Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells

Numerous types of single cells in in-vitro cultures have been studied in tissue engineering, but the study on direct paracrine interactions between heterotypic cells population is lacking. Co-culture approach establishes an excellent atmosphere to study these interactions. The objective of this i...

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Main Author:	Allah, Nasar Um Min
Format:	Thesis
Language:	English
Published:	2018
Subjects:	RC Internal medicine
Online Access:	http://eprints.usm.my/46589/1/Dr.%20Nasar%20Um%20Min%20Allah-24%20pages.pdf
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spelling	my-usm-ep.465892020-06-21T08:29:04Z Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells 2018-11 Allah, Nasar Um Min RC Internal medicine Numerous types of single cells in in-vitro cultures have been studied in tissue engineering, but the study on direct paracrine interactions between heterotypic cells population is lacking. Co-culture approach establishes an excellent atmosphere to study these interactions. The objective of this in-vitro experimental study was to determine the effects of fibroblast and platelet-derived growth factor ((FGF-2 and PDGF-BB) in a co-culture of human gingival fibroblasts (HGFs) and human umbilical vein endothelial cells (HUVECs). To this end, the medium for the establishment of monolayer and co-culture of these cells were first optimised. Thereafter, the optimal concentrations of these growth factors were determined in a monolayer and then in a co-culture medium by assessing the cell viability using MTT assay. Next, gene expression analysis for fibroblast and angiogenic biomarkers was assessed using realtime RT-PCR to study the stimulatory effect of these growth factors by using 6 wellplate with transwell inserts. Afterwards, statistical analysis of the results was performed using one-way ANOVA and Kruskal-Wallis test with p < 0.05 considered statistically significant. Results of cell viability assay showed that the effect of FGF-2 on HGF was dose-dependent and was optimum at a concentration of 5 ng/ml (p =0.001), while that of PDGF-BB on HUVEC was optimum at a concentration of 20 ng/ml (p =0.004). The stimulatory effect of FGF-2 and PDGF-BB on HGF and HUVEC was supported by the real-time RT-PCR results which showed that there is a significant upregulation (p < 0.05) of gene biomarkers in the treatment group of bothcells after three days of co-culture experiment, compared to control group. Therefore, it is concluded that there is the possibility of a synergistic effect of these two growth factors on a co-culture of HGF and HUVEC which were suggestive of a proangiogenic activity. 2018-11 Thesis http://eprints.usm.my/46589/ http://eprints.usm.my/46589/1/Dr.%20Nasar%20Um%20Min%20Allah-24%20pages.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Perubatan
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	RC Internal medicine
spellingShingle	RC Internal medicine Allah, Nasar Um Min Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells
description	Numerous types of single cells in in-vitro cultures have been studied in tissue engineering, but the study on direct paracrine interactions between heterotypic cells population is lacking. Co-culture approach establishes an excellent atmosphere to study these interactions. The objective of this in-vitro experimental study was to determine the effects of fibroblast and platelet-derived growth factor ((FGF-2 and PDGF-BB) in a co-culture of human gingival fibroblasts (HGFs) and human umbilical vein endothelial cells (HUVECs). To this end, the medium for the establishment of monolayer and co-culture of these cells were first optimised. Thereafter, the optimal concentrations of these growth factors were determined in a monolayer and then in a co-culture medium by assessing the cell viability using MTT assay. Next, gene expression analysis for fibroblast and angiogenic biomarkers was assessed using realtime RT-PCR to study the stimulatory effect of these growth factors by using 6 wellplate with transwell inserts. Afterwards, statistical analysis of the results was performed using one-way ANOVA and Kruskal-Wallis test with p < 0.05 considered statistically significant. Results of cell viability assay showed that the effect of FGF-2 on HGF was dose-dependent and was optimum at a concentration of 5 ng/ml (p =0.001), while that of PDGF-BB on HUVEC was optimum at a concentration of 20 ng/ml (p =0.004). The stimulatory effect of FGF-2 and PDGF-BB on HGF and HUVEC was supported by the real-time RT-PCR results which showed that there is a significant upregulation (p < 0.05) of gene biomarkers in the treatment group of bothcells after three days of co-culture experiment, compared to control group. Therefore, it is concluded that there is the possibility of a synergistic effect of these two growth factors on a co-culture of HGF and HUVEC which were suggestive of a proangiogenic activity.
format	Thesis
qualification_level	Master's degree
author	Allah, Nasar Um Min
author_facet	Allah, Nasar Um Min
author_sort	Allah, Nasar Um Min
title	Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells
title_short	Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells
title_full	Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells
title_fullStr	Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells
title_full_unstemmed	Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells
title_sort	effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Perubatan
publishDate	2018
url	http://eprints.usm.my/46589/1/Dr.%20Nasar%20Um%20Min%20Allah-24%20pages.pdf
_version_	1747821691848884224

Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells

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