Platelet-rich plasma (PRP) is a popular treatment for equine tendon and ligament injuries; however, commercial PRP systems are expensive. Development of a safe, inexpensive alternative would make PRP therapy more widely available to horse owners. The purpose of this study was to evaluate the quality and bacteriologic safety of PRP produced by three simple, inexpensive tube centrifugation methods and compare the results to a commercial system. Citrated blood collected from 26 normal horses was processed by four methods: blood collection tubes centrifuged at 1200 and 2000 x g, a 50ml conical tube, and a commercial system. Platelet and cell counts and mean platelet volume (MPV) in whole blood and PRP were determined using an automated hematology analyzer. Results were analyzed using mixed model ANOVA with post-hoc comparisons (MPV and fold change for RBC, WBC, and platelets) and binary logistic generalized estimating equations with horse as a blocking factor (absolute numbers of WBC, and platelets). Aerobic and anaerobic cultures were performed. Significance was set at p<0.05. Mean platelet concentrations ranged from 1.55 to 2.58 fold. The conical tube method produced the highest number of PRP samples with platelet concentrations of greater than 2.5-fold and within the clinically acceptable range of >250,000 platelets/?l. WBC counts were lowest using the commercial system and unacceptably high using the red top methods. The incidence of bacterial contamination was low (2.1%). Based on these results, the conical tube method may be a suitable alternative to commercial PRP systems in cases with budgetary constraints. / Master of Science

Thesis (MSD) --Boston University, Henry M. Goldman School of Dental Medicine, 2009 (Department of Periodontology and Oral Biology). / Includes bibliographic references: leaves 52-59. / Platelet-rich plasma (PRP) is a concentrated gel of platelets that contains several growth factors. Growth factors have been recognized as the part of PRP that play role in regeneration of the bone. It is not clear how these growth factors in PRP affect the bone regeneration. Resolvin El (RvEl; 5S,12R,18R-trihydroxyeicosapentaenoic acid) is an pro-resolving lipid mediator derived from omega-3 fatty acid eicosapentaenoic acid and shown to have potent effects on the resolution of inflammation. The purpose of this study was to analyze the action of PRP and RVEl on the proliferation and behavior of osteoblasts and osteoclasts in vitro. PRP was prepared from 14 healthy donors. Osteoblast cultures were from a cell line (Saos2) of osteosarcoma cells. Osteoclasts were differentiated from primary human peripheral blood monocytes. Osteoclastic morphology was studied and activity was analyzed via resorption on dentin discs using SEM. PRP and RVE 1 were added at different doses and time-points. Osteoblast function was analyzed by osteocalcin expression and release. Osteoclast activity was assessed by resorption and cathepsin K expression. PRP and RvEl comparably increased the osteoblastic activity and suppressed the osteoclast differentiation and function. These results suggest that multiple tools are available to reverse the inflammation and restore the lost bone architecture as a result of periodontal disease.

Resumo: Lesões na cartilagem articular representam um desafio para a Medicina Veterinária devido ao limitado potencial intrínseco de reparação. As células tronco mesenquimais (CTM) e o plasma rico em plaquetas (PRP) vêm sendo utilizados na reparação condral pela capacidade de imunomodulação, quimiotaxia, por diminuir a degradação da cartilagem articular e exercer efeito anti-inflamatório. Esse trabalho estudou a eficácia das CTM e do PRP no tratamento de lesões condrais articulares, experimentalmente induzidas em equinos, apresentando os benefícios e os principais obstáculos encontrados na aplicação clínica e cirúrgica de equinos. Para isso, foi induzida uma lesão condral, na tróclea medial femoral dos dois membros pélvicos de oito animais. Após 30 dias da indução dessa lesão os animais, divididos em quatro grupos, receberam os tratamentos propostos: com CTM (Grupo 1); PRP (Grupo 2); CTM e PRP (Grupo 3) e um controle sem tratamento (Grupo 4). As avaliações dos parâmetros clínico-laboratoriais foram realizadas antes da indução da lesão (T-0), quinzenalmente até 120 dias e aos 150 dias (T-150). Avaliações macroscópicas, histopatológicas e histoquímicas foram realizadas em T-0 e T- 150. Pôde-se observar discreta melhora clínico-laboratorial dos grupos tratados quando comparados ao grupo controle. Os exames macroscópicos, histopatológicos e histoquímicos revelaram um melhor tecido de reparação nos grupos tratados, principalmente nos grupos 2 e 3. Sendo assim, o implante autólogo de CTM e do PRP apresentou efeitos benéficos no tratamento de lesões condrais, experimentalmente induzidas em equinos / Abstract: Articular cartilage defects represent a challenge for Veterinary Medicine due to the limited intrinsic potential of repair. Mesenchymal Stem Cells (MSC) and Platelet-Rich Plasma (PRP) have been used in cartilage repair to promote immunomodulation, chemotaxis, to