J Korean Acad Prosthodont.  2017 Oct;55(4):361-371. 10.4047/jkap.2017.55.4.361.

Regulation of human gingival fibroblast gene expression on microgrooves: A DNA microarray study

• Lee K
• Leesungbok R
• Ahn SJ
• Park SJ
• Lee SW

• KMID: 2393907
• DOI: http://doi.org/10.4047/jkap.2017.55.4.361

Abstract

PURPOSE
We aimed to investigate the gene expression of human gingival fibroblasts on microgroove surface using DNA microarray.
MATERIALS AND METHODS
Microgrooves were applied on grade II titanium discs to have 0/0 µm (NE0, control group), 60/10 µm (E60/10, experimental group) of respective width/depth by photolithography. The entire surface of the microgrooved Ti substrata was further acid etched and used as the two experimental groups in this study. Human gingival fibroblasts were cultured in the experimental group and the control group, and total RNA was extracted. The oligonucleotide microarray was performed to confirm the changes of various gene expression levels between experimental group and control group. Changes of gene expression level were determined at the pathway level by mapping the expression results of DNA chips, using the KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis.
RESULTS
Gene expression levels on E60/10 and NE0 were analyzed, there were 123 genes showing significant differences in expression more than 1.5 times on E60/10 microgrooved surface compared to NE0 surface, and 19 genes showing significant differences in expression more than 2 times. The KEGG pathway analysis confirmed the changes in gene expression levels under experimental conditions. Cell signaling, proliferation, and activity among the various gene expression results were identified.
CONCLUSION
Microgrooved surfaces induce gene expression changes and related cell signaling. According to the results of this study, microgrooves can be used as the surface of various biomaterials which need to improve cell activity through gene expression changes and activation of cell signaling.

Keyword

Titanium; Microgrooves; Human gingival fibroblasts; Gene expression changes

MeSH Terms

Biocompatible Materials
DNA*
Fibroblasts*
Gene Expression*
Humans*
Oligonucleotide Array Sequence Analysis*
RNA
Titanium
Biocompatible Materials
DNA
RNA
Titanium

Figure

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  Fig. 1 Fabrication of Ti substrata using photolithography.

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  Fig. 2 Field emission scanning electron microscopic images of (A) NE0 (×250), (B) E60/10 (×250) by Fe-SEM.

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  Fig. 3 A. The difference in the degree of gene expression was examined by hierarchical clustering analysis. Datas were obtained using median-center, average linkage. B is a cluster of 123 genes showing 1.5 fold changes and C is a cluster of 19 genes showing 2 fold changes.

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  Fig. 4 Changes in gene expression level in cell communication. COMP, COL5A3 were up-regulated and KRT18, COL11A1, KRTHA4, KRT7 and KRT19 were downregulated.

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  Fig. 5 Changes in gene expression level in MAPK signaling pathway. PLA2G4A, DUSP4 were up-regulated.

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  Fig. 6 Changes in gene expression level in Focal adhesion. COMP, COL5A3 were up-regulated and COL11A1, MYLK were down-regulated.

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  Fig. 7 Changes in gene expression level in Cytokine-cytokine receptor interaction. LEPR, IL7R, TNFRSF21 were down-regulated.

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  Fig. 8 Changes in gene expression level in Calcium signaling pathway. OXTR, ADRA1B, MYLK were down-regulated.

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  Fig. 9 Changes in gene expression level in JAK-STAT signaling pathway. LEPR, IL7R were down-regulated.

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  Fig. 10 Changes in gene expression level in ECM-receptor interaction. COL5A3 were up-regulated and COL11A1 were down-regulated.

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