J Dent Rehabil Appl Sci.  2020 Sep;36(3):145-157. 10.14368/jdras.2020.36.3.145.

Analysis of cell survival genes in human gingival fibroblasts after sequential release of trichloroacetic acid and epidermal growth factor using the nano-controlled release system

• Cho JY
• Leesungbok R
• Lee SW

• KMID: 2512109
• DOI: http://doi.org/10.14368/jdras.2020.36.3.145

Abstract

Purpose
This study was to determine the possible effects of trichloroacetic acid (TCA) and epidermal growth factor (EGF) through cell survival genes of the PI3K-AKT signaling pathway when applying an hydrophobically modified glycol chitosan (HGC)-based nanocontrolled release system to human gingival fibroblasts in oral soft tissue regeneration.
Materials and Methods
An HGC-based nano-controlled release system was produced, followed by the loading of TCA and EGF. The group was divided into control (CON), TCA-loaded nano-controlled release system (EXP1), and the TCA- and EGF- individually loaded nano-controlled release system (EXP2).A total for 29 genes related to the PI3K-AKT signaling pathway were analyzed after 48h of culture in human gingival fibroblasts. Real-time PCR, 1- way ANOVA and multiple regression analysis were performed.
Results
Cell survival genes were significantly upregulated in EXP1 and EXP2. From multiple regression analysis, ITGB1 was determined to be the most influential factor for AKT1 expression.
Conclusion
The application of TCA and EGF through the HGC-based nano-controlled release system can up-regulate the cell survival pathway.

Keyword

human gingival fibroblast; trichloroacetic acid; epidermal growth factor; PI3K-AKT pathway; cell survival

Figure

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  Fig. 1 Principle of HGC-based TCA- and EGF- loaded nano-controlled release system. The hydrophilic cationic amine group (-NH2) of chitosan was combined with hydrophobic cholanic acid. Hydrophobic cholanic acid formed nuclei in water, while hydrophilic cationic chitosan formed the outer shell. The anionic TCA and EGF were loaded on the chitosan shell through ion bonding.

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  Fig. 2 Real-time PCR analysis results of Growth factor (IGF1R, EFGR, IRS1), Toll-like receptor signaling pathway (TLR2, TLR4, Rac1), B cell receptor (BCR) signaling pathway (BCR, SYK, CD19, PHF11), JAK/STAT signaling pathway (IL2RG, JAK1, JAK2, JAK3), Focal adhesion associated genes (ITGA5, ITGB1, PTK2). One-way ANOVA (***: P < 0.001, **: P < 0.01, *: P < 0.05).

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  Fig. 3 Real-time PCR analysis results of Chemokine signaling pathway associated genes (GNB1, GNG2) and other genes directly involved in AKT production (PDPK1, HSP90AA1, Cdc37, mTOR, TCL1A). One-way ANOVA (***: P < 0.001).

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  Fig. 4 Real-time PCR analysis results of PI3K genes (PI3KCA, PI3KCB, PI3KCD, PI3KCG). One-way ANOVA (***: P < 0.001).

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  Fig. 5 Real-time PCR analysis results of AKT1. One-way ANOVA (***: P < 0.001).

Reference

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