Progress in Application of Concentrated Growth Factor in Oral Tissue Regeneration / 中国医学科学院学报

Tissue regeneration is an important engineering method for the treatment of oral soft and hard tissue defects.Growth factors,as one of the three elements of tissue regeneration,are a necessary condition for tissue regeneration.Concentrated growth factor(CGF)is a new generation of blood extract prepared by changing the centrifugal speed on the basis of the preparation of platelet-rich plasma(PRP)and platelet-rich fibrin(PRF).It contains abundant growth factors and a fibrin matrix with a three-dimensional network structure,being capable of activating angiogenesis and promoting tissue regeneration and healing.CGF has been widely used in the repair and regeneration of oral soft and hard tissues.This paper introduces the preparation and composition of CGF and reviews the application of CGF in oral implantation and the regeneration of oral bone tissue,periodontal tissue,and dental pulp tissue.

Platelet-rich Plasma Induces M2 Macrophage Polarization via Regulating AMPK Singling Pathway / 中国实验血液学杂志

OBJECTIVE@#To investigate the role of platelet-rich plasma (PRP) in inducing the M2 macrophage polarization via regulating AMPK singling pathway.@*METHODS@#The expressions of M1 marker CD11c and M2 marker CD206 in macrophages of blank control group, LPS group, LPS+PRP group, and LPS+PRP+Compound C group were detected by flow cytometry. Western blot was used to observe the effects of PRP on the expression of AMPK-mTOR signaling pathway-related proteins at different times (12 h, 18 h and 24 h) after LPS treatment. RNA interference technology was used to silence the expression of AMPK in macrophages, and the expression of TGF-β protein was subsequently examined by Western blot.@*RESULTS@#LPS significantly reduced the expression of CD206 and increased the expression of CD11c (P <0.05). After the addition of PRP, the expression of CD206 was significantly increased (P <0.05), while the expression of CD11c was significantly decreased (P <0.05). Compared with LPS group, PRP treatment significantly increased the expressions of p-AMPK and p-ULK1 proteins at 12 h, 18 h and 24 h, while significantly decreased the expression of p-mTOR protein (P <0.05). After the addition of AMPK inhibitor Compound C, the expression of CD206 was significantly reduced (P <0.05) and the expression of CD11c was significantly increased compared with LPS+PRP group (P <0.05). After silencing the expression of AMPK in macrophages, the promotion effect of PRP on TGF-β was significantly reduced (P <0.05).@*CONCLUSION@#PRP can stimulate the transformation of macrophages to M2 type via AMPK signalling pathway.

Cost and effectiveness of Platelet Rich Plasma in the healing of varicose ulcer: Meta-analysis / Costo y efectividad del Plasma Rico en Plaquetas en la cicatrización de úlcera varicosa: Metaanálisis / Custo e efetividade do Plasma Rico em Plaquetas na cicatrização de úlcera varicosa: Metanálise

ABSTRACT Objectives: to analyze the evidence on the cost and effectiveness of Plaque Rich Plasma in the treatment of venous ulcers compared to other topical therapies. Methods: systematic review, with search in the databases: COCHRANE, EMBASE, MEDLINE via PubMed, LILACS, CINAHL, SCOPUS, without temporal cut and in the English, Portuguese and Spanish languages. Results: fifteen articles were included, a cost-minimization analysis showed that the cost of Plaque Rich Plasma is € 163.00 ± 65.90, slightly higher than the cost of standard dressing. Regarding effectiveness, the results of the studies associated with the meta-analysis suggest a tendency that Plaque Rich Plasma is effective in the healing of venous ulcers. Conclusions: it is concluded that there are few studies about the cost of Platelet Rich Plasma and this product tends to be effective in the healing of venous ulcers. However, more controlled and randomized clinical studies are necessary in order to establish a stronger recommendation.

RESUMEN Objetivos: analizar las evidencias acerca del costo y de la efectividad del Plasma Rico en Plaquetas en el tratamiento de úlceras venosas comparado a las otras terapias tópicas. Métodos: revisión sistemática con la búsqueda en bases de datos: Cochrane Library, EMBASE, MEDLINE vía PubMed, LILACS, CINAHL, SCOPUS, hay un tiempo y en inglés, portugués y español. Resultados: se incluyeron 15 artículos, un análisis de costo-minimización demostró que el costo del Plasma Rico en Plaquetas es de € 163,00 ± 65,90, poco superior al costo del vendaje estándar. En cuanto a la efectividad, los resultados de los estudios asociados al metaanálisis sugieren una tendencia de que el Plasma Rico en Plaquetas es efectivo en la cicatrización de las úlceras venosas. Conclusiones: se concluye que hay pocos estudios acerca del costo del Plasma Rico en Plaquetas y que ese tiende a ser efectivo en la cicatrización de úlceras venosas. Todavía, son necesarios más estudios clínicos controlados y aleatorizados para que se pueda establecer una recomendación más fuerte.

