Skin graft associated with platelet-rich plasma in correcting extensive injuries resulting from the resection of skin cancer chemically induced in rats

ABSTRACT Purpose: To evaluate whether using platelet-rich plasma (PRP) in the graft recipient bed after the resection of a neoplasia can influence its recurrence because this product stimulates angiogenesis, mitogenesis and chemotaxis. Methods: A study with 30 rats Wistar (Rattus norvegicus albinus), which were separated into group A (induction of carcinogenesis, PRP in the postoperative period) and group B (induction of carcinogenesis, absence of PRP in the postoperative period), with 15 animals in each. Carcinogenesis was induced on the skin of the animals' chest by the topical application of 0.5% dimethylbenzantracene (DMBA) diluted in acetone. After surgical resection of the induced neoplasia, PRP was used to stimulate angiogenesis before surgical wound synthesis. Data on the control and experimental groups and macroscopic and microscopic variables were evaluated using analysis of variance and the Tukey's test (5%). Results: It was possible to determine that the use of PRP is good in reconstructive surgeries, but it is contraindicated in patients during tumor resection, as it can cause changes in the surgical bed, in addition to stimulating recurrences and metastases. Conclusions: PRP may interact with tumour cells that were in the recipient site of the surgical wound during the resection of a neoplasia, and a local recurrence process can be triggered by applying this product.

In-vitro production and pre-validation of lyophilized canine platelet-rich plasma for therapeutic use

ABSTRACT: Platelet-rich plasma (PRP) has been considered a promising therapeutic alternative, since platelets are rich in growth factors that are used in the Regenerative Medicine field. However, fresh PRP cannot be stored for long periods. This study aimed to develop a protocol for obtaining lyophilized canine PRP capable of maintaining viability after its reconstitution. For that purpose, canine PRP extraction and lyophilization protocols were initially tested. Subsequently, assays were carried out to quantify the growth factors VEGF and TGF-, before and after the lyophilization process, gelation test and the three-dimensional gel structure analysis of the reconstituted lyophilized PRP by electron microscopy, as well as in vitro cell proliferation test in lyophilized PRP gel. Additionally, the immunogenicity test was performed, using allogeneic samples of lyophilized PRP. The results showed that the lyophilized PRP had adequate therapeutic concentrations of growth factors VEGF and TGF- (9.1pg/mL and 6161.6pg/mL, respectively). The reconstituted PRP gel after lyophilization showed an in vitro durability of 10 days. Its electron microscopy structure was similar to that of fresh PRP. In the cell proliferation test, an intense division process was verified in mesenchymal stem cells (MSCs) through the three-dimensional mesh structure of the lyophilized PRP gel. The immunogenicity test showed no evidence of an immune reaction. The findings were promising, suggesting the possibility of having a lyophilized canine PRP that can be marketed. New in vivo and in vitro studies must be carried out for therapeutic confirmation.

RESUMO: O plasma rico em plaquetas (PRP) é uma alternativa terapêutica promissora, pois as plaquetas são ricas em fatores de crescimento com ação na regeneração de tecidos. No entanto, o PRP fresco não pode ser armazenado por longos períodos. Esse trabalho teve como objetivo desenvolver um protocolo de obtenção de PRP liofilizado canino capaz de manter a viabilidade pós reconstituição. Portanto, foram testados diversos protocolos de extração e liofilização. Para validação do PRP canino liofilizado foi analisada a concentração dos fatores de crescimento VEGF e TGF- antes e após o processo de liofilização, a estrutura tridimensional do PRP liofilizado reconstituído em forma de gel por microscopia eletrônica e seu efeito in vitro na proliferação de células-tronco mesenquimais. Os resultados demonstraram que o PRP liofilizado apresentou concentrações terapêuticas adequadas dos fatores de crescimento VEGF e TGF- (9,1pg/ml e 6161,6pg/ml, respectivamente). O gel de PRP reconstituído após liofilização apresentou uma durabilidade in vitro de 10 dias, sua estrutura tridimensional mostrou-se semelhante ao PRP fresco e proporcionou intensa proliferação de células-tronco mesenquimais durante o cultivo. O teste de imunogenicidade não demonstrou evidências de reação imune. Os achados foram promissores, sugerindo a possibilidade de uso de PRP canino liofilizado para o mercado. Novos estudos in vivo e in vitro deverão ser conduzidos para comprovação terapêutica.

In-vitro production and pre-validation of lyophilized canine platelet-rich plasma for therapeutic use / Produção e validação de plasma rico em plaquetas canino liofilizado para fins de utilização terapêutica

Platelet-rich plasma (PRP) has been considered a promising therapeutic alternative, since platelets are rich in growth factors that are used in the Regenerative Medicine field. However, fresh PRP cannot be stored for long periods. This study aimed to develop a protocol for obtaining lyophilized canine PRP capable of maintaining viability after its reconstitution. For that purpose, canine PRP extraction and lyophilization protocols were initially tested. Subsequently, assays were carried out to quantify the growth factors VEGF and TGF-β, before and after the lyophilization process, gelation test and the three-dimensional gel structure analysis of the reconstituted lyophilized PRP by electron microscopy, as well as in vitro cell proliferation test in lyophilized PRP gel. Additionally, the immunogenicity test was performed, using allogeneic samples of lyophilized PRP. The results showed that the lyophilized PRP had adequate therapeutic concentrations of growth factors VEGF and TGF-β (9.1pg/mL and 6161.6pg/mL, respectively). The reconstituted PRP gel after lyophilization showed an in vitro durability of 10 days. Its electron microscopy structure was similar to that of fresh PRP. In the cell proliferation test, an intense division process was verified in mesenchymal stem cells (MSCs) through the three-dimensional mesh structure of the lyophilized PRP gel. The immunogenicity test showed no evidence of an immune reaction. The findings were promising, suggesting the possibility of having a lyophilized canine PRP that can be marketed. New in vivo and in vitro studies must be carried out for therapeutic confirmation.(AU)

