Analysis of gene and protein expression in healthy and carious tooth pulp with cDNA microarray and two-dimensional gel electrophoresis
ABSTRACT Complementary DNA (cDNA) microarray and two-dimensional (2-D) gel electrophoresis, combined with mass spectrometry, enable simultaneous analysis of expression patterns of thousands of genes, but their use in pulp biology has been limited. Here we compared gene and protein expression of pulp tissues from sound and carious human teeth using cDNA microarray and 2-D gel electrophoresis to evaluate their usefulness in pulp biology research and to identify the genes with changes in carious teeth. The cDNA microarray revealed several differentially expressed genes and genes with a high expression in both tissues. These genes have various functions, e.g. effects on vascular and nerve structures, inflammation, and cell differentiation. Variability between cDNA hybridizations indicates that the overall gene expression pattern may vary significantly between individual teeth. The 2-D gel electrophoresis revealed no change between healthy and diseased tissue. The identification of 96 proteins in the pulp tissue revealed none of the gene products with corresponding high/different mRNA expression in cDNA microarray. Interestingly, we detected also a hypothetical protein (putative nucleoside diphosphate kinase), and present therefore the first evidence for the existence of this protein. Even though the methods reveal potentially important gene expression, they may currently have only limited value in in vivo pulp biology research.
- SourceAvailable from: Bernardo A. Petriz
[Show abstract] [Hide abstract]
- "Endodontic infections have a complex polymicrobial etiology characterized by complex interrelationships between endodontic microorganisms and host immune-inflammatory response (Nair, 1997). However, Pääkkö nen et al. (2005) did not find differences in protein expression between healthy and mildly carious tooth pulp, despite the detection of 96 proteins in both groups, including one novel protein (putative nucleoside diphosphate kinase) by 2-DE and MALDI-TOF-MS and some differences in gene expression detected by cDNA microarray (Pääkkö nen et al., 2005). "
ABSTRACT: Despite all the dental information acquired over centuries and the importance of proteome research, the cross-link between these two areas only emerged around mid-nineties. Proteomic tools can help dentistry in the identification of risk factors, early diagnosis, prevention, and systematic control that will promote the evolution of treatment in all dentistry specialties. This review mainly focuses on the evolution of dentistry in different specialties based on proteomic research and how these tools can improve knowledge in dentistry. The subjects covered are an overview of proteomics in dentistry, specific information on different fields in dentistry (dental structure, restorative dentistry, endodontics, periodontics, oral pathology, oral surgery, and orthodontics) and future directions. There are many new proteomic technologies that have never been used in dentistry studies and some dentistry areas that have never been explored by proteomic tools. It is expected that a greater integration of these areas will help to understand what is still unknown in oral health and disease. J. Cell. Physiol. 228: 2271-2284, 2013. © 2013 Wiley Periodicals, Inc.Journal of Cellular Physiology 12/2013; 228(12):2271-84. DOI:10.1002/jcp.24410 · 3.84 Impact Factor
[Show abstract] [Hide abstract]
- "Many studies using cDNA microarray assay have been done: comparative studies between healthy pulp and caries infected pulp, comparative studies of tooth and bone marrow cells, comparative studies of fibroblast from healthy gingiva and inflammatory gingiva and gene expression comparison between osteoblast and fibroblast.9-12 The development of microarray assay for large-scale analysis of mRNA gene expression makes it possible to search key molecules systemincally.13,14 "
ABSTRACT: We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization. Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data. Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated. Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.08/2012; 37(3):142-8. DOI:10.5395/rde.2012.37.3.142
[Show abstract] [Hide abstract]
- "It also reveals potential biomarkers for diagnosis and estimation of pulp tissue defense potential, and enables identification of potential target proteins for treatment . Previously large-scale screenings of expression differences between healthy and carious pulp tissue and between native pulp tissue and odontoblasts using microarray have been performed   . "
ABSTRACT: Transforming growth factor-beta 1 (TGF-beta1) is the most extensively studied growth factor in dentin-pulp complex, with pleiotropic effects on pulp response and healing. Our main objective was to analyze the expression profile of pulp tissue and odontoblasts, and the effects of TGF-beta1 on these profiles in cultured human pulp and odontoblasts with a specific interest in the anti- and pro-inflammatory cytokines. For that purpose, pulps and odontoblasts were cultured for different time periods, and microarray was performed to both cultured and native samples. Of cytokines, various interleukins (IL) were confirmed by RT-PCR, and in +/- TGF-beta1 treated pulps also by antibody array. Pro-inflammatory IL-7, -12alpha and -16 mRNAs were detected in native pulp. The expression levels of pro-inflammatory IL-1alpha, -1beta, -6 and -8 were clearly induced after TGF-beta1 treatment, while no anti-inflammatory cytokines were induced. Of all pulpal interleukins analyzed IL-6 and -8 were present at the highest levels in conditioned pulp tissue media. In native odontoblasts pro-inflammatory IL-6 and -7 mRNAs were detected, and in cultured odontoblasts pro-inflammatory IL-8 mRNA showed over 20-fold transient induction after TGF-beta1 treatment. Our results demonstrate that TGF-beta1 is a potent regulator of pro-inflammatory responses and defensive reactions in dentin-pulp complex.Cytokine 11/2007; 40(1):44-51. DOI:10.1016/j.cyto.2007.08.003 · 2.66 Impact Factor