RESUMO Objetivos: analisar as evidências acerca do custo e da efetividade do Plasma Rico em Plaquetas no tratamento de úlceras venosas comparado às outras terapias tópicas. Métodos: revisão sistemática, com busca nas bases de dados COCHRANE, EMBASE, MEDLINE via PubMed, LILACS, CINAHL, SCOPUS, sem recorte temporal e nos idiomas inglês, português e espanhol. Resultados: foram incluídos 15 artigos. Uma análise de custo-minimização demonstrou que o custo do Plasma Rico em Plaquetas é de €163,00 ± 65,90, pouco superior ao custo do curativo padrão. Quanto à efetividade, os resultados dos estudos associados à metanálise sugerem uma tendência de que o Plasma Rico em Plaquetas é efetivo na cicatrização das úlceras venosas. Conclusões: conclui-se que há poucos estudos acerca do custo do Plasma Rico em Plaquetas e esse produto tende a ser efetivo na cicatrização de úlceras venosas. Entretanto, são necessários mais estudos clínicos controlados e randomizados para que se possa estabelecer uma recomendação mais forte.

Aplicación tópica de plaquetas plasmáticas en pacientes con queratitis punteada superficial / Topical use of plasmatic platelets in patients with superficial punctuated queratitis

Se realizó un estudio clínico, prospectivo, intervencionista y comparativo, de 41 pacientes (31 ojos) con diagnóstico de queratitis punteada superficial -seleccionados por muestreo aleatorio simple-, atendidos en el Hospital Infantil Docente Sur Dr Antonio María Béguez César de Santiago de Cuba, desde noviembre del 2016 hasta mayo del 2017, con vistas a demostrar la eficacia del tratamiento tópico con colirio de plaquetas plasmáticas en ellos, para lo cual se conformaron 2 grupos: el A, que recibió tratamiento tópico convencional, y el B con la misma terapia más colirio de plasma rico en plaquetas. Los pacientes tuvieron seguimiento clínico a las dos y cuatro semanas de aplicado el tratamiento. En la mayoría de los pacientes del grupo B el tiempo de cicatrización corneal fue de 7 días y el de curación de 2 semanas; de manera que se demostró la eficacia de este colirio autólogo en la cicatrización corneal, sin riesgo de reacción inmunológica y de bajo costo económico.

A clinical, prospective, interventionist and comparative study of 41 patients (31 eyes) with diagnosis of superficial punctate queratitis -selected by simple random sampling-, assisted in Dr Antonio María Béguez Caesar Southern Teaching Pediatric Hospital in Santiago de Cuba was carried out from November, 2016 to May, 2017, with the objective of demonstrating the effectiveness of the topical treatment of plasmatic platelets in them with eyewash, for which 2 groups were formed: group A with conventional topical treatment, and group B with the same therapy plus eyewash with rich platelets plasm. The patients had clinical follow- up at two and four weeks of having applied the treatment. In most of the patients of group B the time of corneal scaring was of 7 days and that of cure of 2 weeks; so that the effectiveness of this autologous eyewash was demonstrated in the corneal scaring, without risk of immunological reaction and with low economic cost.

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

Canine mesenchymal cells (MSCs) derived from Wharton's jelly were co-cultured, then supplemented or not supplemented with platelet rich plasma (PRP) and demineralized bone matrix (DBM) to verify osteogenic differentiation. Osteoblastic differentiation followed by mineralized bone matrix production was found to be significantly higher (p < 0.05) when MSCs were associated with PRP/DBM in culture after 14-21-days of induction. Osteopontin and osteocalcin gene expression were significantly superior (p < 0.05) under the same culture conditions after 21 days of observation. In conclusion, addition of PRP to DBM co-cultured with MSCs successfully induced osteogenesis in vitro.

