title: Osteoblast differentiation is enhanced by a nano-to-micro hybrid titanium surface created by Yb:YAG laser irradiation.
abstract: OBJECTIVE
  The aim of this study was to analyze the capacity of a new modified laser surface to stimulate calvarial osteoblasts isolated from neonatal mouse bones to differentiate and form mineralized nodules.
  
  
  METHODS
  Titanium discs were subjectezd or not to laser irradiation according to specific parameters and characterized. Osteoblasts isolated from neonatal mouse calvaria were cultured over the discs, and the capacity of these cells to proliferate (MTT assay), form mineralized nodules (Alizarin red assay), and enhance alkaline phosphatase activity (ALPase activity) was analyzed. Real-time PCR was used for quantification of gene expression.
  
  
  RESULTS
  Laser-irradiated titanium discs (L) presented a rough nano-to-micrometric oxidized surface contrasting with the smooth pattern on polished discs (P). The Ra on the micrometric level increased from 0.32 ± 0.01 μm on P surfaces to 10.57 ± 0.39 μm on L surfaces. When compared with P, L promoted changes in osteoblast morphology, increased mineralized nodule formation in osteoblasts cultured on the surfaces for 14 days, and enhanced ALPase activity at days 7 and 14. Transcription factors triggering osteoblast differentiation (Runx2 and Sp7) and genes encoding the bone extracellular matrix proteins collagen type-1 (Col1a1), osteopontin (Spp1), and osteocalcin (Bglap) were upregulated in cells on L surfaces compared with those on P surfaces at days 1-14.
  
  
  CONCLUSIONS
  Laser treatment of titanium surfaces created a rough surface that stimulated osteoblast differentiation.
  
  
  CONCLUSIONS
  Laser treatment of titanium generates a reproducible and efficient surface triggering osteoblast differentiation that can be of importance for osteointegration.
authors.affiliation: Department of Physiology and Pathology, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Mariscal-Muñoz
authors.affiliation: Department of Physiology and Pathology, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil.
authors.forename: Carlos A S
authors.initials: CA
authors.lastname: Costa
authors.affiliation: Department of Physiology and Pathology, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil.
authors.forename: Hewerson S
authors.initials: HS
authors.lastname: Tavares
authors.affiliation: Department of Physiology and Pathology, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil.
authors.forename: Jonas
authors.initials: J
authors.lastname: Bianchi
authors.affiliation: Department of Orthodontics and Pediatric Dentistry, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil.
authors.forename: Josimeri
authors.initials: J
authors.lastname: Hebling
authors.affiliation: National Institute for Space Research-INPE, São José dos Campos, São Paulo, 12227-010, Brazil.
authors.forename: João P B
authors.initials: JP
authors.lastname: Machado
authors.affiliation: Umeå University, Umeå, S-901 87, Sweden.|Sahlgrenska Academy at University of Gothenburg, Gothenburg, S-405 30, Sweden.
authors.forename: Ulf H
authors.initials: UH
authors.lastname: Lerner
authors.affiliation: Department of Physiology and Pathology, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil. souza.ppc@foar.unesp.br.
authors.forename: Pedro P C
authors.identifier: http://orcid.org/0000-0001-7266-6061
authors.initials: PP
authors.lastname: Souza
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D053844:Lasers, Solid-State; D051379:Mice; D008810:Mice, Inbred C57BL; D008855:Microscopy, Electron, Scanning; D016348:Osseointegration / Q000528:radiation effects; D010006:Osteoblasts / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D012886:Skull / Q000166:cytology*; D013052:Spectrometry, X-Ray Emission; D013499:Surface Properties; D014025:Titanium / Q000737:chemistry*
chemical_list: D014025:Titanium
keywords: Bone; Laser; Osteoblast; Titanium implants