Expression of dentine sialophosphoprotein in mouse nasal cartilage.

Objective: Dentine sialophosphoprotein (DSPP) was initially thought to be unique for dentine formation during tooth development, whilst recent reports have shown a much broader expression pattern such as in bone, periodontium and inner ear. Our goal was to explore its expression and potential impact during early nasal cartilage formation in comparison with tooth development.

Study Design: We investigated DSPP expression in the nasal cartilage by immunohistochemistry and in situ hybridisation.

[Study of cbfalpha1 on the expression of extracellular matrix proteins in dental papilla cells induced by BMP-2 in vitro].

Purpose: To evaluate the function of cbfalpha1 on BMP-2 signaling to extracellular matrix proteins in dental papilla cells in vitro.

Methods: RT-PCR and Western blot were performed to detect the expression of ALP, OC, ON, OPN, BSP, DMP-1 and DSPP in cultured dental papilla cells induced by 200ng/mL BMP-2 and/or down-regulated by cbfalpha1 antisense technology, the results were analysed with SPSS 11.0 software package.

[The effect of core binding factor alpha1 on transcriptional regulation of mouse dentin sialophosphoprotein].

Purpose: To investigate the effect of core binding factor alpha1 (cbfalpha1) on transcriptional regulation of mouse dentin sialophosphoprotein (DSPP) gene.

Methods: The MDPC-23 cells and the segment of nt -2475bp to +53bp were chosen. After co-transfected, the MDPC-23 cells were measured for luciferase activity using the dual luciferase reporter assay system.

[Cloning and characterization of genes differentially expressed in human dental pulp cells and gingival fibroblasts].

Objective: To study the biological properties of human dental pulp cells (HDPC) by cloning and analysis of genes differentially expressed in HDPC in comparison with human gingival fibroblasts (HGF).

Methods: HDPC and HGF were cultured and identified by immunocytochemistry. HPDC and HGF subtractive cDNA library was established by PCR-based modified subtractive hybridization, genes differentially expressed by HPDC were cloned, sequenced and compared to find homogeneous sequence in GenBank by BLAST.

[Generation of transgenic mouse of dentin sialoprotein and transgene expression analysis].

Purpose: To generate the transgenic mouse model of DSP and perform transgene expression analysis by RT-PCR.

Methods: Plasmid pcDNA3.1-CX was constructed by substituting promoter cbeta-actin for CMV promoter of pcDNA3.

[Immortalized human odontoblast-like cell line expressed dentin extracellular matrix in vitro].

Objective: To determine whether dentin matrix proteins were expressed by the human odontoblast-like cell line hTERT-hOd-1 in vitro.

Methods: Collagen type I, bone sialoprotein (BSP), dentin matrix protein 1 (DMP1) and the marker for odontoblast, dentin sialophosphoprotein (DSPP) and dentin sialoprotein (DSP) were detected in these cells by immunohistochemistry, RT-PCR and in situ hybridization. During being cultured in mineralizing medium for 5 weeks, the secretion of OC and activity of ALP were measured once a week.

[IRF6 gene mutation analysis in a van Der Woude syndrome family in Henan province].

Purpose: To investigate IRF6 gene mutation in a van Der Woude syndrome (VWS) family in Henan province.

Methods: PCR and DNA sequencing was employed to detect the mutation of IRF6.Secondary construction transformation analysis was performed using PIX-Protein Identification software.

[Generation of dspp-LacZ transgenic mouse founders].

Purpose: To establish a transgenic mouse founders in which the expression of LacZ was directed by a dentin sialophosphoprotein-specific promoter.

Methods: The DSPP-specific promoter was obtained by PCR and confirmed by sequencing, and the transgenic plasmid, pTN-DPM-LacZ, was constructed by subcloning the DSPP-specific promoter and LacZ-encoding sequence into one vector. The linearized transgenic plasmid was microinjected into the male pronucleus of the zygotes, and the microinjected zygotes were implanted to recipient pseudopregnant mice.

[Cloning and sequencing of the upstream of mouse dentin sialophosphoprotein promoter].

Objective: To clone and sequence the upstream of mouse dentin sialophosphoprotein promoter.

Methods: Genomic DNA was obtained from Balb/c mouse blood. The upstream of mouse dentin sialophosphoprotein promoter segments was obtained by PCR.

[Cbfa1 induces the expression of the mineral-related proteins in human dental papilla cells].

Objective: To explicit whether the expression of the mineral-related proteins is regulated by cbfa1 in human dental papilla cells.

Methods: Human dental papilla cells were cultured in vitro and transfected with pcDNA3-cbfa1 recombinant plasmids. After selected with G418 sulfate, a cell clone named PC-3, which could stably express the cbfa1 mRNA and protein, was proved by PCR and western blot.