decrease the degradation of articular cartilage and to take antiinflammatory effect. This work aims to study the use of intralesional MSC and the PRP in the treatment of articular cartilage defects induced experimentally in horses, noting the benefits and challenges encountered in implementing clinical-surgical in horses. For this purpose, both patellofemoral joints of eight animals were approached by arthroscopic surgery to perform a cartilage defect, on the medial femoral trochlea. After 30 days of induction the animals, who were divided into four groups, received treatment with: MSC (Group 1), PRP (Group 2), MSC and PRP (Group 3) and a control without treatment (Group 4). The clinical and laboratory assessments were performed every 15 days in 120 days, with last analysis in day 150. The macroscopic and morphologic analyses were performed at 0 and 150 day. There were clinical and laboratory improvement in the treated groups compared with control group. The macroscopic and morphological analysis showed a better tissue repair in the treated groups, mainly in groups 2 and 3. Thus, the implantation of autologous MSC and PRP showed beneficial effects in the treatment of chondral lesions, experimentally induced in horses / Orientador: Ana Liz Garcia Alves / Coorientador: Marcos Jun Watanabe / Banca: Ligia Souza Lima Silveira da Mota / Banca: Raquel Yvone Arantes Baccarin / Mestre

Made available in DSpace on 2014-06-11T19:23:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-06-14Bitstream added on 2014-06-13T18:48:31Z : No. of bitstreams: 1 yamada_alm_me_botfmvz.pdf: 8714662 bytes, checksum: 3c5312366f54f7619e28b40986950ace (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Lesões na cartilagem articular representam um desafio para a Medicina Veterinária devido ao limitado potencial intrínseco de reparação. As células tronco mesenquimais (CTM) e o plasma rico em plaquetas (PRP) vêm sendo utilizados na reparação condral pela capacidade de imunomodulação, quimiotaxia, por diminuir a degradação da cartilagem articular e exercer efeito anti-inflamatório. Esse trabalho estudou a eficácia das CTM e do PRP no tratamento de lesões condrais articulares, experimentalmente induzidas em equinos, apresentando os benefícios e os principais obstáculos encontrados na aplicação clínica e cirúrgica de equinos. Para isso, foi induzida uma lesão condral, na tróclea medial femoral dos dois membros pélvicos de oito animais. Após 30 dias da indução dessa lesão os animais, divididos em quatro grupos, receberam os tratamentos propostos: com CTM (Grupo 1); PRP (Grupo 2); CTM e PRP (Grupo 3) e um controle sem tratamento (Grupo 4). As avaliações dos parâmetros clínico-laboratoriais foram realizadas antes da indução da lesão (T-0), quinzenalmente até 120 dias e aos 150 dias (T-150). Avaliações macroscópicas, histopatológicas e histoquímicas foram realizadas em T-0 e T- 150. Pôde-se observar discreta melhora clínico-laboratorial dos grupos tratados quando comparados ao grupo controle. Os exames macroscópicos, histopatológicos e histoquímicos revelaram um melhor tecido de reparação nos grupos tratados, principalmente nos grupos 2 e 3. Sendo assim, o implante autólogo de CTM e do PRP apresentou efeitos benéficos no tratamento de lesões condrais, experimentalmente induzidas em equinos / Articular cartilage defects represent a challenge for Veterinary Medicine due to the limited intrinsic potential of repair. Mesenchymal Stem Cells (MSC) and Platelet-Rich Plasma (PRP) have been used in cartilage repair to promote immunomodulation, chemotaxis, to decrease the degradation of articular cartilage and to take antiinflammatory effect. This work aims to study the use of intralesional MSC and the PRP in the treatment of articular cartilage defects induced experimentally in horses, noting the benefits and challenges encountered in implementing clinical-surgical in horses. For this purpose, both patellofemoral joints of eight animals were approached by arthroscopic surgery to perform a cartilage defect, on the medial femoral trochlea. After 30 days of induction the animals, who were divided into four groups, received treatment with: MSC (Group 1), PRP (Group 2), MSC and PRP (Group 3) and a control without treatment (Group 4). The clinical and laboratory assessments were performed every 15 days in 120 days, with last analysis in day 150. The macroscopic and morphologic analyses were performed at 0 and 150 day. There were clinical and laboratory improvement in the treated groups compared with control group. The macroscopic and morphological analysis showed a better tissue repair in the treated groups, mainly in groups 2 and 3. Thus, the implantation of autologous MSC and PRP showed beneficial effects in the treatment of chondral lesions, experimentally induced in horses