O plasma rico em plaquetas (PRP) é uma alternativa terapêutica promissora, pois as plaquetas são ricas em fatores de crescimento com ação na regeneração de tecidos. No entanto, o PRP fresco não pode ser armazenado por longos períodos. Esse trabalho teve como objetivo desenvolver um protocolo de obtenção de PRP liofilizado canino capaz de manter a viabilidade pós reconstituição. Portanto, foram testados diversos protocolos de extração e liofilização. Para validação do PRP canino liofilizado foi analisada a concentração dos fatores de crescimento VEGF e TGF-β antes e após o processo de liofilização, a estrutura tridimensional do PRP liofilizado reconstituído em forma de gel por microscopia eletrônica e seu efeito in vitro na proliferação de células-tronco mesenquimais. Os resultados demonstraram que o PRP liofilizado apresentou concentrações terapêuticas adequadas dos fatores de crescimento VEGF e TGF- β (9,1pg/ml e 6161,6pg/ml, respectivamente). O gel de PRP reconstituído após liofilização apresentou uma durabilidade in vitro de 10 dias, sua estrutura tridimensional mostrou-se semelhante ao PRP fresco e proporcionou intensa proliferação de células-tronco mesenquimais durante o cultivo. O teste de imunogenicidade não demonstrou evidências de reação imune. Os achados foram promissores, sugerindo a possibilidade de uso de PRP canino liofilizado para o mercado. Novos estudos in vivo e in vitro deverão ser conduzidos para comprovação terapêutica.(AU)

In-vitro production and pre-validation of lyophilized canine platelet-rich plasma for therapeutic use / Produção e validação de plasma rico em plaquetas canino liofilizado para fins de utilização terapêutica

Platelet-rich plasma (PRP) has been considered a promising therapeutic alternative, since platelets are rich in growth factors that are used in the Regenerative Medicine field. However, fresh PRP cannot be stored for long periods. This study aimed to develop a protocol for obtaining lyophilized canine PRP capable of maintaining viability after its reconstitution. For that purpose, canine PRP extraction and lyophilization protocols were initially tested. Subsequently, assays were carried out to quantify the growth factors VEGF and TGF-β, before and after the lyophilization process, gelation test and the three-dimensional gel structure analysis of the reconstituted lyophilized PRP by electron microscopy, as well as in vitro cell proliferation test in lyophilized PRP gel. Additionally, the immunogenicity test was performed, using allogeneic samples of lyophilized PRP. The results showed that the lyophilized PRP had adequate therapeutic concentrations of growth factors VEGF and TGF-β (9.1pg/mL and 6161.6pg/mL, respectively). The reconstituted PRP gel after lyophilization showed an in vitro durability of 10 days. Its electron microscopy structure was similar to that of fresh PRP. In the cell proliferation test, an intense division process was verified in mesenchymal stem cells (MSCs) through the three-dimensional mesh structure of the lyophilized PRP gel. The immunogenicity test showed no evidence of an immune reaction. The findings were promising, suggesting the possibility of having a lyophilized canine PRP that can be marketed. New in vivo and in vitro studies must be carried out for therapeutic confirmation.(AU)

O plasma rico em plaquetas (PRP) é uma alternativa terapêutica promissora, pois as plaquetas são ricas em fatores de crescimento com ação na regeneração de tecidos. No entanto, o PRP fresco não pode ser armazenado por longos períodos. Esse trabalho teve como objetivo desenvolver um protocolo de obtenção de PRP liofilizado canino capaz de manter a viabilidade pós reconstituição. Portanto, foram testados diversos protocolos de extração e liofilização. Para validação do PRP canino liofilizado foi analisada a concentração dos fatores de crescimento VEGF e TGF-β antes e após o processo de liofilização, a estrutura tridimensional do PRP liofilizado reconstituído em forma de gel por microscopia eletrônica e seu efeito in vitro na proliferação de células-tronco mesenquimais. Os resultados demonstraram que o PRP liofilizado apresentou concentrações terapêuticas adequadas dos fatores de crescimento VEGF e TGF- β (9,1pg/ml e 6161,6pg/ml, respectivamente). O gel de PRP reconstituído após liofilização apresentou uma durabilidade in vitro de 10 dias, sua estrutura tridimensional mostrou-se semelhante ao PRP fresco e proporcionou intensa proliferação de células-tronco mesenquimais durante o cultivo. O teste de imunogenicidade não demonstrou evidências de reação imune. Os achados foram promissores, sugerindo a possibilidade de uso de PRP canino liofilizado para o mercado. Novos estudos in vivo e in vitro deverão ser conduzidos para comprovação terapêutica.(AU)

Allogenic platelet-rich plasma in induced ulcers in rat's cornea / [Plasma rico em plaquetas alogênico em úlceras de córnea induzidas em ratos]

The objective in this study was to evaluate the clinic effect of applying allogenic platelet-rich plasma (PRP) heated or not, for treating cornea ulcers, including the dosage of PDGF-BB in the cornea. The ulcers were induced, standardizing the left eye from 81 rats (Ratus norvegicus, albinus variety), assigned randomly into three groups (N=27): control group (CG) which did not receive any topic treatment; heated PRP group (GA) and PRP group (GP), which received topical treatment every eight hours for five days. Each group underwent evaluation at 24 hours (M1), three days (M3) and five days (M5). The clinical exam evaluated the opacity, vascularization and corneal repair. The corneal PDGF-BB was dosed through the ELISA method. The corneal opacity was decreased in PRP-treated animals (GA and GP) and corneal repair time reduced when compared to CG at M1 and M5. Furthermore, GP showed greater vascularization at M3 compared to M1. Applied allogenic PRP eye drops, heated or not, speed up corneal healing, and reduce corneal repair time. However, the corneal PDGF concentration was not altered in any of the treatments.(AU)

