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Osteoblast differentiation is enhanced by a nano-to-micro hybrid titanium surface created by Yb:YAG laser irradiation

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Abstract

Objectives

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.

Conclusion

Laser treatment of titanium surfaces created a rough surface that stimulated osteoblast differentiation.

Clinical relevance

Laser treatment of titanium generates a reproducible and efficient surface triggering osteoblast differentiation that can be of importance for osteointegration.

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Acknowledgments

This research was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo-FAPESP (grants 2011/16351-1 and 2012/14912-9). The authors thank the Advanced Microscopy Laboratory from the Chemistry Institute of Araraquara for FEG-SEM facilities and Prof. Antonio Carlos Guastaldi for use of the laser facility. The authors also thank Antonio Fasano for his kind assistance on laser parameter characterization.

Conflict of interests

Authors declare not to have any influences that may potentially undermine the objectivity or integrity of the research.

Author information

Authors and Affiliations

  1. Department of Physiology and Pathology, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil

    Eduardo Mariscal-Muñoz, Carlos A. S. Costa, Hewerson S. Tavares, Jonas Bianchi & Pedro P. C. Souza

  2. Department of Orthodontics and Pediatric Dentistry, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil

    Josimeri Hebling

  3. National Institute for Space Research–INPE, São José dos Campos, São Paulo, 12227-010, Brazil

    João P.B. Machado

  4. Umeå University, Umeå, S-901 87, Sweden

    Ulf H. Lerner

  5. Sahlgrenska Academy at University of Gothenburg, Gothenburg, S-405 30, Sweden

    Ulf H. Lerner

Authors
  1. Eduardo Mariscal-Muñoz
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  2. Carlos A. S. Costa
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  3. Hewerson S. Tavares
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  4. Jonas Bianchi
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  7. Ulf H. Lerner
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  8. Pedro P. C. Souza
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Corresponding author

Correspondence to Pedro P. C. Souza.

Electronic supplementary material

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Supplemental 1

EDS analysis of P (a) and L (b) surfaces. (DOCX 265 kb)

Supplemental 2

XRD patterns for: (a) P sample; and (b) L sample. (DOC 543 kb)

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Mariscal-Muñoz, E., Costa, C.A.S., Tavares, H.S. et al. Osteoblast differentiation is enhanced by a nano-to-micro hybrid titanium surface created by Yb:YAG laser irradiation. Clin Oral Invest 20, 503–511 (2016). https://doi.org/10.1007/s00784-015-1533-1

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  • DOI: https://doi.org/10.1007/s00784-015-1533-1

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