Platelet-rich plasma (PRP) is the cell free plasma, which has an enriched concentration of platelets and clotting factors with the ability to enhance the natural healing process. PRP is often used by physicians in an office setting to accelerate the healing of a variety of sports related injuries, chronic wounds and enhance skin rejuvenation. PRP mimics the wound healing cascade by enhancing the recruitment, proliferation and differentiation of cells involved in tissue regeneration. Although PRP is used to enhance healing, the efficacy thereof is debated as no clear-cut set of parameters is available that device manufacturers and protocols should follow. The lack of uniformity in the PRP preparation methods results in differing PRP volume, platelet contents and unavoidably platelet-derived growth factors. Therefore, the aim of this study was to develop a simple and rapid method for preparing autologous PRP in an office setting using a tabletop centrifuge for point-of-care use. The simplified preparation procedure involved a single centrifugation step of 18 ml of whole blood, which sufficiently enriched the platelet content in the PRP fraction. As activated platelets express and release growth factors and cytokines that mediate the different phases of the wound healing cascade, the extracted PRP fraction was activated with an ethanol, calcium chloride (CaCl2) and platelet poor plasma (PPP) preparation in glass containers, without the collection of additional blood as required in some protocols. The activated PRP formed a fibrin clot, trapping the degranulating platelets and its released growth factors. The concentration of TGF- 1 obtained from the fibrin clot was 45.49 ± 3.80 ng/ml, in range with the available literature. During the in vitro studies, the extracted PRP by the developed method was able to significantly induce cell proliferation in a dose dependent manner. Cells enumerated with the crystal violet assay indicated that the cells treaded with 5% or 10% PRP significantly increased the percentage of viable cells to 165-176% and 156-158%, when compared to the positive controls. Cells enumerated with the MTT-assay indicated that the cells treaded with 5% or 10% PRP increased the percentage of viable cells to 79-91% and 87-105% which is comparable to that of the positive control. Data from the cellular proliferation assays indicate that sufficient plateletderived growth factors had been obtained with the preparation procedure. Furthermore, data from the in vivo studies indicated that the extracted PRP was able to augment soft tissue regeneration and bone formation. Treatment with the activated PRP resulted in symptom reduction and accelerated healing of various injuries. The simplified preparation and the use of the provided study product packaged in a kit developed during this study will enable physicians to easily obtain autologous PRP, in an office setting for point-of-care use, with the ability to induce tissue regeneration. / Dissertation (MSc)--University of Pretoria, 2013 / gm2014 / Pharmacology / unrestricted

Platelet rich plasma (PRP) is a popular autologous biological therapy used for the treatment of various equine ailments, including tendon and ligament injuries, osteoarthritis, and cutaneous wounds. A number of commercial products are available for producing PRP, each generating a slightly different product. Variations in platelet numbers and white blood cell (WBC) counts are believed to be the most critical variables, as they are directly related to concentrations of growth factors and inflammatory cytokines. Accurate documentation of platelet numbers is essential for prospective evaluation of clinical outcomes, but can be problematic in platelet concentrates depending on the counting method employed. The objectives of this study were to compare the performance of four platelet counting technologies in equine platelet concentrates and to evaluate the ability of the Magellan PRP system to concentrate equine platelets. We hypothesized that there would be no differences in platelet counts among the four counting technologies and that the Magellan system would generate platelet concentrations greater than 500,000/μL. Citrated whole blood was collected from 32 horses and platelet, WBC, and red blood cell concentrations were measured using a commercial hematology analyzer (Advia 2120) prior to preparation of PRP using the Magellan system. Platelets were quantified in individual identical aliquots of equine PRP produced by the Magellan system (n=32) using three different technologies: optical scatter (Advia 2120), impedance (CellDyn 3700), and hand count using direct microscopy (Thrombo-TIC). An immunofluorescent counting method was performed on a subset of 15 of the 32 samples using a mouse monoclonal anti-sheep antibody against integrin alpha αIIbβ₃ (anti-CD41/CD61) and a fluorescent secondary antibody. Measured platelet concentrations were compared using Passing and Bablok regression analyses and mixed model ANOVA. The Magellan PRP system yielded mean (± SD) platelet and WBC counts of 893,090 ± 226,610/μL and 35,806 ± 9,971/μL, respectively. Platelet counts generated by optical scatter were consistently higher than those generated by impedance. Systematic and proportional biases were observed between these two automated methods. No bias (systematic or proportional) was observed among any of the other counting methods. Despite the bias detected between the two automated systems, there were no significant differences on average among the four counting methods evaluated, based on the ANOVA. All four platelet counting methods tested are therefore suitable for quantifying platelets in equine PRP for clinical applications. The Magellan PRP system consistently generated desirably high platelet concentrations as well as higher than expected WBC concentrations. The high platelet concentrations served as a good test medium for the study; however, the concurrent high WBC counts may be undesirable for selected orthopedic applications. / Master of Science

