title: Treatment with a growth factor-protein mixture inhibits formation of mineralized nodules in osteogenic cell cultures grown on titanium.
abstract: 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)-beta1, TGF-beta2, 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 ( approximately 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.
authors.affiliation: Cell Culture Laboratory, School of Dentistry of Ribeirão Preto, Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (PC), Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Marcos Andrade
authors.initials: MA
authors.lastname: de Oliva
authors.forename: William Marcatti Amarú
authors.initials: WM
authors.lastname: Maximiano
authors.forename: Larissa Moreira Spínola
authors.initials: LM
authors.lastname: de Castro
authors.forename: Paulo Eliandro
authors.initials: PE
authors.lastname: da Silva
authors.forename: Roger Rodrigo
authors.initials: RR
authors.lastname: Fernandes
authors.forename: Pietro
authors.initials: P
authors.lastname: Ciancaglini
authors.forename: Márcio Mateus
authors.initials: MM
authors.lastname: Beloti
authors.forename: Antonio
authors.initials: A
authors.lastname: Nanci
authors.forename: Adalberto Luiz
authors.initials: AL
authors.lastname: Rosa
authors.forename: Paulo Tambasco
authors.initials: PT
authors.lastname: de Oliveira
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000166:cytology; D002113:Calcification, Physiologic; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005353:Fibronectins / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D008855:Microscopy, Electron, Scanning; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects*; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D053657:Platelet-Rich Plasma; D051381:Rats; D012709:Serum Albumin / Q000494:pharmacology*; D019699:Thrombospondins / Q000494:pharmacology*; D013997:Time Factors; D023822:Tissue Engineering; D014025:Titanium; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D053781:Transforming Growth Factor beta2 / Q000494:pharmacology
chemical_list: D005353:Fibronectins; D036341:Intercellular Signaling Peptides and Proteins; D010982:Platelet-Derived Growth Factor; D012709:Serum Albumin; D019699:Thrombospondins; D053773:Transforming Growth Factor beta1; D053781:Transforming Growth Factor beta2; D014025:Titanium