Design and in vitro evaluation of simvastatin-hydroxyapatite coatings by an electrochemical process on titanium surfaces.

Design and in vitro evaluation of simvastatin-hydroxyapatite coatings by an electrochemical process on titanium surfaces.

Journal of biomedical nanotechnology (2014-05-09)

Zhao Shifang, Shi Jue, He Fuming, Li Liu, Yang Guoli

ABSTRACT

The aim of this study was to deposit simvastatin-hydroxyapatite (sim-HA) coatings on titanium surfaces by an electrochemical process and evaluate osteoprogenitor cell responses to sim-HA-coated surfaces. Simvastatin was prepared onto titanium surfaces with varying concentration (10(-7), 10(-6), 10(-5) and 10(-4) mol/L). Surface characteristics were performed by FSEM, XRD and FTIR. LC-MS/MS method was used for simvastatin quantification in Sim-HA-coated surfaces. Murine calvaria-derived pre-osteoblastic cell (MC3T3-E1) proliferation, alkaline phosphatase activity (ALP) and osteocalcin release were used to measure osteoblastic activities. FSEM observation showed rod-like HA crystals covered on all surfaces. As drug concentration increased, the crystal diameter decreased. XRD and FTIR observations showed all coatings consisted of HA. LC-MS/MS test showed the simvastatin concentration in 10(-6) mol/L group was 2.77 x 10(-7) mol/L/cm2, while in 10(-7) mol/L group was 1.89 x 10(-7) mol/L/cm2. MC3T3-E1 cells grown on 10(-7) mol/L and 10(-6) mol/L Sim-HA surfaces showed increased ALP activity as compared to HA-coated surfaces on day 7 (P < 0.05) while at 10 day, all Sim-HA groups were significantly increased than HA group (P < 0.05). Two Sim-HA groups showed significant increases in osteocalcin production on 7 10 and 14 day (P < 0.05). It showed that incubation with 10(-7) mol/L simvastatin enhanced the cell proliferation to a statistically significant extent (P < 0.05) compared with the HA group at day 7. It was concluded that simvastatin was successfully deposited into HA coatings using the electrochemical process and the sim-HA coatings enhanced differentiation of osteoprogenitor cells.

MATERIALS

Product Number

Brand

Product Description

369489

Sigma-Aldrich

Titanium, foil, thickness 2.0 mm, 99.7% trace metals basis

460400

Sigma-Aldrich

Titanium, wire, diam. 0.25 mm, 99.7% trace metals basis

04238

Sigma-Aldrich

Hydroxyapatite, puriss., meets analytical specification of Ph. Eur., BP, FCC, E341, ≥90% (calculated on glowed substance)

P8361

Sigma-Aldrich

Phosphatase, Alkaline bovine, recombinant, expressed in Pichia pastoris, ≥4000 units/mg protein

D6898

Sigma-Aldrich

Deoxyribonucleic acid sodium salt from herring testes, Type XIV

D4154

Sigma-Aldrich

Plasmid DNA from E. coli RRI, pUC18, buffered aqueous solution

D3035

Sigma-Aldrich

Deoxyribonucleic acid from human placenta, buffered aqueous solution, sexed, female

D3404

Sigma-Aldrich

Plasmid DNA from E. coli RRI, pUC19, buffered aqueous solution

21223

Sigma-Aldrich

Hydroxyapatite, purum p.a., ≥90% (as Ca₃(PO₄)₂, KT)

GF23860144

Titanium, tube, 500mm, outside diameter 6.35mm, inside diameter 4.57mm, wall thickness 0.89mm, annealed, 99.6+%

GF12869062

Titanium, wire reel, 1m, diameter 0.25mm, as drawn, 99.6+%

GF08595119

Titanium, wire reel, 100m, diameter 1.0mm, annealed, 99.6+%

GF94150593

Titanium, wire reel, 10m, diameter 1.5mm, as drawn, 99.6+%

GF82635138

Titanium, tube, 500mm, outside diameter 6.35mm, inside diameter 5.53mm, wall thickness 0.41mm, annealed, 99.6+%

GF71020235

Titanium, wire reel, 0.5m, diameter 1.0mm, as drawn, 99.6+%

GF87716997

Titanium, rod, 100mm, diameter 16mm, as drawn, 99.99+%

GF66991179

Titanium, rod, 100mm, diameter 2mm, as drawn, 99.99+%

GF04311638

Titanium, rod, 100mm, diameter 6mm, annealed, 99.6+%

GF06082286

Titanium, microfoil, disks, 10mm, thinness 1.0μm, specific density 429μg/cm², permanent mylar 3.5μm support, 99.6+%

GF12929744

Titanium, microfoil, disks, 25mm, thinness 0.25μm, specific density 112.6μg/cm², permanent mylar 3.5μm support, 99.6+%

GF61322699

Titanium, rod, 100mm, diameter 2mm, annealed, 99.6+%

GF23916618

Titanium, microfoil, disks, 25mm, thinness 0.5μm, specific density 225.4μg/cm², permanent mylar 3.5μm support, 99.6+%

GF89377701

Titanium, tube, 100mm, outside diameter 25.4mm, inside diameter 23.62mm, wall thickness 0.89mm, annealed, 99.6+%

GF13900205

Titanium, tube, 100mm, outside diameter 0.51mm, inside diameter 0.35mm, wall thickness 0.08mm, annealed, 99.6+%

GF17705516

Titanium, rod, 200mm, diameter 3.0mm, annealed, 99.6+%

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Titanium, tube, 1000mm, outside diameter 12.7mm, inside diameter 10.9mm, wall thickness 0.9mm, annealed, 99.6+%

GF31127712

Titanium, rod, 10mm, diameter 50mm, 99.99+%

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Titanium, rod, 1000mm, diameter 3.0mm, annealed, 99.6+%

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Titanium, rod, 200mm, diameter 4mm, annealed, 99.6+%

GF34106363

Titanium, rod, 1000mm, diameter 1.5mm, annealed, 99.6+%