The field of tissue engineering aims to develop viable substitutes with the ability to repair and regenerate the functions of damaged tissue. Common practices to supplement bone regeneration in larger defects include bone graft biomaterials such as autografts, allografts, xenografts, and synthetic biomaterials. Autologous bone grafting is the current gold-standard procedure used to replace missing or damaged bone. However, these grafts have disadvantages such as donor site morbidity, limited availability, and the need for a secondary surgery. The focus of this study is to tissue engineer a lyophilized gelatin composite sponge composed of hydroxyapatite (HA), chitin whiskers (CW), and preparations rich in growth factors (PRGF) to provide sufficient structural support to the defect site while enhancing the body’s own reparative capacity, ultimately eliminating the need for autologous tissue harvesting or repeat operations. The present study investigates several in vitro evaluations on multiple compositions of modified gelatin sponge scaffolds for use in bone graft applications. Gelatin sponges were fabricated via freeze-drying, enhanced with PRGF, HA, and/or CW, and cross-linked with 50 mM 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) either during or post gelation. Initial evaluation of all scaffold combinations indicated that scaffolds released contents up to 90 days, EDC cross-linking during gelation allowed for more protein release, and had the ability to swell. Since the incorporation of PRGF, HA, and CW increased cell infiltration, and production of cell-created mineral matrix over 90 days in culture, these scaffolds were further characterized. Freeze-dried gelatin sponges enhanced with PRGF, HA, and CW and cross-linked during gelation with EDC (PHCE) were mineralized (M) in 5x revised simulated body fluid (r-SBF) for 1 hour to create a bone-like mineral surface. Gelatin EDC scaffold controls (GE), GE-M, PHCE, and PHCE-M scaffolds were characterized for their ability to swell, mineralizing potential, surface morphology, growth factor incorporation and release, uniaxial compression properties, and cell attachment, proliferation, infiltration, and protein/cytokine secretion.. After mineralization, scanning electron microscopy showed sparse clusters of mineral deposition for GE-M scaffolds while PHCE-M scaffolds exhibited a more uniform mineral deposition. Both GE and PHCE scaffolds were porous structures that swelled up to 50% of their original volume upon hydration. Over 21 days incubation, PHCE-M scaffolds cumulatively released about 30% of their original protein content, significantly more than all other scaffolds. Multiplex Luminex assays confirmed the successful incorporation of PRGF growth factors within PRGF sponges. For acellular uniaxial compression testing, PHCE-M scaffolds reported lower Young’s modulus values (1.3 - 1.6 MPa) when compared to GE and GE-M scaffolds (1.6 – 3.2 MPa). These low modulus values were comparable to values of tissue found in early stages of bone healing. DAPI (4',6-diamidino-2-phenylindole) staining and imaging showed an increase in initial cell attachment and infiltration of PHCE and PHCE-M scaffolds on day 1. GE-M scaffolds also appeared to attach more cells than the GE control. MTS cell proliferation assay results indicated that on days 4 and 7, PHCE scaffolds increased cell proliferation (compared to GE controls). MTS also illustrated that the addition of a mineral coating increases and decreases cell proliferation on GE-M and PHCE-M scaffolds, respectively. Multiplexer analysis of MG-63 protein/cytokine secretion suggests that cells are responding in a bone regenerative fashion on all scaffolds, as evidence of osteocalcin secretion. Little to no secretion of osteopontin, IL-1β, and TNF-α demonstrates that scaffolds are not influencing cells to secrete factors associated with bone resorption. The compressive mechanical properties of cellularized scaffolds did not differ much from acellular scaffolds. The collective results indicated increased cellular attachment, infiltration, and bone regenerative protein/cytokine secretion by cells on GE-M scaffolds, which support the addition of a bone-like mineral surface on GE scaffolds. Cellularized PHCE and PHCE-M scaffolds report similar advantages as well as Young’s modulus values in the range of native tissues present in the early stages of bone healing. The results of this study propose that the developed PHCE and PHCE-M scaffolds exhibit good cellular responses and mechanical properties for use in early bone healing applications.