Objetivou-se avaliar o efeito clínico da aplicação de plasma rico em plaquetas alogênico (PRP) aquecido ou não, no tratamento de úlceras de córnea, como a dosagem de PDGF-BB na córnea. As úlceras foram induzidas, padronizando-se o olho esquerdo de 81 ratos (Rattus norvegicus, variedade albinus), aleatoriamente, nos três grupos (N = 27): grupo controle (CG), que não recebeu nenhum tratamento tópico; grupo PRP aquecido (GA) e grupo PRP (GP), que receberam tratamento tópico a cada oito horas, durante cinco dias. Cada grupo foi subdividido em 24 horas (M1), três dias (M3) e cinco dias (M5). O exame clínico avaliou a opacidade, a vascularização e o reparo corneano. O PDGF-BB corneano foi dosado pelo método Elisa. Houve diminuição da opacidade da córnea nos animais tratados com PRP (GA e GP) e diminuição do tempo de reparo da córnea em comparação com CG, M1 e M5. Além disso, foi observada maior vascularização no GP no momento M3 em relação ao M1. A aplicação de colírios de PRP alogênico, aquecidos ou não, acelera a cicatrização da córnea, além de reduzir o tempo de reparo da córnea. No entanto, a concentração de PDGF na córnea não se alterou em nenhum dos tratamentos.(AU)

Effects of homologous and heterologous rich platelets plasma, compared to poor platelets plasma, on cutaneous healing of rabbits

Purpose: To evaluate and compare the effects of homologous and heterologous PRP (Platelet-Rich Plasma) on the quality and speed of skin wound healing, compared to Poor Platelet Plasma (PPP). Methods: Twenty-one male adult rabbits were used; two for preparing homologous PRP, with the rest of them separated randomly in three groups, according to the treatment received: PPP - control (n=5), homologous PRP (n=7), heterologous (n=7). Excisional skin wounds were made on the back of the animals, for the application of homologous and heterologous PPP and PRP. At the 14th post-operative day (POD), the animals were subjected to a new wound, and the treatments were inverted. The wounds were evaluated macroscopically and histologically. Results: A larger percentage of scar retraction was observed on the group treated with heterologous PRP, compared to homologous PRP, at the third POD, an increase of 25.03% (p=0.01). No other statistically significant differences among treatments were observed. Among every group, skin healing was efficient, without local adverse effects. Conclusions: Heterologous PRP contributed with more tissue retraction at the beginning of the wound healing process. After this, there were no differences on the wound healing skin process treated with PRP or PPP. However, our findings suggest the presence of others plasmatic factors, besides platelets, which could also contribute to the wound healing process, and thus, should be further investigated.(AU)

The influence of platelet-rich plasma (PRP) on colonic anastomosis healing impaired by intraperitoneal 5-flourouracil application: an experimental study

Purpose 5-flourourasil (5-FU) is commonly used for early intraperitoneal chemotherapy in colorectal or appendiceal cancer patients with peritoneal carcinomatosis. Due to its effect, anastomosis healing can be impaired and leads to anastomotic leakage. In this study, we aimed to investigate the potential healing effect of platelet-rich plasma (PRP) on colonic anastomosis impaired by intraperitoneal 5-flourouracil application. Methods After ten rats were sacrificed for preparing PRP, forty Wistar-albino rats were subjected to colonic anastomosis, and randomly allocated into four groups including 10 rats each. According to receiving PRP and/or 5-FU application, the groups were formed as control (C), 5-FU without PRP (CT), anastomosis with PRP (C-PRP), and 5-FU with PRP (CT-PRP). CT and CT-PRP groups also received 5-FU intraperitoneally on postoperative day 1 (POD 1). All animals were euthanized on pod 7. The body weight change, anastomotic bursting pressure (ABP), tissue hydroxiprolin (TH) and histopathological examination of each group were analyzed. Results 5-FU application significantly reduced ABP levels when compared with group C, C-PRP and CT-PRP (for each comparison, p 0,01). PRP application in CT-PRP group raised the measure of ABP up to the levels of C group. Although tissue hydroxyproline levels (THL) levels of CT-PRP group were found higher than CT group, it was not significant (p=0.112). Microscopically, comparing with CT group, PRP application significantly promoted the healing of colonic anastomosis subjected to 5-FU application by improving tissue edema, necrosis, submucosal bridging and collagen formation (p 0.05). Tissue healing in CT-PRP group was observed as good as the control groups. (C, C-PRP, p=0.181, p=0.134; respectively). Conclusion PRP administration on colonic anastomosis significantly promotes the healing process of anastomosis in rats receiving 5-FU. This result encourages further clinical use of PRP to reduce the frequency of AL in patients receiving EPIC.(AU)

Reconstruction of parietal bone defects with adiposederived mesenchymal stem cells: experimental study

Purpose: This study assessed the regeneration potential of mesenchymal stem cells (MSC) from adipose tissue associated with platelet-rich plasma (PRP) in bone regeneration. Methods: Thirty Wistar rats (Rattus norvegicus albinos) were divided into five groups (according to the grafting material and time to euthanasia): (1) autograft - 14 days (control), (2) autograft - 28 days (control), (3) MSC + PRP - 14 days, (4) MSC + PRP + papaverine - 14 days and (5) MSC + PRP + papaverine - 28 days. After euthanasia, the graft was removed and histological slides were prepared. They were assessed by a blinded pathologist using a previously published histological scale as parameter. Results: There was some degree of neoformed bone trabeculae (NBT) in 93.3% of the samples, as well as osteoblastic activity (OA). The autograft groups (14 and 28 days) had higher levels in the formation of bone trabeculae. Nonparametric data were analyzed using the Wilcoxon-Mann-Whitney test and proved not to be statistically significant at p 0.05. Conclusions: Experimental parietal bone reconstruction, combining MSC, PRP and papaverine presented regeneration in all groups with no significant difference among them.(AU)

Uso do plasma rico em plaquetas associado a colírios no tratamento de ceratite ulcerativa em equino / Use of rich plasma on platters associated with collieries in the treatment of ulcerative caeratitis in equine

