Effect of microgrooves and fibronectin conjugation on the osteoblast marker gene expression and differentiation

PURPOSE: To determine the effect of fibronectin (FN)-conjugated, microgrooved titanium (Ti) on osteoblast differentiation and gene expression in human bone marrow-derived mesenchymal stem cells (MSCs). MATERIALS AND METHODS: Photolithography was used to fabricate the microgrooved Ti, and amine functionalization (silanization) was used to immobilize fibronectin on the titanium surfaces. Osteoblast differentiation and osteoblast marker gene expression were analyzed by means of alkaline phosphatase activity assay, extracellular calcium deposition assay, and quantitative real-time PCR. RESULTS: The conjugation of fibronectin on Ti significantly increased osteoblast differentiation in MSCs compared with non-conjugated Ti substrates. On the extracellular calcium deposition assays of MSCs at 21 days, an approximately two-fold increase in calcium concentration was observed on the etched 60-microm-wide/10-microm-deep microgrooved surface with fibronectin (E60/10FN) compared with the same surface without fibronectin (E60/10), and a more than four-fold increase in calcium concentration was observed on E60/10FN compared with the non-etched control (NE0) and etched control (E0) surfaces. Through a series of analyses to determine the expression of osteoblast marker genes, a significant increase in all the marker genes except type I collagen alpha1 mRNA was seen with E60/10FN more than with any of the other groups, as compared with NE0. CONCLUSION: The FN-conjugated, microgrooved Ti substrate can provide an effective surface to promote osteoblast differentiation and osteoblast marker gene expression in MSCs.

The effect of IDS (immediate dentin sealing) on dentin bond strength under various thermocycling periods

PURPOSE: The purpose of this study was to find out the effect of immediate dentin sealing (IDS) on bond strength of ceramic restoration under various thermocycling periods with DBA (dentin bonding agent system). MATERIALS AND METHODS: Fifty freshly extracted human mandibular third molars were divided into 5 groups (1 control and 4 experimental groups) of 10 teeth. We removed enamel layer of sound teeth and embedded them which will proceed to be IDS, using All Bond II. A thermocycling was applied to experimental groups for 1, 2, 7, 14 days respectively and was not applied to control group. IPS Empress II for ceramic was acid-etched with ceramic etchant (9.5% HF) and silane was applied. Each ceramic disc was bonded to specimens with Duo-link, dual curable resin cement by means of light curing for 100 seconds. After the cementation procedures, shear bond strength measurement and SEM analysis of the fractured surface were done. The data were analyzed with a one-way ANOVA and Tukey multiple comparison test (alpha=.05). RESULTS: There were no statistically significant differences between 4 experimental groups and control group, however the mean value started to decrease in group 7d, and group 14d showed the lowest mean bond strength in all groups. Also, group 7d and 14d showed distinct exposed dentin and collapsed hybrid layer was observed in SEM analysis. CONCLUSION: In the present study, it can be concluded that ceramic restorations like a laminate veneer restoration should be bonded using resin cement within one week after IDS procedure.