In VitroEnhancement of SAOS-2 Cell Calcified Matrix Deposition onto Radio Frequency Magnetron Sputtered Bioglass-Coated Titanium Scaffolds

Abstract

In bone tissue engineering, bioglass coating of titanium (Ti) scaffolds has drawn attention as a method to improve osteointegration and implant fixation. In this in vitro study, bioactive glass layers with an approximate thickness of 1 microm were deposited at 200 degrees C onto a three-dimensional Ti-6Al-4V scaffold using a radio frequency (r.f.) magnetron sputtering system. After incubation with SAOS-2 human osteoblasts, in comparison with the uncoated scaffolds, the bioglass-coated scaffolds showed a twofold increase in cell proliferation (p < 0.05) up to 68.4 x 10(6), and enhanced the deposition of extracellular matrix components such as decorin, fibronectin, osteocalcin, osteonectin, osteopontin, and type-I and -III collagens (p < 0.05). Calcium deposition was twofold greater on the bioglass-coated scaffolds (p < 0.05). The immunofluorescence related to the preceding bone matrix proteins and calcium showed their colocalization to the cell-rich areas. Alkaline phosphatase activity increased twofold (p < 0.001) and its protein content was threefold higher with respect to the uncoated sample. Quantitative reverse transcriptase-polymerase chain reaction analysis revealed upregulated transcription specific for type-I collagen and osteopontin (p < 0.001). All together, these results demonstrate that the bioglass coating of the three-dimensional Ti scaffolds by the r.f. magnetron sputtering technique determines an in vitro increase of the bone matrix elaboration and may potentially have a clinical benefit.

In vitro calcified matrix deposition by human osteoblasts onto a zinc-containing bioactive glass

“…The cells were cultured in McCoy's 5A modified medium with L-glutamine and HEPES (Cambrex Bio Science, Baltimore, MD, USA), supplemented with 15% foetal bovine serum, 2% sodium pyruvate, 1% antibiotics, 10 -8 M dexamethasone, and 10 mM β-glycerophosphate (Sigma-Aldrich, St. Louis, MO, USA) (Saino et al, 2010). The osteogenic factor β-glycerophosphate was used at 10 mM concentration as previously reported in the literature (Rao et al, 2003;Gartland et al, 2005).…”

“…The samples were kept in 100% ethanol for 15 min, and then critical point-dried with CO 2 . The specimens were sputter coated with gold and observed at 250x and 1500x magnification respectively with a Leica Cambridge Stereoscan 440 microscope (Leica Microsystems, Bensheim, Germany) at 8 kV (Saino et al, 2010).…”

“…To evaluate the mitochondrial activity of the seeded cells, that is, the cell viability on the bioglass disks during the culture period, a test with 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) (Sigma-Aldrich), was performed on days 1, 8, 16, and 24 (end of the culture period) as previously reported (Saino et al, 2010). Aliquots of 200 μL were sampled, and the related absorbance values were measured at 570nm by a microplate reader (BioRad Laboratories, Hercules, CA, USA).…”

Electromagnetic Stimulation to Optimize the Bone Regeneration Capacity of Gelatin-Based Cryogels

“…The calibration curves to measure the extracted proteins were performed by an ELISA assay (24,34). The results are expressed as fg/(cell xscaffold).…”

Development and Applications of Varieties of Bioactive Glass Compositions in Dental Surgery, Third Generation Tissue Engineering, Orthopaedic Surgery and as Drug Delivery System

“…Bioactive glass and nanohydroxyapatite (BG-nHA) on titanium-alloy orthopaedic implants and surrounding bone tissue in vivo was evaluated and observed that these coatings could enhance the osteointegration of orthopaedic implant (Xie et al, 2010). Bioglass coating of the three-dimensional Ti scaffolds by the radio frequency magnetron sputtering technique determines an in vitro increase of the bone matrix elaboration and may potentially have a clinical benefit (Saino et al, 2010). Biocompatible yttrium-stabilized zirconia (YSZ) in the form of nanoparticles and bioactive Bioglass (45S5) in the form of microparticles were used to coat Ti6Al4V substrates by electrophoretic deposition with potential applications in the orthopedics (Radice et al, 2007).…”