentin, leukocyte- and platelet-rich fibrin (L-PRF), and liquid fibrinogen) in alveolar ridge preservation. MATERIAL AND METHODS: Ten extraction sockets were grafted with "dentin block," a mixture of particulate autologous dentin with chopped leukocyte-platelet-rich fibrin (L-PRF) membranes at a 1:1 ratio, and liquid fibrinogen as a binder. Two grafted sites were followed at 4 and 5 months, and 6 sites at 6 months. Biopsies were taken from the core of the grafted site for histologic and histo-morphometric analysis. RESULTS: All patients completed the study without any adverse event. The vertical and horizontal dimensions of the alveolar ridge were preserved or even increased after 4, 5, or 6 months and remained stable after 6 months of the implant placement. The histological examination revealed a median relative percentage of bone, dentin, and connective tissue of 57.0, 0.9, and 39.3%, respectively. A comparison of samples at different time points (4, 5, and 6 months) showed a progressive increase in the proportion of bone with a decrease in the proportion of dentin. The bone was compact with normal osteocytes and moderate osteoblastic activity. In 4 out of 10 samples, no dentin was observed; in the other samples, it represented 1-5% (with geometric fragments). CONCLUSIONS: Dentin block showed to be a suitable bone substitute in an alveolar ridges preservation model. CLINICAL RELEVANCE: The promising results of dentin block as a bone substitute in alveolar ridge preservation could have an important clinical impact considering this biomaterial brings together the regenerative potential of three autologous products with excellent biological and clinical behavior, low risk of adverse effects, and feasible acquisition.

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AIM: To evaluate the impact of two non-surgical periodontal treatment modalities on metabolic and periodontal clinical parameters in subjects with type 2 diabetes mellitus (T2DM) and poor glycaemic control and chronic periodontitis. MATERIAL AND METHODS: A randomized controlled clinical trial was conducted. Ninety-three T2DM subjects with glycosylated haemoglobin (HbA1c) > 7% were randomly assigned to one of two groups receiving scaling with root planing in multiple sessions quadrant-by-quadrant (Q by Q) or within 24 hr (one stage). Periodontal parameters, HbA1c, glycaemia blood levels (FPG) and C-reactive protein (CRP) values were assessed at baseline and at 3 and 6 months post-therapy. RESULTS: At 6 months, HbA1c had decreased by 0.48% in the Q by Q group and by 0.18% in the one-stage group (p = 0.455). After therapy, subjects with an initial HbA1c < 9% showed an increase of 0.31% (p = 0.145), compared with a decrease of 0.88% (p = 0.006) in those with an initial HbA1c ≥ 9%. Periodontal parameters improved significantly (p < 0.0001) post-therapy, with similar results for both treatment modalities. CONCLUSION: Periodontal therapy had the greatest impact on HbA1c reduction on patients with an HbA1c > 9% regardless of treatment modality. Both modalities resulted in significant improvements in periodontal parameters.

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Chronic wounds (VLU: venous leg ulcer, DFU: diabetic foot ulcer, PU: pressure ulcer, or complex wounds) affect a significant proportion of the population. Despite appropriate standard wound care, such ulcers unfortunately may remain open for months or even years. The use of leukocyte- and platelet-rich fibrin (L-PRF) to cure skin ulcers is a simple and inexpensive method, widely used in some countries but unknown or neglected in most others. This auto-controlled prospective cohort study explored and quantified accurately for the first time the adjunctive benefits of topical applications of L-PRF in the management of such refractory ulcers in a diverse group of patients. Forty-four consecutive patients with VLUs (n = 28, 32 wounds: 17 ≤ 10 cm2 and 15 > 10 cm2), DPUs (n = 9, 10 wounds), PUs (n = 5), or complex wounds (n = 2), all refractory to standard treatment for ≥3 months, received a weekly application of L-PRF membranes. L-PRF was prepared following the original L-PRF method developed more than 15 years ago (400g, 12 minutes) using the Intra-Spin L-PRF centrifuge/system and the XPression box kit (Intra-Lock, Boca Raton, FL, USA; the only CE/FDA cleared system for the preparation of L-PRF). Changes in wound area were recorded longitudinally via digital planimetry. Adverse events and pain levels were also registered. All wounds showed significant improvements after the L-PRF therapy. All VLUs ≤ 10 cm2, all DFUs, as well as the two complex wounds showed full closure within a 3-month period. All wounds of patients with VLUs > 10 cm2 who continued therapy (10 wounds) could be closed, whereas in the five patients who discontinued therapy improvement of wound size was observed. Two out of the five PUs were closed, with improvement in the remaining three patients who again interrupted therapy (surface evolution from 7.35 ± 4.31 cm2 to 5.78 ± 3.81 cm2). No adverse events were observed. A topical application of L-PRF on chronic ulcers, recalcitrant to standard wound care, promotes healing and wound closure in all patients following the treatment. This new therapy is simple, safe and inexpensive, and should be considered a relevant therapeutic option for all refractory skin ulcers.

