Effect of platelet-rich plasma on the healing of intrabony defects treated with an anorganic bovine bone mineral: a pilot study

BACKGROUND: Periodontal therapy using the combination of platelet-rich plasma (PRP) and different grafting materials has been suggested as a modality to enhance the outcome of regenerative surgery. In most clinical studies, a barrier membrane was used to cover the defects, and thus, the effects of PRP may have been masked by the effects of the barrier. The data from controlled clinical studies evaluating the effect of regenerative therapy using various grafting materials with or without PRP are still limited. The purpose of this study was to clinically compare the healing of intrabony defects treated with either a combination of an anorganic bovine bone mineral (ABBM) and PRP to those obtained with ABBM alone. METHODS: Thirty patients with advanced chronic periodontal disease and displaying one intrabony defect were randomly treated with PRP + ABBM or ABBM alone. The following clinical parameters were evaluated at baseline and 1 year after treatment: plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), gingival recession (GR), and clinical attachment level (CAL). The primary outcome variable was CAL. RESULTS: No statistical significant differences in any of the investigated parameters between the two groups were observed at baseline. Healing was uneventful in all patients. In the PRP + ABBM group, mean PD decreased from 8.6 +/- 1.8 mm to 3.4 +/- 1.4 mm (P <0.001) and mean CAL changed from 9.9 +/- 1.7 mm to 5.3 +/- 1.8 mm (P <0.001). In the ABBM group, mean PD decreased from 8.5 +/- 2.0 mm to 3.2 +/- 1.3 mm (P <0.001) and mean CAL changed from 9.6 +/- 1.9 mm to 4.9 +/- 1.5 mm (P <0.001). CAL gains >or=3 mm were measured in 80% (12 of 15 defects) of cases treated with PRP + ABBM and in 87% (13 of 15 defects) of cases treated with ABBM alone. No statistically significant differences in any of the investigated parameters were observed between the two groups at the 1-year reevaluation. CONCLUSIONS: Within the limits of the present study, it can be concluded that 1) at 1 year after regenerative surgery with PRP + ABBM and ABBM alone, significant PD reductions and CAL gains were found, and 2) the use of PRP failed to improve the results obtained with ABBM alone.

Five-year results evaluating the effects of platelet-rich plasma on the healing of intrabony defects treated with enamel matrix derivative and natural bone mineral

BACKGROUND Regenerative periodontal surgery using the combination of enamel matrix derivative (EMD) and natural bone mineral (NBM) with and without addition of platelet-rich plasma (PRP) has been shown to result in substantial clinical improvements, but the long-term effects of this combination are unknown. METHODS The goal of this study was to evaluate the long-term (5-year) outcomes after regenerative surgery of deep intrabony defects with either EMD + NBM + PRP or EMD + NBM. Twenty-four patients were included. In each patient, one intrabony defect was randomly treated with either EMD + NBM + PRP or EMD + NBM. Clinical parameters were evaluated at baseline and 1 and 5 years after treatment. The primary outcome variable was clinical attachment level (CAL). RESULTS The sites treated with EMD + NBM + PRP demonstrated a mean CAL change from 10.5 ± 1.6 to 6.0 ± 1.7 mm (P <0.001) at 1 year and 6.2 ± 1.5 mm (P <0.001) at 5 years. EMD + NBM-treated defects showed a mean CAL change from 10.6 ± 1.7 to 6.1 ± 1.5 mm (P <0.001) at 1 year and 6.3 ± 1.4 mm (P <0.001) at 5 years. At 1 year, a CAL gain of ≥4 mm was measured in 83% (10 of 12) of the defects treated with EMD + NBM + PRP and in 100% (all 12) of the defects treated with EMD + NBM. Compared to baseline, in both groups at 5 years, a CAL gain of ≥4 mm was measured in 75% (nine of 12 in each group) of the defects. Four sites in the EMD + PRP + NBM group lost 1 mm of the CAL gained at 1 year. In the EMD + NBM group, one defect lost 2 mm and four other defects lost 1 mm of the CAL gained at 1 year. No statistically significant differences in any of the investigated parameters were observed between the two groups. CONCLUSIONS Within their limits, the present results indicate that: 1) the clinical outcomes obtained with both treatments can be maintained up to a period of 5 years; and 2) the use of PRP does not appear to improve the results obtained with EMD + NBM.