Avaliação da eficácia de diferentes protocolos de preparo do Plasma Rico em Plaquetas para uso em Medicina Equina / Evaluating the effectiveness of different protocols for preparation of platelet rich plasma for use in equine medicine

O Plasma Rico em Plaquetas (PRP) é um preparado do sangue total que contém diversos fatores de crescimento responsáveis pela proliferação e diferenciação celular, angiogênese, como também pelo aumento da produção da matriz extracelular. Nesse sentido, o objetivo do presente estudo foi testar 10 protocolos diferentes de centrifugação para obtenção de PRP a partir do sangue total de equinos hígidos. Para isso foram utilizadas 10 amostras de 27mL de sangue total de cinco animais, as quais foram centrifugadas conforme cada protocolo proposto. Os resultados revelaram que os protocolos com menor força de centrifugação relativa resultaram em maior (p<0,05) concentração de plaquetas e, que não houve (p>0,05) influência do tempo de centrifugação em relação a essa variável. A influência do tempo foi observada apenas no número de leucócitos em protocolos com menor força de centrifugação relativa (FCR). Os quatro melhores protocolos, que obtiveram as maiores concentrações de plaquetas, foram submetidos à análise pelo teste de ELISA para dosar a quantidade de TGF-β que não revelou diferença (p>0,05) entre os protocolos.

Platelet-rich plasma (PRP) is a prepared of the whole blood which contains several growth factors responsible for cellular proliferation and differentiation, angiogenesis, and for the increase of the extracellular matrix. Thus, the aim of this study was to test 10 different centrifugation protocols to obtain PRP from the whole blood of healthy equines. Ten samples of 27mL of the whole blood of 5 healthy equines were used, which were centrifuged in accordance to the proposed protocols. The results showed that the protocols with less relative centrifugation force resulted in greater (p<0,05) platelets concentration. Also, platelets concentration was not influenced by varying the time of centrifugation. However, time did affect the number of leukocytes in some protocols. The best four protocols were analyzed by ELISA test to quantitate the amount of TGF-β, which revealed no difference (p> 0.05) between the protocols.

Platelet-rich plasma inhibits the apoptosis of highly adipogenic homogeneous preadipocytes in an in vitro culture system

Auto-transplantation of adipose tissue is commonly used for the treatment of tissue defects in plastic surgery. The survival of the transplanted adipose tissue is not always constant, and one of reasons is the accelerated apoptosis of the implanted preadipocytes. We have recently established highly homogeneous preadipocytes, named ccdPAs. The aim of the current study was to evaluate the regulation of the potency of platelet-rich plasma (PRP) on the apoptosis of ccdPAs in vitro. PRP stimulated the proliferation of the preadipocytes in a dose-dependent manner, and the stimulatory activity of 2% PRP was significantly higher than that of 2% FBS or 2% platelet-poor plasma (PPP). The presence of 2% PRP significantly inhibited serum starvation- or TNF-alpha/cycloheximide-induced apoptosis in comparison to 2% FBS or 2% PPP. DAPK1 and Bcl-2-interacting mediator of cell death (BIM) mRNAs were reduced in the preadipocytes cultured with 2% PRP in comparison to those cultured in 2% FBS. The gene expression levels were significantly higher in cells cultured without serum in comparison to cells cultured with 2% FBS, and the levels in the cells with 2% PRP were reduced to 5-10% of those in the cells without serum. These results indicated that ccdPAs exhibit anti-apoptotic activities, in addition to increased proliferation, when cultured in 2% PRP in comparison to the same concentration of FBS, and that this was accompanied with reduced levels of DAPK1 and BIM mRNA expression in in vitro culture. PRP may improve the outcome of transplantation of adipose tissue by enhancing the anti-apoptotic activities of the implanted preadipocytes.

Enhanced skin wound healing by a sustained release of growth factors contained in platelet-rich plasma

Platelet-rich plasma (PRP) contains growth factors that promote tissue regeneration. Previously, we showed that heparin-conjugated fibrin (HCF) exerts the sustained release of growth factors with affinity for heparin. Here, we hypothesize that treatment of skin wound with a mixture of PRP and HCF exerts sustained release of several growth factors contained in PRP and promotes skin wound healing. The release of fibroblast growth factor 2, platelet-derived growth factor-BB, and vascular endothelial growth factor contained in PRP from HCF was sustained for a longer period than those from PRP, calcium-activated PRP (C-PRP), or a mixture of fibrin and PRP (F-PRP). Treatment of full-thickness skin wounds in mice with HCF-PRP resulted in much faster wound closure as well as dermal and epidermal regeneration at day 12 compared to treatment with either C-PRP or F-PRP. Enhanced skin regeneration observed in HCF-PRP group may have been at least partially due to enhanced angiogenesis in the wound beds. Therefore, this method could be useful for skin wound treatment.

