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Open AccessArticle

Association of Platelet-Rich Plasma and Auto-Crosslinked Hyaluronic Acid Microparticles: Approach for Orthopedic Application

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Department of Engineering of Materials and Bioprocesses, School of Chemical Engineering, University of Campinas, 13083-852 Campinas, SP, Brazil
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Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra (FFUC), Pólo das Ciências da Saúde, 3000-548 Coimbra, Portugal
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Haematology and Hemotherapy Center, Umbilical Cord Blood Bank, University of Campinas, 13083-878 Campinas, SP, Brazil
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Institute of Bone and Cartilage (IOC), 13334-170 Indaiatuba, SP, Brazil
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Department of Pharmacy, Pharmaceutical Technology and Physical Chemistry, Faculty of pharmacy and Food Sciences, University of Barcelona, Institute of Nanoscience and Nanotechnology (IN2UB), 08028 Barcelona, Spain
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Centro de Investigación biomédica en red de enfermedades neurodegenerativas (CIBERNED), 08028 Barcelona, Spain
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CEB–Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal
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Authors to whom correspondence should be addressed.
Polymers 2019, 11(10), 1568; https://doi.org/10.3390/polym11101568
Received: 29 July 2019 / Revised: 25 September 2019 / Accepted: 25 September 2019 / Published: 26 September 2019
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 (AP-PRP). Autocrosslinked hyaluronic acid (AHA) was prepared by organocatalyzed auto-esterification and structured in microparticles (MPAHA) by shearing. The attenuation of the negative charge of MPAHA was performed with chitosan (CHT) by polyelectrolyte complexation yielding MPAHA-CHT. The results showed that microparticles (MPs) have viscoelastic properties, extrusion force and swelling ratio appropriate for injectable applications. The association of AP-PRP with the controlled structure of MPAHA and MPAHA-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 μm. Moreover, AP-PRP/MPAHA and AP-PRP/MPAHA-CHT associations were non-toxic and supported controlled growth factor (PDGF-AB and TGF-β1) release and in vitro proliferation of h-AdMSC with a similar pattern to that of AP-PRP alone. The best h-AdMSC proliferation was obtained with the AP-PRP/MPAHA-CHT75:25 indicating that the charge attenuation improved the cell proliferation. Thus, the association of AP-PRP with the controlled structure of HA can be a valuable approach for orthopedic applications. View Full-Text
Keywords: platelet-rich plasma; polyelectrolyte complexes; auto-crosslinked hyaluronic acid; chitosan; cell proliferation platelet-rich plasma; polyelectrolyte complexes; auto-crosslinked hyaluronic acid; chitosan; cell proliferation
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MDPI and ACS Style

Martins Shimojo, A.A.; Santos Duarte, A.S.; Santos Duarte Lana, J.F.; Malheiros Luzo, Â.C.; Fernandes, A.R.; Sanchez-Lopez, E.; Barbosa Souto, E.; Andrade Santana, M.H. Association of Platelet-Rich Plasma and Auto-Crosslinked Hyaluronic Acid Microparticles: Approach for Orthopedic Application. Polymers 2019, 11, 1568.

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