A córnea é a primeira estrutura do olho a receber luz, sendo localizada na parte mais externado bulbo ocular. Devido a sua localização está sujeita, com maior frequência, a processos lesivos, como a úlcera de córnea, também chamada de ceratite ulcerativa. É a patologia com maior ocorrência dentre as afecções oftalmológicas em equinos, e caso não tenha o tratamento adequado, pode levar a perda da visão do animal. Diante desse contexto, objetivo do estudo foi relatar um caso clínico de ceratite ulcerativa em equino tratado com plasma rico em plaquetas (PRP) associado a colírios. Um equino, macho, de 3 anos de idade, raça Quarto de Milha, pelagem alazã foi encaminhado para atendimento veterinário por ter sofrido uma pancada em um caibro. Durante o exame clínico foi constatado que o cavalo apresentava quemose, epífora, blefaroespasmo e perda temporária da capacidade visual. Baseado nos sinais clínicos e histórico a suspeita clínica foi de ceratite ulcerativa. O tratamento indicado foi instilação de Ocucan® colírio, Ciprovet® e PRP, duas gotas de cada, duas vezes ao dia, além de lavagem com solução fisiológica. O animal apresentou evolução significativa ao tratamento e com 30 dias foi liberado para retornar a propriedade e as atividades físicas gradativamente. Dessa forma, podemos concluir que o uso de plasma rico em plaquetas constitui uma ferramenta importante e acessível no tratamento de ceratite ulcerativa em equinos, inclusive demonstrando bons resultados na caraterística da cicatrização e no tempo da mesma.

The cornea is the first structure of the eye receiving light, being located at the outermost part of the ocular bulb. Due to its location is subject, more often, the harmful processes such as corneal ulcer, also called keratitis Ulcerative. It is the disease with higher incidence among ophthalmologic diseases in horses, and if no proper treatment, can lead to the animal vision loss. In this context, objective of the study was to report a case of keratitis Ulcerative equinus treated with platelet-rich plasma (PRP) associated with eye drops. A horse, male, 3 years old, Quarter Horses, sorrel coat was referred to veterinary care after suffering a stroke on a rafter. During the clinical examination revealed that the horse had chemosis, epiphora blepharospasm and temporary loss of vision. Based on clinical signs and history clinical suspicion was keratitis Ulcerative. The treatment was Ocucan® instillation of eye drops, Ciprovet® PRP and two drops each, twice a day, and washing with saline solution. The animal showed evolution significant treatment and was released 30 days to return the property and physical activity gradually. Thus, we conclude that the use of platelet rich plasma is an important tool and accessible in the treatment of ulcerative keratitis in horses, including demonstrating good results in healing characteristic and time thereof.

Uso do plasma rico em plaquetas como adjuvante no tratamento de lesão meniscal e tendinose de calcâneo em um cão / Use of platelet-rich plasma as an adjuvant on treatment of meniscal lesion and tendinosis of calcaneus in a dog

O objetivo deste trabalho é relatar o caso de um cão com lesão meniscal associado à tendinose de calcâneo tratado com aplicação única de plasma rico em plaquetas (PRP) intraarticular e intratendínea. Foi atendido um cão da raça Boxer com dificuldade de deambulação há três meses. Observou-se claudicação grau II/IV em ambos os membros pélvicos, atrofia muscular bilateral, dor e leve crepitação em joelho esquerdo, discreta instabilidade aos testes de gaveta e tibial cranial, com intensa fibrose medial à articulação e espessamento/rigidez do tendão calcâneo direito. O estudo radiográfico demonstrou artrose moderada no joelho esquerdo e, ao exame ultrassonográfico, constatou-se áreas de hipoecogenicidade em menisco medial esquerdo e ruptura de fibras musculares do tendão calcâneo direito. Sendo assim, o paciente foi diagnosticado com insuficiência do ligamento cruzado cranial crônica, com estabilidade fibrótica periarticular, e lesão meniscal em joelho esquerdo e tendinose de calcâneo direito. Foram prescritos condroprotetores, diacereína, fisioterapia e uma aplicação de PRP intra-articular e intratendínea. Para a concentração do PRP, foram coletados 50mL de sangue autólogo, os quais foram depositados em frascos contendo citrato de sódio. Foi realizada dupla centrifugação, sendo a primeira com força de120g e a segunda 240g, ambas por dez minutos. Ao final, obteve-se 2mL de PRP com246.000 plaquetas/µL (antes da concentração o sangue tinha 141.000 plaquetas/µL). A aplicação ocorreu de forma asséptica, sob anestesia geral e guiada por ultrassom. Foram injetados 1ml intra-articular em joelho esquerdo, 0,25mL em região perilesional e 0,75mLintralesional em tendão calcâneo direito. Após 14 dias, notou-se grau de claudicação I/IV e discreto fortalecimento muscular. Foi mantido o tratamento clínico com fisioterapia e condroprotetores com retornos periódicos para acompanhamento da evolução. Conclui-se que o uso do PRP foi importante adjuvante na terapia clínica empregada.

The aim of this paper is to report the case of a dog with meniscal lesion associated with calcaneal tendinosis treated with single application of intra-articular and intratendinous platelet rich plasma (PRP). A dog of the Boxer racer with difficulty walking was treated three months ago. Grade II/IV lameness was observed in both pelvic limbs, bilateral muscle atrophy, pain and slight crepitation in the left knee, slight instability to the drawer and cranial tibial tests, with intense medial fibrosis to the joint and right calcaneal tendon thickening/stiffness. The radiographic study showed moderate arthrosis in the left knee, and ultrasound examination revealed areas of hypoechogenicity in the left medial meniscus and rupture of the right calcaneal tendon muscle fibers. Thus, the patient was diagnosed with chronic cranial cruciate ligament insufficiency, periarticular fibrotic stability, and left meniscal injury and right calcaneal tendinosis. Chondroprotectors, diacerein, physiotherapyand an application of intra-articular and intratendinous PRP were prescribed. For the PRP concentration, 50mL of autologous blood were collected and deposited in vials containing sodium citrate. A double centrifugation was performed, the first strength of 120g and the second 240g, both for ten minutes. At the end, 2mL of PRP was obtained with 246.000platelets/µL (before concentration the blood had 141.000 platelets/µL). The application occurred aseptically, under general anesthesia and guided by ultrasound. We injected 1mlintra-articular in the left knee, 0.25mL in the perilesional region and 0.75mL intralesional in the right calcaneal tendon. After 14 days, I/IV lameness and slight muscle strengthening were noted. Clinical treatment with physiotherapy and chondroprotectors was maintained, with periodic returns to follow the evolution. It was concluded that the use of PRP was an important adjuvant in the clinical therapy employed.