Individual Variation in Growth Factor Concentrations in Platelet-rich Plasma and Its Influence on Human Mesenchymal Stem Cells / 대한진단검사의학회지

BACKGROUND: The objective of this study was to explore whether individual variations in the concentration of growth factors (GFs) influence the biologic effects of platelet-rich plasma (PRP) on human mesenchymal stem cells (HMSCs). METHODS: The concentrations of 7 representative GFs in activated PRP (aPRP) were measured using ELISA. The effects of PRP on the proliferation and alkaline phosphatase (ALP) activity of HMSCs were examined using several concentrations of aPRP from 3 donors; the relationships between the GF levels and these biologic effects were then evaluated using 10% aPRP from 5 subgroups derived from 39 total donors. HMSCs were cultured in DMEM with the addition of aPRP for 4 or 12 days; then, DNA content and ALP activity were measured. RESULTS: The quantity of DNA increased significantly at a 10% concentration of aPRP, but the ALP activity was suppressed at this concentration of aPRP. The GF concentrations varied among donors, and 5 subgroups of characteristic GF release patterns were identified via cluster analysis. DNA levels differed significantly between groups and tended to be higher in groups with higher concentrations of transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factors (PDGFs). DNA quantity was positively correlated with TGF-beta1 concentration, and was negatively correlated with donor age. ALP activity was negatively correlated with PDGF-BB concentration. CONCLUSIONS: The varying GF concentrations may result in different biologic effects; thus, individual differences in GF levels should be considered for reliable interpretation of the biologic functions and standardized application of PRP.

Platelet concentration of plateletrich plasma from dogs, obtained through three centrifugation speeds

The platelets release at least 4 growth factors (Platelet Derived Growth Factor. â1 and â2 Transforming Growth Factors and Insulin-like Growth Factor) which are responsible for the migration and activation of cells that will start the reparation of soft tissues and bones. The Platelet Rich Plasma is an autogenous source for Growth Factors, obtained by platelet concentration by centrifuging total blood. This study aimed the comparison of platelet concentrations in plasma centrifuged in three different centrifugation speeds (1300, 1600 e 3200rpm), for the production of platelet rich plasma. Blood was drowned from 15 dogs, 40ml of each, and these were divided into four groups and centrifuged at 800rpm. Then the first group was centrifuged at 1300rpm, the second at 1600rpm, the third at 3200rpm and the last was used as control, named plasma. The mean percentage increase in the platelet concentration for each technique was: 1300 –183%, 1600 – 210% and 3200 – 222%. But in centrifugation at 3200rpm, platelets presented altered morphology and different sizes in every sample studied, which was understood as severe cell damage. It was concluded that the best technique for the preparation of the platelet rich plasma in dogs consisted of the previous centrifugation of the blood at 800rpm for ten minutes, and then the plasma should be separated. This plasma is then submitted to a second centrifugation of 1600rpm for 10 minutes, and the platelet poor plasma is separated and discharged.

Plaquetas liberam ao menos quatro fatores de crescimento ( Fator de Crescimento derivado de Plaquetas, Fatores de transformação de crescimento â1 and â2 e Fator de crescimento semelhante a insulina) responsáveis pela migração e ativação de células que iniciarão os processos de reparação de tecidos moles e ossos. O Plasma Rico em Plaquetas é fonte autógena de fatores de crescimento, obtida pela concentração das plaquetas através de centrifugação de sangue total. Este estudo visa a comparação das concentrações plaquetárias no plasma obtidas por três diferentes velocidades de centrifugação (1300,1600 e 3200 rpm), para produção de Plasma Rico em Plaquetas. 40 ml de sangue total foram retirados de cada animal, divididos em quatro grupos, e centrifugados inicialmente a 800 rpm. A seguir, as amostras do primeiro grupo foram centrifugadas a 1300 rpm, as do Segundo a 1600 rpm, as do terceiro a 3200 rpm e as do quarto grupo foram usadas como controle, denominadas plasma. O aumento médio da porcentagem na concentração de plaquetas para cada técnica foi: 1300- 183%, 1600 - 210% e 3200 - 222%. No entanto, a centrifugação a 3200 rpm, as plaquetas apresentaram a morfologia alterada e tamanhos diferentes em cada amostra estudada, que foram compreendidas como danos celulares severos. Como conclusão deste estudo, obteve-se que a melhor técnica para a preparação do plasma rico em plaquetas de cães consiste na centrifugação precedente do sangue em 800 rpm por dez minutos, separando o plasma, sendo este submetido a uma segunda centrifugação de 1600 rpm por 10 minutos, separando e desprezando o plasma pobre em plaquetas.