In vitro evaluation of osteoblast responses to carbon nanotube-coated titanium surfaces

Abstract

BackgroundThe effects of surface roughness and carboxyl functionalization of multi-walled carbon nanotubes (MWCNTs) mixed with collagen coated onto titanium (Ti) substrates on MC3T3-E1 osteoblasts were evaluated.MethodsThe proliferation, differentiation, and matrix mineralization were investigated using (1) smooth-surfaced Ti discs, (2) Ti discs coated with collagen and MWCNT (Ti-MWCNT), and (3) Ti discs coated with collagen and MWCNT-COOH (Ti-MWCNT-COOH) for applications in orthodontic mini screw implants (MSIs). The coatings were uniform when analyzed using scanning electron microscopy (SEM), and surface roughness was evaluated by surface profilometry that demonstrated similar surface roughness (Ra, mean ± SD) in the MWCNT (0.83 ± 0.02 μm) and MWCNT-COOH (0.84 ± 0.01 μm) groups. MTT (3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide) assay was performed after days 1, 3, and 7 to assess proliferation. Alkaline phosphatase (ALP)-specific activity was assessed after day 7 to quantify differentiation. Alizarin red staining was measured after day 28 to quantify matrix mineralization. All data were analyzed with JMP Pro11 software (SAS, USA) with a statistical significance of p < 0.05.ResultsSurface profilometry demonstrated similar surface roughness (Ra, mean ± SD) in the MWCNT (0.83 ± 0.02 μm) and MWCNT-COOH (0.84 ± 0.01 μm) groups. On day 7, ALP assay showed that MWCNT-COOH (mean ± SD 0.98 ± 0.26 U/μg of protein) enhanced cell differentiation when compared to the uncoated group (p = 0.05). Alizarin red staining after 28 days of cell culture revealed that MWCNT-COOH (mean ± SD 1.5 ± 0.2 OD405) increased (p = 0.03) matrix mineralization when compared to the uncoated group (0.9 ± 0.09 OD405).ConclusionsThis study showed that coatings containing MWCNT-COOH (increased hydrophilic surface chemistry) influence osteoblast proliferation, differentiation, and matrix mineralization and should be further studied for applications in orthodontic MSIs.

Enhanced growth and osteogenic differentiation of MC3T3-E1 cells on Ti6Al4V alloys modified with reduced graphene oxide

“…In order to detect and quantify calcium in the mineralized nodules, cells were treated with alizarin red staining (ARS) solution on day 14. 48 The cells cultured on different substrates for 14 days were washed with PBS, xed with 70% ethanol for 30 minutes, and stained with alizarin red solution for 30 minutes at 37 C. Then, the titanium substrates were washed with PBS three times to remove the non-specic staining. Subsequently, the mineralized nodule was dissolved with 10% cetylpyridinium chloride solution for 30 minutes with gentle shaking, and the absorbance at 562 nm was measured with a microplate reader 49 (Tecan innite M200, Tecan Inc., Switzerland).…”

Análise Do Percentual De Colágeno Na Dentina Humana Entre Diferentes Etnias, Gêneros E Idades

“…There are countless tests that evaluate cell viability, however, some stand out for being widely used. Among these testes, MTT (Aranha, 2014;Abreu, 2008;Gau, Lai, Leung, 2012;Junior, 2012;Wu et al, 2017;Baili et al, 2016;Galeotti et al, 2013;Hao et al, 2017;Kandiah et al, 2017;Karahalil et al, 2014;Preedy, Perni, Prokopovich, 2017;Silva, 2011;Subramani et al, 2016), MTS (Silva, 2011;Chellini et al, 2017, Gosau et al, 2016Lee et al, 2017), XTT (Aranha, 2014;Coelho, 2013, Fotakis e Timbrell, 2006, Neutral Red (Abreu, 2008;Azevedo, Cruz, Pinto, 2006;Pereira, 2008;Rogero et al, 2003), Alamar Blue (Aranha, 2014;Gao, Lai, Leung, 2012;Gosau et al, 2016;Alqhtani et al, 2017), and the synthesis of nitric oxide (Felgueiras, 2011;Parida et al, 2017;Silva, 2017;Yarlagadda et al, 2017;Zhang, Li, Yang, 2015) can be mentioned.…”

Contamination of titanium dental implants: a narrative review

“…Carboxylates can be found from the coating of Ti surfaces. Carboxylated multi-walled carbon nanotube coated Ti has been shown to have increased osteoblast proliferation, differentiation, and matrix mineralization [54]. However, carboxylates contaminants can cause adverse effects on Ti surface.…”