Associação de plasma rico em plaquetas de equino e pomada comercial no tratamento de feridas em pele de coelho / Rich plasma association in equine platform and commercial oil in writing of rabbit skin

Autologous platelet-rich plasma (PRP) is beneficial in the healing process of reconstructive surgeries, contributing to the stimulation of angiogenesis; however, heterologous plasma has been shown to be more effective. The objective of this study was to verify, by macro- and microscopic evaluation, whether PRP accelerates the healing process as compared to a commercial ointment containing gentamicin, sulfanilamide, sulfadiazine, urea, and vitamin A (study ointment). This study used 24 white New Zealand rabbits, aged 12 months, and each weighing approximately 3 kg. The animals were separated into 4 equal groups and underwent wound induction through skin removal in the thoracic (study wound) and lumbar (control wound) dorsal midline. Study wounds were co-treated with equine PRP and the study ointment. Control wounds were treated with only the study ointment. Group I underwent histological evaluation 3 days after the skin removal procedure, group II after 7 days, group III after 14 days, and group IV after 21 days. A skin fragment was collected from each animal for histological evaluation. The co-treatment with heterologous equine PRP and the study ointment accelerated the healing process in the surgically induced skin wounds, confirming the acceleratory effect of PRP on wound healing.

O plasma rico em plaquetas autólogo é importante no processo cicatricial de cirurgias reconstrutivas, auxiliando na estimulação da angiogênese; no entanto, o plasma heterólogo mostrou-se mais eficaz. Utilizou-se 24 coelhos da raça Nova Zelândia, com idade de 12 meses, com peso corporal cerca de 3,0 kg. Os animais foram separados em quatro grupos de igual número e submetidos a remoção de pele na linha média dorsal torácica (ferida tratada) e lombar (ferida controle). Nas tratadas, foi aplicado plasma rico em plaqueta de equino e pomada contendo gentamicina, sulfanilamida, sulfadiazina, ureia e vitamina A e, nas do grupo controle, somente a pomada. Nos do grupo I, foi coletado tecido cutâneo para a avaliação histológica com três dias de pós-operatório, nos do grupo II com sete dias, nos do grupo III com 14 dias e, nos do grupo IV, com 21 dias. Decorrido o período de avaliação de cada grupo, foi coletado fragmento de pele para avaliação histológica. O uso associado do plasma rico em plaquetas heterólogo de equino com a pomada contendo gentamicina, sulfanilamida, sulfadiazina, ureia e vitamina A, em feridas de pele induzidas cirurgicamente acelera o processo cicatricial.

Uso do plasma rico em plaquetas associado a colírios no tratamento de ceratite ulcerativa em equino / Use of rich plasma on platters associated with collieries in the treatment of ulcerative caeratitis in equine

A córnea é a primeira estrutura do olho a receber luz, sendo localizada na parte mais externado bulbo ocular. Devido a sua localização está sujeita, com maior frequência, a processos lesivos, como a úlcera de córnea, também chamada de ceratite ulcerativa. É a patologia com maior ocorrência dentre as afecções oftalmológicas em equinos, e caso não tenha o tratamento adequado, pode levar a perda da visão do animal. Diante desse contexto, objetivo do estudo foi relatar um caso clínico de ceratite ulcerativa em equino tratado com plasma rico em plaquetas (PRP) associado a colírios. Um equino, macho, de 3 anos de idade, raça Quarto de Milha, pelagem alazã foi encaminhado para atendimento veterinário por ter sofrido uma pancada em um caibro. Durante o exame clínico foi constatado que o cavalo apresentava quemose, epífora, blefaroespasmo e perda temporária da capacidade visual. Baseado nos sinais clínicos e histórico a suspeita clínica foi de ceratite ulcerativa. O tratamento indicado foi instilação de Ocucan® colírio, Ciprovet® e PRP, duas gotas de cada, duas vezes ao dia, além de lavagem com solução fisiológica. O animal apresentou evolução significativa ao tratamento e com 30 dias foi liberado para retornar a propriedade e as atividades físicas gradativamente. Dessa forma, podemos concluir que o uso de plasma rico em plaquetas constitui uma ferramenta importante e acessível no tratamento de ceratite ulcerativa em equinos, inclusive demonstrando bons resultados na caraterística da cicatrização e no tempo da mesma.(AU)

The cornea is the first structure of the eye receiving light, being located at the outermost part of the ocular bulb. Due to its location is subject, more often, the harmful processes such as corneal ulcer, also called keratitis Ulcerative. It is the disease with higher incidence among ophthalmologic diseases in horses, and if no proper treatment, can lead to the animal vision loss. In this context, objective of the study was to report a case of keratitis Ulcerative equinus treated with platelet-rich plasma (PRP) associated with eye drops. A horse, male, 3 years old, Quarter Horses, sorrel coat was referred to veterinary care after suffering a stroke on a rafter. During the clinical examination revealed that the horse had chemosis, epiphora blepharospasm and temporary loss of vision. Based on clinical signs and history clinical suspicion was keratitis Ulcerative. The treatment was Ocucan® instillation of eye drops, Ciprovet® PRP and two drops each, twice a day, and washing with saline solution. The animal showed evolution significant treatment and was released 30 days to return the property and physical activity gradually. Thus, we conclude that the use of platelet rich plasma is an important tool and accessible in the treatment of ulcerative keratitis in horses, including demonstrating good results in healing characteristic and time thereof.(AU)

Uso do plasma rico em plaquetas como adjuvante no tratamento de lesão meniscal e tendinose de calcâneo em um cão / Use of platelet-rich plasma as an adjuvant on treatment of meniscal lesion and tendinosis of calcaneus in a dog

