Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 9, 1741-55
eCollection

Osteogenic Activity of Titanium Surfaces With Nanonetwork Structures

Affiliations

Osteogenic Activity of Titanium Surfaces With Nanonetwork Structures

Helin Xing et al. Int J Nanomedicine.

Abstract

Background: Titanium surfaces play an important role in affecting osseointegration of dental implants. Previous studies have shown that the titania nanotube promotes osseointegration by enhancing osteogenic differentiation. Only relatively recently have the effects of titanium surfaces with other nanostructures on osteogenic differentiation been investigated.

Methods: In this study, we used NaOH solutions with concentrations of 2.5, 5.0, 7.5, 10.0, and 12.5 M to develop a simple and useful titanium surface modification that introduces the nanonetwork structures with titania nanosheet (TNS) nanofeatures to the surface of titanium disks. The effects of such a modified nanonetwork structure, with different alkaline concentrations on the osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMMSCs), were evaluated.

Results: The nanonetwork structures with TNS nanofeatures induced by alkali etching markedly enhanced BMMSC functions of cell adhesion and osteogenesis-related gene expression, and other cell behaviors such as proliferation, alkaline phosphatase activity, extracellular matrix deposition, and mineralization were also significantly increased. These effects were most pronounced when the concentration of NaOH was 10.0 M.

Conclusion: The results suggest that nanonetwork structures with TNS nanofeatures improved BMMSC proliferation and induced BMMSC osteogenic differentiation. In addition, the surfaces formed with 10.0 M NaOH suggest the potential to improve the clinical performance of dental implants.

Keywords: bone marrow mesenchymal stem cells; nanotopography; osseointegration; surface modification.

