Effect of Kelulut Honey on the Cellular Dynamics of TGFβ-Induced Epithelial to Mesenchymal Transition in Primary Human Keratinocytes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Ethical Consideration
2.2. Isolation of Primary Human Keratinocytes
2.3. Scratch Wound Healing Assay
2.4. Cellular Dynamic Parameters
2.4.1. Wound Closure Rate Analysis
2.4.2. Migration Speed Analysis
2.4.3. Migration Directionality Analysis
2.5. Cell Circularity
2.6. Immunocytochemistry
2.7. Honey Preparation
2.8. MTT Cell Viability Assay
2.9. Statistical Analysis
3. Results
3.1. TGFβ Induction Enhance In Vitro Keratinocytes Wound Closure
3.2. TGFβ Induction Increased Individual Keratinocytes Migration Velocity
3.3. TGFβ Induction Did Not Affect the Directionality of the Keratinocytes Migration
3.4. TGFβ Induction Reduced the Circularity of the Keratinocytes
3.5. TGFβ Induction Exhibit Classical EMT Markers Expression
3.6. KH Is Non-Toxic at Low Concentrations
3.7. KH Modulates TGFβ Induced Enhancement of In Vitro Keratinocytes Wound Healing
3.8. KH Modulates TGFβ Induced Enhancement of Keratinocytes Migration Velocity
3.9. KH alone Improves the Keratinocytes Migration Directionality
3.10. KH Reverses TGFβ Induced Reduction of the Keratinocytes Circularity
3.11. KH Reverses TGFβ Induced EMT Protein Expression
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ANOVA | One-way analysis of variance |
bHLH | Basic helix-loop-helix |
DMSO | Dimethyl sulfoxide |
EMT | Epithelial to mesenchymal transition |
IC50 | Half maximal inhibitory concentration |
KH | Kelulut honey |
MAPK | Mitogen-activated kinase |
MTT | 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide |
PI3K | Phosphoinositide 3-kinase |
RI | Relative intensity |
TGFβ | Tumor growth factor beta |
UKMREC | Universiti Kebangsaan Malaysia Research Ethics Committee |
ZEB | Zinc-finger E-box-binding |
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Nordin, A.; Chowdhury, S.R.; Saim, A.B.; Bt Hj Idrus, R. Effect of Kelulut Honey on the Cellular Dynamics of TGFβ-Induced Epithelial to Mesenchymal Transition in Primary Human Keratinocytes. Int. J. Environ. Res. Public Health 2020, 17, 3229. https://doi.org/10.3390/ijerph17093229
Nordin A, Chowdhury SR, Saim AB, Bt Hj Idrus R. Effect of Kelulut Honey on the Cellular Dynamics of TGFβ-Induced Epithelial to Mesenchymal Transition in Primary Human Keratinocytes. International Journal of Environmental Research and Public Health. 2020; 17(9):3229. https://doi.org/10.3390/ijerph17093229
Chicago/Turabian StyleNordin, Abid, Shiplu Roy Chowdhury, Aminuddin Bin Saim, and Ruszymah Bt Hj Idrus. 2020. "Effect of Kelulut Honey on the Cellular Dynamics of TGFβ-Induced Epithelial to Mesenchymal Transition in Primary Human Keratinocytes" International Journal of Environmental Research and Public Health 17, no. 9: 3229. https://doi.org/10.3390/ijerph17093229