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Treatment With a Growth Factor–Protein Mixture Inhibits Formation of Mineralized Nodules in Osteogenic Cell Cultures Grown on Titanium

Marcos Andrade de Oliva, William Marcatti Amarú Maximiano, Larissa Moreira Spínola de Castro, Paulo Eliandro da Silva, Jr, Roger Rodrigo Fernandes, Pietro Ciancaglini, Márcio Mateus Beloti, Antonio Nanci, Adalberto Luiz Rosa and Paulo Tambasco de Oliveira

Cell Culture Laboratory, School of Dentistry of Ribeirão Preto (MAO,WMAM,LMSC,PES,RRF,MMB,ALR,PTO) and Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (PC), Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil; and Laboratory for the Study of Calcified Tissues and Biomaterials, Université de Montréal, Montréal, Quebec, Canada (AN)

Correspondence to: Prof. Dr. Paulo Tambasco de Oliveira, Division of Oral Histology, School of Dentistry of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-904 Ribeirão Preto, SP, Brazil. E-mail: tambasco{at}usp.br

Despite wide clinical application, the efficacy of platelet-rich plasma (PRP) for repairing bone defects and enhancing osseointegration of metal implants is still subject of debate. This study aimed to evaluate the effects of a well-defined PRP-like mixture containing platelet-derived growth factor-BB, transforming growth factor (TGF)-β1, TGF-β2, albumin, fibronectin, and thrombospondin [growth factors (GFs) + proteins] on the development of the osteogenic phenotype on titanium (Ti) in vitro. Human alveolar bone-derived osteoblastic cells were subcultured on Ti discs and exposed during the first 7 days to osteogenic medium supplemented with GFs + proteins and to osteogenic medium alone thereafter up to 14 days. Control cultures were exposed to only osteogenic medium. Dose–response experiments were carried out using rat primary calvarial cells exposed to GFs + proteins and 1:10 or 1:100 dilutions of the mixture. Treated human-derived cell cultures exhibited a significantly higher number of cycling cells at days 1 and 4 and of total cells at days 4 and 7, significantly reduced alkaline phosphatase (ALP) activity at days 4, 7, and 10, and no Alizarin red–stained areas (calcium deposits) at day 14, indicating an impairment in osteoblast differentiation. Although the 1:10 and 1:100 dilutions of the mixture restored the proliferative activity of rat-derived osteogenic cells to control levels and promoted a significant increase in ALP activity at day 10 compared with GFs + proteins, mineralized nodule formation was only observed with the 1:100 dilution (50% of the control). These results showed that a PRP-like protein mixture inhibits development of the osteogenic phenotype in both human and rat osteoblastic cell cultures grown on Ti. (J Histochem Cytochem 57:265–276, 2009)

Key Words: cell culture • osteoblasts • growth factors • cell proliferation • cell differentiation • mineralization • titanium

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