Agradecimentos: This work was supported by the Research Financial Agency FAPESP (Brazil) [grant number 2014/27200-2, 2014] and by the Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT/MEC) through national funds, and co-financed by FEDER, under the Partnership...

Agradecimentos: This work was supported by the Research Financial Agency FAPESP (Brazil) [grant number 2014/27200-2, 2014] and by the Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT/MEC) through national funds, and co-financed by FEDER, under the Partnership Agreement PT2020 for the project M-ERA-NET/0004/2015-PAIRED and scholarship (SFRH/BD/130555/2017) to Ana Rita Fernandes

Platelet-rich plasma (PRP) associated with high molecular weight hyaluronic acid (HA) has been clinically used for tissue regeneration in orthopedics. Despite the recognized beneficial clinical outcomes (e.g., early pain control, improvement of patients' functional limitation and longer-term...

Platelet-rich plasma (PRP) associated with high molecular weight hyaluronic acid (HA) has been clinically used for tissue regeneration in orthopedics. Despite the recognized beneficial clinical outcomes (e.g., early pain control, improvement of patients' functional limitation and longer-term effectiveness compared to PRP and HA alone in mild and moderate osteoarthritis treatments), its use is still challenging and controversial due to lack of standardization of association practical protocols. Moreover, most studies neglect the matrix structure, that generates the ultimate properties of the association among platelets, fibrin network and the microparticles. In the present work, we aimed to analyze the influence of the PRP/HA association with a controlled matrix structure on the stability, rheological behavior, release of growth factors and in vitro proliferation of human adipose-derived mesenchymal cells (h-AdMSCs). The attenuation of the negative charge of HA was also evaluated. Pure PRP (P-PRP) (i.e., plasma enriched with platelets and poor in leukocytes) was prepared by centrifugation and activated with serum and calcium chloride (P-A-PRP). Autocrosslinked hyaluronic acid (AHA) was prepared by organocatalyzed auto-esterification and structured in microparticles ((MP)AHA) by shearing. The attenuation of the negative charge of (MP)AHA was performed with chitosan (CHT) by polyelectrolyte complexation yielding (MP)AHA-CHT. The results showed that microparticles (MPs) have viscoelastic properties, extrusion force and swelling ratio appropriate for injectable applications. The association of P-A-PRP with the controlled structure of (MP)AHA and (MP)AHA-CHT formed a matrix composed of platelets and of a fibrin network with fibers around 160 nm located preferably on the surface of the MPs with an average diameter of 250 mu m. Moreover, P-A-PRP/(MP)AHA and P-A-PRP/(MP)AHA-CHT associations were non-toxic and supported controlled growth factor (PDGF-AB and TGF-beta 1) release and in vitro proliferation of h-AdMSC with a similar pattern to that of P-A-PRP alone. The best h-AdMSC proliferation was obtained with the P-A-PRP/(MP)AHA-CHT75:25 indicating that the charge attenuation improved the cell proliferation. Thus, the association of P-A-PRP with the controlled structure of HA can be a valuable approach for orthopedic applications

FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP

2014/27200-2

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