O objetivo deste trabalho é relatar o caso de um cão com lesão meniscal associado à tendinose de calcâneo tratado com aplicação única de plasma rico em plaquetas (PRP) intraarticular e intratendínea. Foi atendido um cão da raça Boxer com dificuldade de deambulação há três meses. Observou-se claudicação grau II/IV em ambos os membros pélvicos, atrofia muscular bilateral, dor e leve crepitação em joelho esquerdo, discreta instabilidade aos testes de gaveta e tibial cranial, com intensa fibrose medial à articulação e espessamento/rigidez do tendão calcâneo direito. O estudo radiográfico demonstrou artrose moderada no joelho esquerdo e, ao exame ultrassonográfico, constatou-se áreas de hipoecogenicidade em menisco medial esquerdo e ruptura de fibras musculares do tendão calcâneo direito. Sendo assim, o paciente foi diagnosticado com insuficiência do ligamento cruzado cranial crônica, com estabilidade fibrótica periarticular, e lesão meniscal em joelho esquerdo e tendinose de calcâneo direito. Foram prescritos condroprotetores, diacereína, fisioterapia e uma aplicação de PRP intra-articular e intratendínea. Para a concentração do PRP, foram coletados 50mL de sangue autólogo, os quais foram depositados em frascos contendo citrato de sódio. Foi realizada dupla centrifugação, sendo a primeira com força de120g e a segunda 240g, ambas por dez minutos. Ao final, obteve-se 2mL de PRP com246.000 plaquetas/µL (antes da concentração o sangue tinha 141.000 plaquetas/µL). A aplicação ocorreu de forma asséptica, sob anestesia geral e guiada por ultrassom. Foram injetados 1ml intra-articular em joelho esquerdo, 0,25mL em região perilesional e 0,75mLintralesional em tendão calcâneo direito. Após 14 dias, notou-se grau de claudicação I/IV e discreto fortalecimento muscular. Foi mantido o tratamento clínico com fisioterapia e condroprotetores com retornos periódicos para acompanhamento da evolução. Conclui-se que o uso do PRP foi importante adjuvante na terapia clínica empregada.(AU)

The aim of this paper is to report the case of a dog with meniscal lesion associated with calcaneal tendinosis treated with single application of intra-articular and intratendinous platelet rich plasma (PRP). A dog of the Boxer racer with difficulty walking was treated three months ago. Grade II/IV lameness was observed in both pelvic limbs, bilateral muscle atrophy, pain and slight crepitation in the left knee, slight instability to the drawer and cranial tibial tests, with intense medial fibrosis to the joint and right calcaneal tendon thickening/stiffness. The radiographic study showed moderate arthrosis in the left knee, and ultrasound examination revealed areas of hypoechogenicity in the left medial meniscus and rupture of the right calcaneal tendon muscle fibers. Thus, the patient was diagnosed with chronic cranial cruciate ligament insufficiency, periarticular fibrotic stability, and left meniscal injury and right calcaneal tendinosis. Chondroprotectors, diacerein, physiotherapyand an application of intra-articular and intratendinous PRP were prescribed. For the PRP concentration, 50mL of autologous blood were collected and deposited in vials containing sodium citrate. A double centrifugation was performed, the first strength of 120g and the second 240g, both for ten minutes. At the end, 2mL of PRP was obtained with 246.000platelets/µL (before concentration the blood had 141.000 platelets/µL). The application occurred aseptically, under general anesthesia and guided by ultrasound. We injected 1mlintra-articular in the left knee, 0.25mL in the perilesional region and 0.75mL intralesional in the right calcaneal tendon. After 14 days, I/IV lameness and slight muscle strengthening were noted. Clinical treatment with physiotherapy and chondroprotectors was maintained, with periodic returns to follow the evolution. It was concluded that the use of PRP was an important adjuvant in the clinical therapy employed.(AU)

Associação de plasma rico em plaquetas de equino e pomada comercial no tratamento de feridas em pele de coelho / Rich plasma association in equine platform and commercial oil in writing of rabbit skin

Autologous platelet-rich plasma (PRP) is beneficial in the healing process of reconstructive surgeries, contributing to the stimulation of angiogenesis; however, heterologous plasma has been shown to be more effective. The objective of this study was to verify, by macro- and microscopic evaluation, whether PRP accelerates the healing process as compared to a commercial ointment containing gentamicin, sulfanilamide, sulfadiazine, urea, and vitamin A (study ointment). This study used 24 white New Zealand rabbits, aged 12 months, and each weighing approximately 3 kg. The animals were separated into 4 equal groups and underwent wound induction through skin removal in the thoracic (study wound) and lumbar (control wound) dorsal midline. Study wounds were co-treated with equine PRP and the study ointment. Control wounds were treated with only the study ointment. Group I underwent histological evaluation 3 days after the skin removal procedure, group II after 7 days, group III after 14 days, and group IV after 21 days. A skin fragment was collected from each animal for histological evaluation. The co-treatment with heterologous equine PRP and the study ointment accelerated the healing process in the surgically induced skin wounds, confirming the acceleratory effect of PRP on wound healing.(AU)

O plasma rico em plaquetas autólogo é importante no processo cicatricial de cirurgias reconstrutivas, auxiliando na estimulação da angiogênese; no entanto, o plasma heterólogo mostrou-se mais eficaz. Utilizou-se 24 coelhos da raça Nova Zelândia, com idade de 12 meses, com peso corporal cerca de 3,0 kg. Os animais foram separados em quatro grupos de igual número e submetidos a remoção de pele na linha média dorsal torácica (ferida tratada) e lombar (ferida controle). Nas tratadas, foi aplicado plasma rico em plaqueta de equino e pomada contendo gentamicina, sulfanilamida, sulfadiazina, ureia e vitamina A e, nas do grupo controle, somente a pomada. Nos do grupo I, foi coletado tecido cutâneo para a avaliação histológica com três dias de pós-operatório, nos do grupo II com sete dias, nos do grupo III com 14 dias e, nos do grupo IV, com 21 dias. Decorrido o período de avaliação de cada grupo, foi coletado fragmento de pele para avaliação histológica. O uso associado do plasma rico em plaquetas heterólogo de equino com a pomada contendo gentamicina, sulfanilamida, sulfadiazina, ureia e vitamina A, em feridas de pele induzidas cirurgicamente acelera o processo cicatricial.(AU)

Comparison of the protocols for obtaining platelet-rich plasma in dogs: a cellular study / Comparação entre protocolos para obtenção de plasma rico em plaquetas em cães: estudo celular