Use of platelet-rich plasma under autogenous onlay bone grafts

Objectives: To analyze the healing of autogenous onlay bone grafts in three different situations, focusing on the interface area.Material and methods: Sixteen rabbits underwent autogenous bone graft surgeries in the calvaria. The block bone grafts were positioned in three different situations: direct contact between bone graft and receptor bed, graft interposed by particulate bone, and graft interposed by platelet-rich plasma (PRP). After 7, 15, 30, and 60 days, the specimens were retrieved for histological and morphometric evaluation.Results: All groups healed uneventfully and presented incorporation of the grafts after 30 days. A slightly more evident new bone formation could be observed in the PRP group in the first analyzed period, and an earlier maturation of bone in the last period, although no statistically significant differences were achieved.Conclusion: the use of additional material between the bone graft and the receptor bed when using the onlay technique must be carefully considered, taking into account the size of the reconstruction and the cost/benefit relation. The addition of PRP in between autogenous bone blocks and the receptor bed did not confer significant benefit for the new bone formation and healing on the calvaria of bone of rabbits.

Effect of Platelet-Rich Plasma on Peri-Implant Bone Repair: A Histologic Study in Dogs

The present study evaluated the effect of platelet-rich plasma (PRP) on pen-implant bone healing. A total of 9 mongrel dogs received 36 dental implants with sandblasted acid-etched surface in lower jaws in a split-mouth design: in the PRP group (n = 18 implants) the implants were placed in association with PRP, and in the control group (n = 18 implants) the implants were placed without PRP. Biopsies were obtained and prepared for histologic and histometric analysis after 15, 30, and 55 days of healing. The biopsies retrieved at 15 days showed delicate bone trabeculae formed by immature bone with presence of numerous osteoblasts for both groups. At 30 days the trabeculae presented reversal lines and evident lamellar disposition, where some thread spaces were filled by bone and dense connective tissue. At 55 days, bone healing was not altered in the control group, and histologic aspects were variable for the group treated with PRP. There was no significant difference between the groups for bone-to-implant contact (P > .05). PRP did not enhance bone formation around sandblasted acid-etched implants.

Radiographic and histomorphometric analysis of bone healing using autogenous graft associated with platelet-rich plasma obtained by 2 different methods

The aim of this study was to conduct radiographic and histomorphometric analysis of bone healing in the calvaria of rabbits, using an autogenous graft associated with PRP obtained by 2 different methods. Thirty rabbits were divided into control and experimental groups. Lesions were produced in the calvaria and filled with autogenous graft ( control) or autogenous graft and PRP obtained by the Anitua or modified Sonnleitner methods. The animals were humanely killed 15 days after surgery and the calvarias were radiographed. The radiographs were digitized to assess the radiographic density. By histologic images of the lesion, the bone matrix was quantified. There were no significant differences in the radiographic density and the bone matrix area between the groups. The association of PRP with autogenous bone did not improve the healing process, irrespective of the method used early during healing.

Osteogenic potential of mesenchymal cells derived from canine umbilical cord matrix co-cultured with platelet-rich plasma and demineralized bone matrix

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

BMP implant associated with platelet-rich plasma in orbit fracture repair

Purpose: To evaluate a bone morphogenetic protein (BMP) implant with and without platelet-rich plasma (PRP), which is supposed to accelerate fracture consolidation in the orbit fracture treatment. Methods: Thirty-six white rabbits were subjected to orbital fracture and treated in three groups: BMP implant fracture repair (G1), BMP plus PRP implant fracture repair (G2), and fracture and spontaneous repair (G3). The animals were sacrificed at 7, 30, 90, and 180 days after surgery. A radiology evaluation was carried out on the 7th day after the fracture and at the sacrifice moments. After the animals' death, the orbital content material was removed and prepared for morphological and morphometric analysis. Results: Radiology suggested intramembranous and progressive cavitation and ossification without a reduction in implant size and with signs of calcium deposition; these events were confirmed by histological analysis, which showed a lymphomononuclear inflammatory reaction in G1 and G2, more intense 7 days after surgery and reducing after 30 days. Associating PRP with BMP did not accelerate bone induction. Conclusion: BMP implant promotes bone induction, integration at fracture site, scarce inflammatory reaction, and may be a good alternative in orbit fracture reconstruction. The addition of PRP to the BMP plate did not accelerate the resolution, and its use is not necessary. Copyright © Informa Healthcare USA, Inc.

Platelet-rich plasma stimulates cytokine expression and alkaline phosphatase activity in osteoblast-derived osteosarcoma cells

