In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications.

HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated. In vitro differentiation capacities of HDPCs into adipogenic, osteogenic, and chondrogenic lineages were compared. Differentiation markers were analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blotting.

Our data indicated that whole HDPCs-ED, HPDCs-OG, and HDPCs-Combined could be differentiated into adipogenic, chrondrogenic, and osteogenic cell types. However, we found that the methods for isolating and culturing HDPCs influence the differentiation capacities of cells. HDPCs-OG and HDPCs-ED were preferably differentiated into adipogenic and osteogenic cells, respectively. Differentiation markers shown by RT-PCR and western blotting analysis were mostly upregulated in the treated groups compared with the control groups.

Our findings confirmed that cell populations formed by two different culture methods and the combined culture method exhibited different properties. The results of this study could provide an insight into regenerative endodontic treatment using HDPCs.

This study analyzed the difference in color caused by different thickness in enamel layer of composite resins when applied with single and layering placement technique, and evaluated if the results agreed with the shade guide from the manufacturers to verify reliability of the color matching process of the manufacturers.

For single composite resin samples, 6 mm diameter and 4 mm thickness cylindrical samples were fabricated using Ceram-X mono (DENTSPLY DeTrey) and CIE L^*a^*b^* values were measured with spectrophotometer. Same process was done for layering composite resin samples, making 3 dentinal shade samples, 4 mm thickness, for each shade using Ceram-X duo (DENTSPLY DeTrey) and enamel shade resins were layered in 2 mm thickness and CIE L^*a^*b^* values were measured. These samples were ground to 0.2 mm thickness each time, and CIE L^*a^*b^* values were measured to 1 mm thickness of enamel shade resin.

Color difference (ΔE^*) between single and layering composite resin was 1.37 minimum and 10.53 maximum when layering thicknesses were between 1 mm and 2 mm and 6 out of 10 same shade groups suggested by manufacturer showed remarkable color difference at any thickness (ΔE^* > 3.3).

When using Ceram-X mono and duo for composite resin restoration, following the manufacturer's instructions for choosing the shade is not appropriate, and more accurate information for Ceram-X duo is needed on the variation and expression of the shades depending on the thickness of the enamel.

This study examined the effect of the uncured dentin adhesives on the bond interface between the resin inlay and dentin.

Dentin surface was exposed in 24 extracted human molars and the teeth were assigned to indirect and direct resin restoration group. For indirect resin groups, exposed dentin surfaces were temporized with provisional resin. The provisional restoration was removed after 1 wk and the teeth were divided further into 4 groups which used dentin adhesives (OptiBond FL, Kerr; One-Step, Bisco) with or without light-curing, respectively (Group OB-C, OB-NC, OS-C and OS-NC). Pre-fabricated resin blocks were cemented on the entire surfaces with resin cement. For the direct resin restoration groups, the dentin surfaces were treated with dentin adhesives (Group OB-D and OS-D), followed by restoring composite resin. After 24 hr, the teeth were assigned to microtensile bond strength (µTBS) and confocal laser scanning microscopy (CLSM), respectively.

The indirect resin restoration groups showed a lower µTBS than the direct resin restoration groups. The µTBS values of the light cured dentin adhesive groups were higher than those of the uncured dentin adhesive groups (p < 0.05). CLSM analysis of the light cured dentin adhesive groups revealed definite and homogenous hybrid layers. However, the uncured dentin adhesive groups showed uncertain or even no hybrid layer.

Light-curing of the dentin adhesive prior to the application of the cementing material in luting a resin inlay to dentin resulted in definite, homogenous hybrid layer formation, which may improve the bond strength.

This in vitro study investigated whether short-term application of calcium hydroxide in the root canal system for 1 and 4 wk affects the fracture strength of human permanent teeth.

Thirty two mature human single rooted mandibular premolars in similar size and dentin thickness without decay or restorations were hand and rotary instrumented and 16 teeth vertically packed with calcium hydroxide paste and sealed coronally with caviton to imitate the endodontic procedure and the other 16 teeth was left empty as a control group. The apicies of all the samples were sealed with resin, submerged in normal saline and put in a storage box at 37℃ to mimic the oral environment. After 1 and 4 wk, 8 samples out of 16 samples from each group were removed from the storage box and fracture strength test was performed. The maximum load required to fracture the samples was recorded and data were analysed statistically by the two way ANOVA test at 5% significance level.

The mean fracture strengths of two groups after 1 wk and 4 wk were similar. The intracanal placement of calcium hydroxide weakened the fracture strength of teeth by 8.2% after 4 wk: an average of 39.23 MPa for no treatment group and 36.01 MPa for CH group. However there was no statistically significant difference between experimental groups and between time intervals.

These results suggest that short term calcium hydroxide application is available during endodontic treatment.