This study aimed to evaluate two protocols (PA and PB) that are used to obtain canine platelet-rich plasma (PRP) for cellularity. Twenty healthy dogs were used. Blood samples were collected and placed in two tubes of 3.2% sodium citrate. PA used double centrifugation at 210 x g, and 370 x g and PB used double centrifugation with 140 x g and 330 x g. The PRP samples from the protocols were examined in terms of their platelet, erythrocyte, and leukocyte count in the Neubauer chamber, differential leukocyte count and platelet morphological observation in blood smears. Data (mean and standard deviation) were analyzed with the 95% probability t-test (P <0.05) using Pearsons correlation to test the relationship between platelets and erythrocytes, platelets, and leukocytes, and the leukocyte count versus the erythrocytes. Very weak negative correlation between platelets and leukocytes (p= -0.03), weak negative correlation between platelets and erythrocytes (p= -0.3) and a strong positive correlation between leukocytes and erythrocytes (ρ = 0.75) were noted. Although, BP did not reach the desired mean of one million platelets (979300 ± 79631 cells / μL), both protocols, A and B (4.42 ± 1.61 and 3.85 ± 1.55 times more platelets than total blood, respectively) (p <0.05) were efficient in concentrating platelets. Platelet activation was present in 26.55 ± 6.72% of the PA platelets and 26.25 ± 7.03% in PB (p> 0.05). PA and PB presented low erythrocyte concentration (p> 0.05), and PA had more leukocytes (p <0.05) than PB, with higher concentrations of basophils that were segmented, and lymphocytes.(AU)

Foi proposto avaliar dois protocolos (PA e PB) para obtenção de plasma rico em plaquetas (PRP) canino quanto a celularidade. Foram utilizados 20 cães sadios e coletadas amostras sanguíneas, sendo acondicionados em dois tubos de citrato de sódio a 3,2%. O PA utilizando centrifugação dupla com 210 xG e 370 xG e PB utilizando centrifugação dupla com 140 xG e 330 xG. Amostras de PRP dos protocolos foram destinadas a contagem plaquetária, eritrocitária e leucocitária em câmara de Neubauer, contagem diferencial leucocitária e observação morfológica plaquetária em esfregaços sanguíneos. Analisou-se os dados (médias e desvios padrão) pelo Teste t com 95% de probabilidade (p<0,05) utilizando-se correlação de Pearson para testar a relação entre a contagem de plaquetas e eritrócitos, plaquetas e leucócitos e leucócitos em relação aos eritrócitos. Houve correlação negativa muito fraca entre plaquetas e leucócitos (ρ= -0,03), negativa fraca entre plaquetas e eritrócitos (ρ= -0,3) e correlação positiva forte entre leucócitos e eritrócitos (ρ=0,75). Embora o PB não tenha alcançando a média de um milhão de plaquetas desejado (979300 ± 79631 células/µL), ambos os protocolos, A e B (4,42 ± 1,61 e 3,85 ± 1,55 vezes mais plaquetas que o sangue total, respectivamente) (p<0,05), foram eficientes em concentrar plaquetas. A ativação plaquetária esteve presente em 26,55 ± 6,72 % das plaquetas do PA e 26,25 ± 7,03 % nas do PB (p>0,05). PA e PB apresentaram baixa concentração eritrocitária (p>0,05) e PA apresentou mais leucócitos (p<0,05) que PB, com maiores concentrações de basófilos, segmentados e linfócitos.(AU)

Evaluation of platelet-rich plasma gel as an angiogenesis-inducing agent in canine advancement skin flaps / Avaliação do gel de plasma rico em plaquetas como agente indutor da angiogênese em flapes cutâneos de avanço em cães

This work aimed to evaluate the effect of platelet-rich plasma (PRP) on advancement skin flaps in dogs regarding improvement of vascularization, with focus on increasing its viable area, since there are reports that it is a potential angiogenesis stimulator. The experimental group was composed of eight adult bitches, in which two advancement skin flaps were made in the ventral abdominal region. No product was applied in the control flap (CF), while PRP was used in the contralateral flap, called treated flap (TF). The areas were clinically evaluated every two days until the 7th postoperative day regarding skin color and presence of necrosis. At 10 days, both flaps were removed and submitted to histological examination and blood vessel morphometry. The vessels counted in each group were statistically analyzed by the F-test at 1% probability. Results showed no significant difference in macroscopic changes in the wound, or CF and TF vascularization, thus suggesting that PRP gel did not improve advancement skin flap angiogenesis in bitches under the experimental conditions in which this research was developed.(AU)

Objetivou-se com o presente artigo avaliar a ação angiogênica do gel de plasma rico em plaquetas (PRP) em flapes cutâneos de avanço em animais da espécie canina, visando aumentar a viabilidade da pele após o procedimento, uma vez que existem relatos de que o produto é um potente estimulador da angiogênese. O grupo experimental foi composto por oito cadelas adultas, onde foram confeccionados dois flapes de avanço (de padrão subdérmico) na região abdominal ventral. Em um dos flapes, considerado controle (FC) não foi aplicado nenhum produto, enquanto que no flape contralateral, denominado tratado (FT), foi usado o gel de PRP. As áreas foram macroscopicamente avaliadas a cada dois dias até o 7º dia de pós-operatório em relação à coloração da pele e presença de área de necrose, e com 10 dias ambos os flapes foram coletados por biópsia e submetidos ao exame histológico e morfometria dos vasos sanguíneos. Os vasos contados em cada grupo foram estatisticamente analisados pelo teste de F ao nível de 1% de probabilidades. Os resultados demonstraram que não houve diferença significativa nas alterações macroscópicas das feridas e na morfometria vascular dos FC e FT, sugerindo dessa maneira que dentro das condições experimentais nas quais a pesquisa foi executada, que o gel de PRP não incrementou a angiogênese de flapes de avanço em cadelas.(AU)

Evaluation of platelet-rich plasma gel as an angiogenesis-inducing agent in canine advancement skin flaps / Avaliação do gel de plasma rico em plaquetas como agente indutor da angiogênese em flapes cutâneos de avanço em cães