Objective: The aim of this study was to investigate the effects of PRP on SAOS-2 cells in terms of cytokine expression, cell activity and oxidative stress. Design: Cell line SAOS-2 (1 x 10(5) cells/mL) were grown in culture medium alpha-MEM with 10% FBS for 24 h and stimulated (or not) with PRP at concentrations of 3, 10 and 20%, LPS (E. coli, 10 g/mL) and IL-1 beta (1 mg/mL) for 24 h. The supernatant was collected and analyzed for the expression of cytokines in a panel array, ALP using a commercial kit and NO2- with Griess reaction method. Also, the cells were analyzed using Western blot for RANKL and slot blotting for nitrotyrosine expression. Result: There were no significant differences amongst the groups in terms of NO2-, protein nitrotyrosine content and RANKL expression. However, all stimuli increased ALP activity and in case of PRP, it was in a dose-dependent manner (p < 0.001). Also, all stimuli induced an increase in cytokines and chemokines expression, but only PRP promoted an increase of component C5, sICAM-1 and RANTES expression. Whilst IL-1 receptor antagonist (IL-1ra) expression was down-regulated by PRP, both LPS and IL-1 beta caused up-regulation of this cytokine. Conclusions: PRP can stimulate osteoblast activity and cytokine/chemokine release, as well as indicate some of the mediators that can (and cannot) be involved in this activation. (C) 2012 Elsevier Ltd. All rights reserved.

Evaluation of biomaterials with and without platelet-rich plasma: a histometric study using beagle dogs

Aim: To compare the alveolar bone repair process using biomaterial in dogs with and without the incorporation of platelet-rich plasma. Methods: Six beagles were used. Bilateral extractions of the three mandibular premolars were performed. Bio-Gen® was applied in the first alveolus, the clot was maintained in the second alveolus and Genox® was applied to the third alveolus. PRP was added to all alveoli on the left side only. The dogs were submitted to euthanasia after 30, 60 and 90 days and submitted to histological analysis for the determination of mean area of new bone formation. Tukey’s post test was used in the statistical analysis. Results: Significant increase in bone formation occurred in Bio-Gen® + PRP when compared with the other groups at 30 and 90 days. In the evaluation at 60 days, no statistically significant differences among the groups were found. Conclusions: The Bio-Gen® biomaterial led to the best bone repair and the combination of platelet-rich plasma accelerated the repair process.

Development of a cost-effective method for platelet-rich plasma (PRP) preparation for topical wound healing.

Platelet-rich plasma (PRP) is a volume of plasma fraction of autologous blood having platelet concentrations above baseline whole-blood values due to processing and concentration. PRP is used in various surgical fields to enhance soft-tissue and bone healing by delivering supra-physiological concentrations of autologous platelets at the site of tissue damage. These preparations may provide a good cellular source of various growth factors and cytokines, and modulate tissue response to injury. Common clinically available materials for blood preparations combined with a two-step centrifugation protocol at 280g each, to ensure cellular component integrity, provided platelet preparations which were concentrated 2-3 fold over total blood values. Costs were shown to be lower than those of other methods which require specific equipment and high-cost disposables, while safety and traceability can be increased. PRP can be used for the treatment of wounds of all types including burns and also of split-thickness skin graft donor sites, which are frequently used in burn management. The procedure can be standardized and is easy to adapt in clinical settings with minimal infrastructure, thus enabling large numbers of patients to benefit from a form of cellular therapy.

Bioglass Associated With Leukocyte-Poor Platelet-Rich Plasma in the Rabbit Maxillary Sinus: Histomorphometric, Densitometric, and Fractal Analysis

The objective of the present study was to evaluate the outcomes of autogenous bone graft (AB) and bioglass (BG) associated or not with leukocyte-poor platelet-rich plasma (LP-PRP) in the rabbit maxillary sinus (MS) by histomorphometric and radiographic analysis. Twenty rabbits divided into 2 groups (G1, G2) were submitted to sinus lift surgery. In G1, 10 MS were grafted with AB and 10 MS were grafted with BG. In G2, 10 MS were grafted with AB + LP-PRP and 10 MS were grafted with BG + LP-PRP. After 90 days, the animals were killed and specimens were obtained, x-rayed, and submitted to histomorphometric, radiographic bone density (RD) and fractal dimension analysis. Radiographic bone density mean values (SD), expressed as aluminum equivalent in mm, of AB, BG, AB + LP-PRP, and BG + LP-PRP groups were 1.79 (0.31), 2.04 (0.39), 1.61 (0.28), and 1.53 (0.30), respectively. Significant differences (P < 0.05) were observed between BG and AB, and BG + PRP and BG. Fractal dimension mean values were 1.48 (0.04), 1.35 (0.08), 1.44 (0.04), and 1.44 (0.06), respectively. Significant differences were observed between BG and AB, and AB + LP-PRP and BG. Mean values for the percentage of bone inside MS were 63.30 (8.60), 52.65 (10.41), 55.25 (7.01), and 51.07 (10.25), respectively. No differences were found. No correlations were observed among percentage of bone, RD and FD. Histological analysis showed that MS treated with AB presented mature and new bone formation. The other groups showed minor bone formation. Within the limitations of this study, the results indicated that at a 90-day time end point, AB yielded better results than AB + LP-PRP, BG, and BG + LP-PRP and should be considered the primary material for MS augmentation.