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More insight into the biological fundamentals of leukocyte platelet-rich fibrin (L-PRF) guided healing is necessary to recommend its application, in particular in deficient bone sites that need to support implants. This study investigated the short-term bone healing effect of L-PRF treatment in cylindrical non-critical sized bone defects with 3 mm diameter and 6 mm depth in tibiae of 18 adult male New Zealand White rabbits. After a randomization process, 96 bone defects were prepared and half of them were filled with a L-PRF membrane, while untreated defects in the opposite tibia served as control group. The rabbits were euthanized after 7, 14 or 28 days of healing. The bone healing of the cortical and medullary areas was investigated by micro-CT, while the expression of molecular markers (RUNX2, VEGFA, COL1A2 and BMP2) was assessed by qRT-PCR. Treatment with L-PRF did not affect the micro-structural bone characteristics of the repaired bone tissue, except for a decrease in the trabecular connectivity at the cortical level after 14 days of healing. At this time, RUNX2 and VEGFA mRNA levels were significantly lower in the treated defects. L-PRF membranes thus had a temporary negative influence on the bone microarchitecture (Tb.Pf) and on the RUNX2 and VEGFA expression during early bone healing. Overall, L-PRF treatment did not enhance bone regeneration in these non-critical size defects after 28 days.

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OBJECTIVES: To evaluate clinical, radiographic, microbiologic, and biomechanical parameters related to bone remodeling around implants with external hexagon (EH) and Morse-taper (MT) connections. MATERIALS AND METHODS: Twelve totally edentulous patients received four custom-made implants in the interforaminal region of the mandible. Two of those implants had the same macroscopic design, but different prosthetic connections. All patients received an immediate implant-supported prosthesis. Clinical parameters (periimplant probing pocket depth (PPD), modified gingival index (mGI), and mucosal thickness (MTh)) were evaluated at 12 months follow-up. The distance between the top of the implant and the first bone-to-implant contact (IT-FBIC) was evaluated on standardized digital peri-apical radiographs acquired at 1, 3, 6, and 12 months follow-up. Samples of the subgingival microbiota were collected 1, 3, and 6 months after implant loading and used for the quantification of Tanerella forsythia, Porphyromonas gingivalis, Aggragatibacter actinomycetemcomitans, Prevotella intermedia, and Fusobacterium nucleatum. Further, 36 computerized-tomography based finite element (FE) models were accomplished, simulating each patient under three loading conditions. RESULTS: The evaluated clinical parameters were equal for EH and MT implants. Mean IT-FBIC was significantly different between the tested connections (1.17 ± 0.44 mm for EH, and 0.17 ± 0.54 mm for MT, considering all evaluated time periods). No significant microbiological differences could be observed between tested connections. FE analysis showed a significantly higher peak of equivalent (EQV) strain (p = 0.005) for EH (mean 3,438.65 µÎµ) compared to MT (mean 840.98 µÎµ) connection. CONCLUSIONS: Radiographic periimplant bone loss depends on the implant connection type. MT connections showed less periimplant bone loss, compared to EH connections.

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OBJECTIVES: This clinical trial explored the clinical and radiographic dimensional changes of the alveolar ridge in the first 4 months after tooth extraction in combination with the application of Leukocyte- and Platelet Rich Fibrin (L-PRF). METHODS: Eighteen single rooted maxillary and mandibular sockets were filled with L-PRF without soft tissue closure. Clinical measurements (bone sounding) were performed using a customized acrylic stent and radiographic measurements were accomplished using Cone Beam Computed Tomography (CBCT), immediately after tooth extraction and after 4 months. RESULTS: The clinical observations indicated a mean horizontal resorption of 1.18±2.4mm (p=0.8) at the crest, 1.25±2.0mm (p=0.57) and 0.83±2.0mm (p=0.78) at 2mm and 4mm apical to the crest, respectively. The buccal plate demonstrated a mean vertical loss of 0.44±3.5mm (p=0.9), the centre of the socket had a significant filling of 5.72±3.6mm (p=0.0001) and the oral cortical plate had a mean vertical gain of 0.09mm±1.57mm (p=0.9). The radiographic analysis demonstrated a mean vertical bone loss of 0.27±2.5mm (p=0.9) on the buccal and of 0.03±1.6mm (p=0.9) at the oral crest. The width of the alveolar ridge had a mean loss of 1.33mm±1.43mm. CONCLUSIONS: Within the limitations of this pilot study, it can be concluded that L-PRF might show clinical benefits for ridge preservation.

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BACKGROUND: A broader understanding of the immune inflammatory profile of peri-implant diseases could be helpful in the development of host-targeted preventive and therapeutic strategies. The aim of this study is to answer two clinical questions: 1) whether patients with peri-implantitis (PP) present higher prevalence of any specific inflammatory cytokine in peri-implant crevicular fluid (PICF) compared with healthy patients; and 2) whether local inflammation measured in PICF can be used as a predictor for incipient PP. METHODS: A systematic review of the literature on the most common cytokines released in PICF in healthy and PP-affected sites was conducted from 1996 up to and including October 2013 using predefined search strategies. Cross-sectional and prospective longitudinal studies were considered. Meta-analyses were done separately for healthy, mucositis (MU), and PP outcomes. RESULTS: Interleukin (IL)-1ß was the most studied cytokine (n = 12), followed by tumor necrosis factor (TNF)-α (n = 10). Other cytokines were also linked to PP, such as IL-4, IL-6, IL-8, IL-10, IL-12, and IL-17. Statistical differences were revealed when IL-1ß release was compared between healthy implant sites and PP (P = 0.001) or MU sites (P = 0.002), respectively; when PP and MU were compared, no statistical differences could be detected (P = 0.80). For TNF-α release, significant differences were found between healthy and PP implants (P = 0.02). CONCLUSIONS: PICF containing inflammatory mediators, such as IL-1ß and TNF-α, can be used as additional criteria for a more robust diagnosis of peri-implant infection. Additionally, once the inflammatory process is installed, no differences were found between peri-implant MU and PP.

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