3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning

Bibliographic Details
Main Author:	Zhao, Huizhi
Language:	English
Published:	Ohio University / OhioLINK 2018
Subjects:	Biochemistry Molecular Biology 3D cell culture Co-culture Keratinocyte Ultraviolet B irradiation DNA damage Cell transformation Nitric oxide and peroxynitrite
Online Access:	http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1531319675295094

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spelling	ndltd-OhioLink-oai-etd.ohiolink.edu-ohiou15313196752950942021-08-03T07:07:31Z 3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning Zhao, Huizhi Biochemistry Molecular Biology 3D cell culture Co-culture Keratinocyte Ultraviolet B irradiation DNA damage Cell transformation Nitric oxide and peroxynitrite Previous studies indicated that nitric oxide (NO•) plays an important role in regulation of redox balance, cell damage and apoptosis of keratinocytes upon ultraviolet B light (UVB) exposure. Since endothelial cells also release NO• for a prolonged time post-UVB, which could diffuse to adjacent cells through membrane permeation or gap junctions, we determined whether endothelial cells (HUVEC) could influence UVB-induced DNA damage and transformation of their adjacent keratinocytes (HaCaT) using a 3D polycaprolactone-based nanofiber scaffold cell co-culturing system. We chose the 3D cell culture system because recent studies had proved more similarities between 3D cell culturing models and animal models compared with 2D cell culturing. Our purpose was to build an appropriate 3D cell culturing system in which the cells could grow in multiple layers and mimic the microenvironment in tissues and solid tumors with the supply of necessary nutrients and oxygen/carbon dioxide. The polycaprolactone (biofriendly and biodegradable polymer material) scaffold was prepared using high voltage electro-spinning technique to achieve nano to micro scale polymer fibers as skeletons for cell attachment and support. In the system, HaCaT cells were co-cultured with HUVEC (co-cultured HaCaT) or with HaCaT (mono-cultured HaCaT) as the control. Our data shows that the levels of DNA damages and cyclobutane pyrimidine dimer (CPD) level along with the phosphorylation of H2AX are higher in the co-cultured than in the mono-cultured HaCaT cells post-UVB irradiation. An analysis of NO• and peroxynitrite (ONOO-) reveals that the NO• level in the co-cultured cells is increased approximately three fold than in mono-cultured HaCaT cells within one-hour post-UVB but then is reduced much quicker and remains lower in co-cultured HaCaT cells comparing to mono-cultured cells from 6-24 hours post-UVB irradiation. However, the ONOO- level is higher in the co-cultured than in the mono-cultured HaCaT cells in the early and later periods post-UVB. An analysis of nitric oxide synthases demonstrates that while expression level of iNOS is increased, the ratio of coupled/uncoupled eNOS is reduced in co-cultured HaCaT cells compared to mono-cultured HaCaT cells. Finally, our data shows that the co-cultured cells have a significantly higher transformation efficiency after repeating UVB exposure compared to mono-culture HaCaT cells. Our results suggest that endothelial cells could enhance NO•/ONOO- imbalance and promote transformation of adjacent keratinocytes. 2018-10-01 English text Ohio University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1531319675295094 http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1531319675295094 restricted--full text unavailable until 2023-09-01 This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection	NDLTD
language	English
sources	NDLTD
topic	Biochemistry Molecular Biology 3D cell culture Co-culture Keratinocyte Ultraviolet B irradiation DNA damage Cell transformation Nitric oxide and peroxynitrite
spellingShingle	Biochemistry Molecular Biology 3D cell culture Co-culture Keratinocyte Ultraviolet B irradiation DNA damage Cell transformation Nitric oxide and peroxynitrite Zhao, Huizhi 3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning
author	Zhao, Huizhi
author_facet	Zhao, Huizhi
author_sort	Zhao, Huizhi
title	3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning
title_short	3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning
title_full	3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning
title_fullStr	3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning
title_full_unstemmed	3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning
title_sort	3d cell culture model synthesized by polycaprolactone nanofiber electrospinning
publisher	Ohio University / OhioLINK
publishDate	2018
url	http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1531319675295094
work_keys_str_mv	AT zhaohuizhi 3dcellculturemodelsynthesizedbypolycaprolactonenanofiberelectrospinning
_version_	1719454212212916224

3D Cell Culture Model Synthesized By Polycaprolactone Nanofiber Electrospinning

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