Platelet-Rich Plasma And Chronic Wounds: Remaining Fibronectin May Influence Matrix Remodeling And Regeneration Success

Background: Platelet-rich plasma has been largely used as a therapeutic option for the treatment of chronic wounds of different etiologies. The enhanced regeneration observed after the use of platelet-rich plasma has been systematically attributed to the growth factors that are present inside platelets' granules. Aim: We hypothesize that the remaining plasma and platelet-bound fibronectin may act as a further bioactive protein in platelet-rich plasma preparations. Methods: Recent reports were analyzed and presented as direct evidences of this hypotheses. Results: Fibronectin may directly influence the extracellular matrix remodeling during wound repair. This effect is probably through matrix metalloproteinase expression, thus exerting an extra effect on chronic wound regeneration. Conclusions: Physicians should be well aware of the possible fibronectin-induced effects in their future endeavors with PRP in chronic wound treatment. © 2013 International Society for Cellular Therapy.

Platelet-rich plasma (PRP): methodological aspects and clinical applications

The clinical use of platelet-rich plasma (PRP) is based on the increase in the concentration of growth factors and in the secretion of proteins which are able to maximize the healing process at the cellular level. Since PRP is an autologous biologic material, it involves a minimum risk of immune reactions and transmission of infectious and contagious diseases, and it has been widely used for the recovery of musculoskeletal lesions. Despite the great potential for applicability, the implementation of the therapeutic employment of PRP as a clinical alternative has become difficult, due to the lack of studies related to the standardization of the techniques and/or insufficient description of the adopted procedures. Therefore, it is required establish standard criteria to be followed for obtaining a PRP of high quality, as well as a larger number of studies which should establish the proper concentration of platelets for the different clinical conditions. In this context, the purpose of this review is to discuss some methodological aspects used for achieving the PRP, as well as to discuss the bioactive properties of PRP, and to point out its therapeutic use in different fields of regenerative medicine.

Healing and expression of growth factors (TGF-β and PDGF) in canine radial ostectomy gap containing platelet-rich plasma

Objectives: To evaluate bone healing in an experimental radial ostectomy in dogs treated with autologous platelet-rich plasma (PRP), through histological, densitometric, radiographic studies, as well as expression of growth factors in the ostectomy gap. Methods: Twenty-one dogs were randomly divided into either a control or a PRP group. All underwent unilateral ostectomy of the radius to generate a gap of 2.0 mm, that was stabilized with external skeletal fixation. The ostectomy gap was either filled with PRP or left empty as a control. The radiographic and densitometric studies were performed after surgery, then at intervals until 60 days during the post-surgery period. Histological and immunohistochemical evaluations were performed at seven or 60 days post-surgery. Analyses were performed using a statistical analysis system, and the level of significance was set at p <0.05. Results: The median radiographic healing score in the PRP group increased significantly between day 0 and day 60. Furthermore, at 60 days, the median healing score and the proportion of healed ostectomies in the control group (score 1; 1/6 healed) and the PRP group (score 5; 4/5 healed) were significantly different. There were differences between groups in radiographic and densitometric values at days 45 and 60. The histological evaluation showed advanced bone healing at 60 days in the PRP group and signs of delayed union in the control group. Clinical relevance: Platelet-rich plasma can be used as an adjuvant therapy because it may promote better bone healing of a radial ostectomy treated with external skeletal fixation in dogs.

Impact of platelet rich plasma and adipose stem cells on lymphangiogenesis in a murine tail lymphedema model.

BACKGROUND: Lymphedema is an underdiagnosed pathology which in industrialized countries mainly affects cancer patients that underwent lymph node dissection and/or radiation. Currently no effective therapy is available so that patients' life quality is compromised by swellings of the concerned body region. This unfortunate condition is associated with body imbalance and subsequent osteochondral deformations and impaired function as well as with an increased risk of potentially life threatening soft tissue infections. METHODS: The effects of PRP and ASC on angiogenesis (anti-CD31 staining), microcirculation (Laser Doppler Imaging), lymphangiogenesis (anti-LYVE1 staining), microvascular architecture (corrosion casting) and wound healing (digital planimetry) are studied in a murine tail lymphedema model. RESULTS: Wounds treated by PRP and ASC healed faster and showed a significantly increased epithelialization mainly from the proximal wound margin. The application of PRP induced a significantly increased lymphangiogenesis while the application of ASC did not induce any significant change in this regard. CONCLUSIONS: PRP and ASC affect lymphangiogenesis and lymphedema development and might represent a promising approach to improve regeneration of lymphatic vessels, restore disrupted lymphatic circulation and treat or prevent lymphedema alone or in combination with currently available lymphedema therapies.