Padronização do processamento do plasma rico em plaquetas e lisado de plaquetas para uso clínico
Arquivos
Data
2022-10-04
Autores
Nani, Luiz Augusto Silva [UNIFESP]
Orientadores
Nogueira, Adriana Luckow Invitti [UNIFESP]
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
O plasma rico em plaquetas (PRP) é um derivado do sangue total com alta
concentração de plaquetas e, portanto, fonte de fatores biológicos fundamentais para
processos de regeneração e reparo tecidual, cicatrização, modulação da resposta
inflamatória, mobilização e proliferação de células-tronco teciduais e neoangiogênese. Essas características fizeram com que seja largamente utilizado em estudos para verificação de sua segurança e eficácia para as mais diversas afecções, com destaque para as ortopédicas, ginecológicas e dermatológicas. Apesar disso, não existe padronização do preparo do PRP entre esses estudos, o que faz com que os resultados sejam variáveis e não comparáveis. Além disso, um subproduto do PRP, o lisado de plaquetas (PL) também possui grande potencial de aplicação clínica. Sendo assim, realizamos uma busca na literatura (de 2020 até 2022) e selecionamos os 50 artigos mais relevantes que fizeram uso clínico do PRP. Compilamos e comparamos os dados de metodologia de preparo e tipo de estudo. Experimentalmente, coletamos 10 amostras de sangue periférico que foram divididas e processadas de 3 maneiras diferentes: PRP leve (≤1000g), PRP alto (>1000g) e lisado plaquetário autólogo (PLA); além de 10 amostras de concentrados de plaquetas provenientes de doadores do banco de sangue que foram processadas em lisado plaquetário heterólogo (PLH). Os preparos foram padronizados e os produtos obtidos caracterizados por contagem, recuperação e atividade de plaquetas, atividade do Fator VIII e detecção de 40 fatores biológicos. Como esperado, o número de plaquetas e a recuperação aumentaram significativamente (p <0,001) no preparo de centrifugação alta (72,03 ± 4,62 %) quando comparado com a leve (44,58 ± 2,14 %). Entretanto, nenhuma diferença significativa foi observada na atividade plaquetária e do Fator VIII para os métodos testados. A força centrífuga também não interferiu na concentração de fatores solúveis nos grupos testados. O padrão de expressão do PLA foi semelhante ao PRP leve e alto dos mesmos voluntários,
enquanto que o PLH apresentou concentrações diferentes. Os métodos de preparo de PRP testados não interferem na atividade fisiológica das plaquetas, mas podem
interferir na disponibilidade e concentração de fatores solúveis. Isso justificaria a
alteração do método de preparo dependendo da aplicação clínica pretendida.
The platelet rich plasma (PRP) is a blood product that contains higher platelets concentration than the whole blood; being a rich source of biological factors involved in tissue regeneration and repair, wound healing, inflammatory response modulation, tissue stem cells mobilization and proliferation and neoangiogenesis. According to these properties, the PRP have been extensively explored in clinical trials to the most diverse applications mainly orthopedics, gynecological and dermatological conditions. Besides, there is a PRP product, the human platelet lysate (PL), that have the same potential application. Although the clinical results are very controversial as the PRP preparation is not standardized between the studies. According to this, we searched the literature from 2020 to 2022 for clinical trials applying PRP and selected the 50 most relevant ones, which were scrutinized and the various features of the described protocols were compiled. Furthermore, we collected 10 whole blood samples that were split and processed into three protocols: PRP mild (≤1,000g), PRP high (>1,000g) and autologous platelet lysate (PLA). Also, we prepared platelet lysate heterologous (PLH) from 10 blood from donors. The protocols were evaluated for platelet counting, recovery and activity, Factor VIII activity and detection of a 40 biological factors. The platelet number and recovery were significantly higher (p <0,001) in the high protocol (72.03 ± 4.62 %) when compared to the mild one (44.58 ± 2.14 %). However, no significant difference was observed on platelet activity and Factor VIII comparing the protocols. PRP preparation protocols also did not interfere on expression pattern, where the main differences observed where in the comparisons of pooled and individual samples. Samples from the same donor were grouped together in spite of the preparation protocol. The PRP processing method did not affect the platelet physiological activity but can have effect on the biological factors availability and concentration. It can justify the choice of PRP processing depending on the pretended clinical application.
The platelet rich plasma (PRP) is a blood product that contains higher platelets concentration than the whole blood; being a rich source of biological factors involved in tissue regeneration and repair, wound healing, inflammatory response modulation, tissue stem cells mobilization and proliferation and neoangiogenesis. According to these properties, the PRP have been extensively explored in clinical trials to the most diverse applications mainly orthopedics, gynecological and dermatological conditions. Besides, there is a PRP product, the human platelet lysate (PL), that have the same potential application. Although the clinical results are very controversial as the PRP preparation is not standardized between the studies. According to this, we searched the literature from 2020 to 2022 for clinical trials applying PRP and selected the 50 most relevant ones, which were scrutinized and the various features of the described protocols were compiled. Furthermore, we collected 10 whole blood samples that were split and processed into three protocols: PRP mild (≤1,000g), PRP high (>1,000g) and autologous platelet lysate (PLA). Also, we prepared platelet lysate heterologous (PLH) from 10 blood from donors. The protocols were evaluated for platelet counting, recovery and activity, Factor VIII activity and detection of a 40 biological factors. The platelet number and recovery were significantly higher (p <0,001) in the high protocol (72.03 ± 4.62 %) when compared to the mild one (44.58 ± 2.14 %). However, no significant difference was observed on platelet activity and Factor VIII comparing the protocols. PRP preparation protocols also did not interfere on expression pattern, where the main differences observed where in the comparisons of pooled and individual samples. Samples from the same donor were grouped together in spite of the preparation protocol. The PRP processing method did not affect the platelet physiological activity but can have effect on the biological factors availability and concentration. It can justify the choice of PRP processing depending on the pretended clinical application.