Os hemoderivados têm sido utilizados com frequência cada vez maior na medicina equina, sendo caracterizados como um produto autólogo, com maior quantidade de fatores de crescimento e que melhora a capacidade de cicatrização de tecidos com pouco aporte sanguíneo, como tendões e articulações, diminuindo o tempo de recuperação do animal. Sabe-se que os fatores de crescimento são derivados das plaquetas, porém a correlação positiva entre o aumento na contagem plaquetária e a maior concentração de fatores de crescimento ainda é motivo de discussão entre os autores. Com o intuito de se obter um produto final com maior contagem plaquetária, é frequente o aumento da velocidade ou número de centrifugações na metodologia empregada, aumentando o risco de agregação plaquetária precoce. O objetivo do presente trabalho é estudar o efeito da dupla centrifugação no preparo de PRP, por meio da comparação entre contagens plaquetárias, concentrações de fator de crescimento TGF-&#946;1, e grau de ativação plaquetária por meio da porcentagem de agregação. Foram utilizados 12 equinos, machos, de 3 a 5 anos, clinicamente sadios. Para tanto foram realizados dois protocolos distintos: um com centrifugação única e o outro com dupla centrifugação. No primeiro, o sangue com anticoagulante foi centrifugado a 141G/12 minutos; enquanto no segundo a primeira centrifugação foi de 300G/5 minutos seguida de 700G/15 minutos, com repouso entre as mesmas e após. Os produtos obtidos após cada centrifugação foram submetidos à contagem plaquetária, teste de agregação e quantificação de TGF--&#946;1 por meio de kit ELISA. Os resultados obtidos demonstraram maior concentração plaquetária quando utilizado protocolo de dupla centrifugação. Agregometria evidenciou maior ativação das plaquetas durante o preparo do PRP quando submetidas a maiores velocidades de centrifugação (força gravitacional) e não ao fato das amostras serem centrifugadas duas vezes. A quantificação do TGF--&#946;1 não mostrou diferença quando realizado em amostras com apenas uma centrifugação, mas demonstrou valores maiores no produto final da segunda centrifugação. A avaliação por meio de coeficiente de determinação e coeficiente de correlação de Pearson evidenciou correlação positiva entre contagem plaquetária e de TGF--&#946;1. O protocolo com dupla centrifugação se mostrou mais eficaz em concentrar plaquetas e TGF--&#946;1, não sendo prejudicado pela ativação precoce dessas plaquetas durante o preparo. / Blood derived products have been used in equine medicine with increasing frequency, being characterized as an autologous product, with greater amount of growth factors and be capable of improvement the healing capacity in tissues with poor blood supply, such as tendons and joints, reducing the time of recovery of the animal. It is known that the growth factors are derived from platelets, but the positive correlation between the increase in platelet count and the higher concentration of growth factors is still a reason for discussion among the authors. In order to obtain a final product with a higher platelet count, it is frequent to increase the speed or number of centrifugations in the methodology employed, increasing the risk of early platelet aggregation. The aim of the present study is to verify the effect of double centrifugation in PRP preparation by comparing platelet counts, TGF-&#946;1 growth factor concentrations, and degree of platelet activation through percentage of aggregation. Twelve horses, male, aged 3 to 5 years-old, clinically healthy were subjected. Two different protocols were performed: one with single centrifugation and the other with double centrifugation. In the first one, the anticoagulated blood was centrifugated at 141G/12 minutes; while in the second one the first centrifugation was 300G/5 minutes followed by 700G/15 minutes, with rest between them and after. The products obtained after each centrifugation were submitted to platelet counting, aggregation test and measurement of TGF-&#946;1 by ELISA kit. The results showed a higher platelet concentration when double centrifugation protocol was used. The aggregometry test evidenced a greater activation of the platelets during the preparation of PRP when submitted to higher centrifugation velocities (times g), and not to double centrifugation. Quantification of TGF-&#946;1 showed no difference when performed on samples with only one centrifugation, but was higher values in the final product of the second centrifugation. The determination coefficient and Pearsons correlation coefficient showed a positive correlation between the platelet count and TGF-&#946;1 concentration. The double centrifugation protocol proved to be more effective at concentrating platelets and consequently higher amounts of TGF-&#946;1, not being impaired by early activation during obtainment.