This work aimed to evaluate the effect of platelet-rich plasma (PRP) on advancement skin flaps in dogs regarding improvement of vascularization, with focus on increasing its viable area, since there are reports that it is a potential angiogenesis stimulator. The experimental group was composed of eight adult bitches, in which two advancement skin flaps were made in the ventral abdominal region. No product was applied in the control flap (CF), while PRP was used in the contralateral flap, called treated flap (TF). The areas were clinically evaluated every two days until the 7th postoperative day regarding skin color and presence of necrosis. At 10 days, both flaps were removed and submitted to histological examination and blood vessel morphometry. The vessels counted in each group were statistically analyzed by the F-test at 1% probability. Results showed no significant difference in macroscopic changes in the wound, or CF and TF vascularization, thus suggesting that PRP gel did not improve advancement skin flap angiogenesis in bitches under the experimental conditions in which this research was developed.(AU)

Objetivou-se com o presente artigo avaliar a ação angiogênica do gel de plasma rico em plaquetas (PRP) em flapes cutâneos de avanço em animais da espécie canina, visando aumentar a viabilidade da pele após o procedimento, uma vez que existem relatos de que o produto é um potente estimulador da angiogênese. O grupo experimental foi composto por oito cadelas adultas, onde foram confeccionados dois flapes de avanço (de padrão subdérmico) na região abdominal ventral. Em um dos flapes, considerado controle (FC) não foi aplicado nenhum produto, enquanto que no flape contralateral, denominado tratado (FT), foi usado o gel de PRP. As áreas foram macroscopicamente avaliadas a cada dois dias até o 7º dia de pós-operatório em relação à coloração da pele e presença de área de necrose, e com 10 dias ambos os flapes foram coletados por biópsia e submetidos ao exame histológico e morfometria dos vasos sanguíneos. Os vasos contados em cada grupo foram estatisticamente analisados pelo teste de F ao nível de 1% de probabilidades. Os resultados demonstraram que não houve diferença significativa nas alterações macroscópicas das feridas e na morfometria vascular dos FC e FT, sugerindo dessa maneira que dentro das condições experimentais nas quais a pesquisa foi executada, que o gel de PRP não incrementou a angiogênese de flapes de avanço em cadelas.(AU)

Platelet-rich plasma (PRP) therapy: An approach in reproductive medicine based on successful animal models

Platelet-rich plasma (PRP) has been fully studied for different clinical applications in veterinary medicine for many years with promising results. As a result, therapeutic studies to elucidate pathways for PRP use in human reproduction have been performed. PRP applications in human reproductive medicine are recent, but the role of platelet growth factors in improving the endometrial environment is well known. Indications for PRP therapy show its positive effects in promoting endometrial and follicular growth and gestation in assisted reproduction cycles, as has been proven in animals. We summarized the putative role of PRP on endometrial receptivity with a brief history of promising results in research and clinical therapies.

Platelet-rich plasma (PRP) therapy: An approach in reproductive medicine based on successful animal models

Platelet-rich plasma (PRP) has been fully studied for different clinical applications in veterinary medicine for many years with promising results. As a result, therapeutic studies to elucidate pathways for PRP use in human reproduction have been performed. PRP applications in human reproductive medicine are recent, but the role of platelet growth factors in improving the endometrial environment is well known. Indications for PRP therapy show its positive effects in promoting endometrial and follicular growth and gestation in assisted reproduction cycles, as has been proven in animals. We summarized the putative role of PRP on endometrial receptivity with a brief history of promising results in research and clinical therapies.(AU)

Utilização de plasma rico em plaquetas no tratamento de ferida cutânea em cão: relato de caso / Use of platelet-rich plasma in the treatment of cutaneous wound in dogs: a case report / Utilización de plasma rico en plaquetas en el tratamiento de herida cutánea en perro: relato de caso

O plasma rico em plaquetas (PRP) consiste em um concentrado suprafisiológico de plaquetas extraído a partir do sangue autógeno do paciente e centrifugado adequadamente. O presente relato descreve a utilização do PRP no tratamento de ferida cutânea, após exérese de massa com evolução de 4 meses em um cão fêmea, da raça Beagle, de 7 anos de idade. Citologia aspirativa por agulha fina (CAAF) foi realizada na massa e diagnóstico sugestivo de neoplasia de anexo cutâneo de baixo grau foi identificado. O tratamento instituído consistiu na excisão da massa e aplicações tópicas e intradérmicas do produto obtido nos bordos da ferida. A lesão apresentou diminuição da resposta inflamatória, cicatrização limpa, bordos uniformes, e ausência de exsudato purulento, tecido de granulação exagerado e complicações sob o caráter neoplásico da etiologia da ferida, cicatrizando completamente após 17 dias.(AU)

Platelet-rich plasma (PRP) consists of a supra-physiologic platelet concentrate extracted from the patient's autogenous blood, after being properly centrifuged. This report describes the use of PRP in the treatment of cutaneous wound, after excision of mass with 4-month evolution in a 7-year-old beagle bitch. Fine needle aspiration cytology (FNAC) was performed in the mass and suggestive diagnosis of cutaneous adnexal neoplasm was identified. The treatment consisted of mass excision, added by topical and intradermal applications of the studied product in the wound edges. The wound presented decreased inflammatory response, clean healing, uniform edges, and absence of purulent exudate, exaggerated granulation tissue or complications under the neoplastic character of the wound, healing completely after 17 days.(AU)

El plasma rico en plaquetas (PRP) consiste en un concentrado supra fisiológico de plaquetas extraído a partir de la sangre autógena del paciente y centrifugado adecuadamente. El presente relato describe la utilización del PRP en el tratamiento de herida cutánea, tras exéresis de masa con evolución de 4 meses en una perra, de la raza Beagle, de 7 años de edad. Citología aspirativa por aguja fina (CAAF), se realizó en la masa y el diagnóstico sugestivo de neoplasia de anexo cutáneo de bajo grado fue identificado. El tratamiento instituido consistió en la escisión de la masa y aplicaciones tópicas e intradérmicas en los bordes de la herida con el producto obtenido. La lesión presentó reducción de la respuesta inflamatoria, cicatrización limpia, bordes uniformes, ausencia de exudado purulento, tejido de granulación exagerado y complicaciones bajo el carácter neoplásico de la etiología de la herida, cicatrizando completamente después de 17 días.(AU)