Figures

Figure 1
Figure 1
Scanning electron micrographs after treatment with various concentrations of NaOH and AFM of specimen surfaces. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Lower magnification of ×10,000, showing the overall microscale topography (upper images). Higher magnification of ×50,000, reveals the nanoscale texture (middle images). AFM of specimen surfaces, showing three-dimensional images and roughness of CON and experimental groups (lower images). Abbreviations: CON, control titanium surface; AFM, atomic force microscopy.
Figure 2
Figure 2
Contact angle measurements of ultrapure water droplets pipetted on specimens. (A) Optical images. (B) Quantitative degree results. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Statistical significance: a, P<0.01 vs 2.5 M; b, P<0.01 vs 5.0 M; c, P<0.01 vs 7.5 M; d, P<0.01 vs 10.0 M; e, P<0.01 vs 12.5 M. Abbreviation: CON, control titanium surface.
Figure 3
Figure 3
Assay of protein adsorption to different specimens after 1, 3, 6, and 24 hours incubation in bovine serum albumin. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Statistical significance: a, P<0.01 vs CON; b, P<0.01 vs 2.5 M; c, P<0.01 vs 5.0 M; d, P<0.01 vs 7.5 M; e, P<0.01 vs 12.5 M. Abbreviation: CON, control titanium surface.
Figure 4
Figure 4
Initial number of adherent BMMSCs, measured by counting cells stained with DAPI under a fluorescence microscope after 30 minutes, 1 hour, and 3 hours incubation. (A) Fluorescence images of cells attached after 3 hours incubation. (B) Quantitative results of initial number of adherent BMMSCs. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Statistical significance: a, P<0.01 vs CON; b, P<0.01 vs 2.5 M; c, P<0.01 vs 5.0 M; d, P<0.01 vs 7.5 M; e, P<0.01 vs 12.5 M. Abbreviations: CON, control titanium surface; BMMSCs, bone marrow mesenchymal stem cells; DAPI, 4′,6-diamidino-2-phenylindole.
Figure 5
Figure 5
Cell proliferation on samples after 1, 3, and 7 days of incubation, measured by the CellTiter-Blue® Cell Viability Assay. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Statistical significance: a, P<0.01 vs CON; b, P<0.01 vs 2.5 M; c, P<0.01 vs 5.0 M; d, P<0.01 vs 7.5 M; e, P<0.01 vs 12.5 M. CellTiter-Blue® Cell Viability Assay (Promega Corporation, Madison, WI, USA). Abbreviation: CON, control titanium surface.
Figure 6
Figure 6
Scanning electron micrographs of cells after 30 minutes (A), 1 hour (B), 3 hours (C), and 3 days (D) culture. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Pictures at lower magnification (×5,000) show the morphology of single cells. Pictures at higher magnification (×20,000) show the detailed interaction of the cell with the nanostructure. Abbreviations: CON, control titanium surface; BMMSCs, bone marrow mesenchymal stem cells.
Figure 7
Figure 7
Alkaline phosphatase staining of mesenchymal stem cells after 1 week (A) and 2 weeks (B) culture. Alkaline phosphatase activity of mesenchymal stem cells on different samples cultured for 1 and 2 weeks (C). Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Statistical significance: a, P<0.01 vs CON; b, P<0.01 vs 2.5 M; c, P<0.01 vs 5.0 M; d, P<0.01 vs 7.5 M; e, P<0.01 vs 12.5 M. Abbreviations: CON, control titanium surface; ALP, alkaline phosphatase.
Figure 8
Figure 8
Extracellular matrix mineralization on different samples after 3 weeks (A) and 4 weeks (B) culture of mesenchymal stem cells. (C) Quantitative results of calcium deposition. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Statistical significance: a, P<0.05 vs CON; b, P<0.05 vs 2.5 M; c, P<0.05 vs 5.0 M; d, P<0.05 vs 7.5 M; e, P<0.05 vs 12.5 M. Abbreviation: CON, control titanium surface.
Figure 9
Figure 9
OCN production after 3 and 4 weeks of culture, as measured by sandwich enzyme immunoassay. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Statistical significance: a, P<0.01 vs CON; b, P<0.01 vs 2.5 M; c, P<0.01 vs 5.0 M; d, P<0.01 vs 7.5 M; e, P<0.01 vs 12.5 M. Abbreviations: CON, control titanium surface; OCN, osteocalcin.
Figure 10
Figure 10
Gene expression in primary osteoblasts cultured on titanium surfaces after incubation for 3 and 7 days: (A) BSP, (B) ON, (C) RUNX2, and (D) COL-1. Data were generated by real-time polymerase chain reaction and are shown as mean ± standard deviation expression relative to GADPH. Notes: 2.5, 5.0, 7.5, 10.0, and 12.5 M correspond to the concentration of NaOH. Statistical significance: a, P<0.01 vs CON; b, P<0.01 vs 2.5 M; c, P<0.01 vs 5.0 M; d, P<0.01 vs 7.5 M; e, P<0.01 vs 12.5 M. Abbreviations: CON, control titanium surface; OCN, osteocalcin; BSP, bone sialoprotein; ON, osteonectin; RUNX2, runt-related transcription factor 2; COL-1, collagen type 1; GADPH, glyceraldehyde-3-phosphate dehydrogenase; mRNA, messenger ribonucleic acid.

Similar articles

See all similar articles

Cited by 14 articles

See all "Cited by" articles

References

    1. Rosa MB, Albrektsson T, Francischone CE, Schwartz Filho HO, Wennerberg A. The influence of surface treatment on the implant roughness pattern. J Appl Oral Sci. 2012;20(5):550–555. - PMC - PubMed
    1. Meulen Pv, Linden Wv, Eeden Rv. Optimal restoration of dental esthetics and function with advanced implant-supported prostheses: a clinical report. J Prosthodont. 2012;21(5):393–399. - PubMed
    1. Nanci A, Wuest JD, Peru L, et al. Chemical modification of titanium surfaces for covalent attachment of biological molecules. J Biomed Mater Res. 1998;40(2):324–335. - PubMed
    1. Buser D, Broggini N, Wieland M, et al. Enhanced bone apposition to a chemically modified SLA titanium surface. J Dent Res. 2004;83(7):529–533. - PubMed
    1. Guo J, Padilla RJ, Ambrose W, De Kok IJ, Cooper LF. The effect of hydrofluoric acid treatment of TiO2 grit blasted titanium implants on adherent osteoblast gene expression in vitro and in vivo. Biomaterials. 2007;28(36):5418–5425. - PubMed

Publication types

Feedback