Orientador: Andrigo Barboza De Nardi / Coorientador: Paola Castro Moraes / Banca: Rafael Ricardo Huppes / Banca: Jorge Luiz Costa Castro / Banca: Bruno Watanabe Minto / Banca: Pamela Rodrigues Reina Moreira / Resumo: Enxertos cutâneos são segmentos da epiderme e derme completamente removido da região doadora e transferidos para o local receptor, sem a presença de pedículo vascular. O curativo compressivo é recomendado por otimizar o contato do enxerto com o leito da ferida. O sucesso da cicatrização dos enxertos depende de adequada angiogênese, e por serem desprovidos de pedículo vascular o procedimento cirúrgico pode ser comprometido. Assim, para evitar complicações o plasma rico em plaquetas (PRP) é uma substância que estimula a angiogênese e auxilia na reparação tecidual. Sendo assim, delineou-se um estudo a fim de avaliar a eficácia do gel de PRP associado com esponjas cirúrgicas favorecerem a integração do enxerto ao leito receptor nos modelos de enxertos em camada e malha. Foi realizado no Hospital Veterinário da UNESP, câmpus de Jaboticabal - SP, um estudo com 64 coelhos, separados em oito grupos, com oito animais, todos submetidos à técnica de cirurgia reconstrutiva para confecção de enxerto. Os grupos foram compreendidos em: Gprpc (gel de PRP sem associação da esponja cirúrgica no enxerto de camada), Gprpce (gel de PRP associado com esponja cirúrgica como forma de curativo no enxerto de camada), Gcc (solução fisiológica 0,9% sem associação da esponja cirúrgica no enxerto de camada), Gcce (solução fisiológica 0,9% associada com esponjas cirúrgicas no enxerto de camada), Gprpm (gel de plasma PRP sem associação da esponja cirúrgica no enxerto de malha), Gprpme (gel de PRP associado ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Skin grafts are segments of the epidermis and dermis completely removed from the donor region and transferred to the receptor site without the presence of vascular pedicle. Pressure dressing after surgery is recommended to optimize the contact of the graft to the wound bed, and allow proper angiogenesis. The success of the healing of the grafts depends on proper angiogenesis, and being devoid of vascular pedicle the surgical procedure may be compromised. Thus, to avoid complications platelet rich plasma (PRP) is a substance that has been investigated for having angiogenesis factors which stimulate and aid in tissue repair. Thus, it outlined a study to assess the efficacy of PRP gel employment associated with surgical sponges to improve the integration of the graft in the recipient bed models grafts mesh and tier. It was held at the Veterinary Hospital UNESP, campus of Jaboticabal - SP, a study of 64 rabbits, divided into eight groups with eight animals, all underwent reconstructive surgery technique for making graft. The groups were comprised of: Gprpc received PRP gel unassociated surgical sponge in the tier graft, Gprpce received PRP gel associated with surgical sponge as a way of dressing in tier graft, Gcc received 0.9% saline without association surgical sponge in tier graft, Gcce received 0.9% saline solution associated with surgical sponges in the tier graft, Gprpm received plasma PRP gel unassociated surgical sponge in mesh graft, Gprpme received PRP gel associated with surgical sponge as way of dressing in mesh graft, Gcm receive 0.9% saline solution without associated surgical sponge in mesh graft and Gcce receive 0.9% saline solution associated with surgical sponges in mesh graft. Evaluations were performed macroscopic lesions after three, seven, and 14 days of surgery. To evaluate the wound healing took place microscopic ev... (Complete abstract click electronic access below) / Doutor

