In vivo osteogenic evaluation of titanium implants with strontium loaded nanotubes

Abstract

Objective: To evaluate the in vivo osteogenic activity of titanium implants with strontium loaded TiO2 nanotubes (NTSr). Methods: The strontium loaded titanium nanotubes were formed on pure titanium implants through anodization and hydrothermal treatment, and the unmodified titanium (Control) and sheer TiO2 nanotubes (NT) were set to be control groups and treatment group. Inductively coupled plasma mass spectrometry (ICP-MS) was used to evaluate the Sr release at 28 days. Field emission scanning electron microscopes (FE-SEM) was used to view the micro-topography, atomic force microscope was used to exam the surface roughness, and nano-indenter was used to evaluate the hardness of three groups (n=3). Three groups of implant samples were inserted into the distal femoral metaphysis of New Zealand rabbits (n=4 at each time point). After 4 weeks and 12 weeks, samples were harvested. Micro-CT scanning, immunofluorescent and histological examinations were carried out. Results: The strontium ions could be released slowly for at least 28 days [the Sr concentration at 28 Day was (2.6±1.5) ng/ml]. NTSr coating exhibited a nanoscale tube array (the diameter was about 70 nm), and the surface roughness of implant was increased with the nanobube coating [Control (34.8±5.3) nm, NT (66.2±4.3) nm, NTSr (85.7±10.6) nm, F=37.59, P<0.001]. The surface roughness (Ra) of NT and NTSr groups was higher than the control group (P<0.05). Comparing to Control implants, NTSr implants exhibited a better osteogenic ability [the bone volume/total volume (BV/TV) value was Control (24.7±1.1)% vs. NTSr (37.7±1.9)% at 4 weeks (P<0.05), and Control (40.7±0.9)% vs. NTSr (51.9±2.1)% at 12 weeks (P<0.05)]. The fluorescent examination revealed that NTSr coating can also accelerated the generation of new bone tissue (bone tissue area% labelled by alizarin red at day 7 was Control (19.2±2.9)% vs. NT (35.4±3.7)% vs. NTSr (40.9±0.9)% (F=42.74, P<0.01). The results in the NT and NTSr group were statistically higher than that in the control group (P<0.05). Conclusions: The strontium loaded TiO2 nanotubes can enhance new bone formation around titanium implants.