Orientador: Maria José Hitomi Nagata / Banca: Valdir Gouveia Garcia / Banca: Alvaro Francisco Bosco / Banca: Joni Augusto Cirelli / Banca: Luciana Saraiva / Resumo: Este estudo avaliou a influência do plasma rico em plaquetas derivado do aspirado de medula óssea (PRP-bma) na cicatrização de defeitos de fenestração periodontal (PFD) em ratos. PFD foram criados cirurgicamente na mandíbula de 40 ratos. Os animais foram divididos randomicamente em 2 grupos: C e PRP-bma - os defeitos foram preenchidos com coágulo sanguíneo ou PRP-bma, respectivamente. Os animais foram submetidos à eutanásia aos 10 ou 30 dias pós-operatórios. Foram realizadas análises histológica, histométrica e imunoistoquímica. A porcentagem de novo osso (NB), densidade do novo osso formado (DNB), novo cemento (NC) e extensão do defeito remanescente (ERD) foram avaliados histometricamente. Foram realizadas reações imunoistoquímicas para detecção de antígeno nuclear de proliferação celular (PCNA), sialoproteína óssea (BSP), osteocalcina (OCN) e fosfatase ácida resistente ao tartarato (TRAP). Células imunomarcadas foram quantificadas. Os dados foram analisados estatisticamente (ANOVA; Tukey, P < 0,05). Aos 10 dias, os grupos C e PRP-bma apresentaram quantidades similares de NB e DNB, e não foi observada formação de NC. Aos 30 dias, os grupos C e PRP-bma apresentaram quantidades similares de NB e DNB; o Grupo PRP-bma mostrou significativa formação de NC, com fibras colágenas inseridas obliquamente ou perpendicularmente à superfície radicular, sendo que nenhum espécime do Grupo C apresentou formação de NC. O Grupo PRP-bma apresentou um número significativamente maior de células PCNA-postivas (aos 10 dias) e BSP-positivas (aos 10 e 30 dias) que o Grupo C. Não foram observadas diferenças significativas no número de células OCN-positivas ou TRAP-positivas entre os grupos aos 10 ou 30 dias. Pode-se concluir que o PRP-bma promoveu formação de NC com ligamento periodontal funcional / Abstract: This study evaluated the influence of platelet-rich plasma derived from bone marrow aspirate (PRP-bma) on the healing of periodontal fenestration defects (PFD) in rats. PFD were surgically created in the mandible of 40 rats. The animals were randomly divided into 2 groups: C and PRP-bma - defects were filled with blood clot or PRP-bma, respectively. Animals were euthanized at either 10 or 30 days post-operative. Histologic, histometric and immunohistochemical analyses were performed. Percentage of new bone (NB), density of newly formed bone (DNB), new cementum (NC) and extension of remaining defect (ERD) were histometrically evaluated. Proliferating cell nuclear antigen (PCNA), bone sialoprotein (BSP), osteocalcin (OCN), and tartrate-resistant acid phosphatase (TRAP) immunohistochemical staining were performed. Immunolabeled cells were quantified. Data were statistically analyzed (ANOVA; Tukey, P < 0.05). At 10 days, groups C and PRP-bma presented similar amounts of NB and DNB, NC formation was not observed. At 30 days, groups C and PRP-bma presented similar amounts of NB and DNB; Group PRP-bma showed significant NC formation, with collagen fibers inserted obliquely or perpendicularly to the root surface, while NC formation was not observed in any Group C specimen. Group PRP-bma presented a significantly higher number of PCNA-positive (at 10 days) and BSP-positive cells (at 10 and 30 days) than Group C. No significant differences in the number of either OCN-positive or TRAP-positive cells were observed between groups at 10 or 30 days. It can be concluded that PRP-bma promoted NC formation with a functional periodontal ligament / Doutor