Development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells

For years, the concept of harvesting autologous Mesenchymal Stem Cells (MSCs) and applying them for regenerative medicine has been extensively studied. Numerous researchers have conducted investigations on the use of topography to stimulate in-vitro MSCs differentiation into different types of cells...

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Main Author: Liaw, Yin Siang
Other Authors: School of Chemical and Biomedical Engineering
Format: Final Year Project
Language:English
Published: 2014
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Online Access:http://hdl.handle.net/10356/60855
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-608552023-03-03T15:35:07Z Development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells Liaw, Yin Siang School of Chemical and Biomedical Engineering Kang YueJun DRNTU::Science For years, the concept of harvesting autologous Mesenchymal Stem Cells (MSCs) and applying them for regenerative medicine has been extensively studied. Numerous researchers have conducted investigations on the use of topography to stimulate in-vitro MSCs differentiation into different types of cells. However, the main limitation of previous studies on topographical factors is that the investigations has been done on typical monolayer culture system and is only limited to 1-2 layer of cells sheet formation. This poses challenges on feasibility of translation into clinical application like implantation into 3D defect site and integration to existing tissue. The objective of this project is to develop 3-Dimensional micro-patterned structures to study the effect of topographies cues to osteo- and chondrogenesis of MSCs. The scaffold was created by incorporating the topographies on the side of the wall instead of the bottom of scaffolds. Cell morphological analysis by actin cytoskeleton and nuclei counterstaining revealed that cell nuclei are flatter in scaffold without topography and more rounded in scaffold with topography. Besides that, F-actin cytoskeleton is less aligned in topographical scaffold while showing higher degree of alignment with the smoother wall. It can be deduced that MSCs exhibit higher degree of cell-cell interaction when topography is present. iii Cell proliferation test were conducted via PrestoBlue® cytotoxicity assay and results suggested that topographical environment reduces the cell viability and proliferation of MSCs while the smooth surface scaffold promotes cell viability for MSCs seeded in chondrogenic medium. qRT-PCR were conducted to obtain the gene expression level of aggrecan, alkaline phosphatase, collagen II, and collagen I, normalized to GAPDH and relative to Day 0 control. Results concluded that the topographic environment enhances chondrogenesis of MSCs under chondrogenic culture condition, but also enhances osteogenesis at the same time. The results also indicate that osteogenesis of MSCs is promoted in chondrogenic medium rather than osteogenic medium when they are under topographical influence. Bachelor of Engineering (Chemical and Biomolecular Engineering) 2014-06-02T03:41:14Z 2014-06-02T03:41:14Z 2014 2014 Final Year Project (FYP) http://hdl.handle.net/10356/60855 en Nanyang Technological University 103 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science
spellingShingle DRNTU::Science
Liaw, Yin Siang
Development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells
description For years, the concept of harvesting autologous Mesenchymal Stem Cells (MSCs) and applying them for regenerative medicine has been extensively studied. Numerous researchers have conducted investigations on the use of topography to stimulate in-vitro MSCs differentiation into different types of cells. However, the main limitation of previous studies on topographical factors is that the investigations has been done on typical monolayer culture system and is only limited to 1-2 layer of cells sheet formation. This poses challenges on feasibility of translation into clinical application like implantation into 3D defect site and integration to existing tissue. The objective of this project is to develop 3-Dimensional micro-patterned structures to study the effect of topographies cues to osteo- and chondrogenesis of MSCs. The scaffold was created by incorporating the topographies on the side of the wall instead of the bottom of scaffolds. Cell morphological analysis by actin cytoskeleton and nuclei counterstaining revealed that cell nuclei are flatter in scaffold without topography and more rounded in scaffold with topography. Besides that, F-actin cytoskeleton is less aligned in topographical scaffold while showing higher degree of alignment with the smoother wall. It can be deduced that MSCs exhibit higher degree of cell-cell interaction when topography is present. iii Cell proliferation test were conducted via PrestoBlue® cytotoxicity assay and results suggested that topographical environment reduces the cell viability and proliferation of MSCs while the smooth surface scaffold promotes cell viability for MSCs seeded in chondrogenic medium. qRT-PCR were conducted to obtain the gene expression level of aggrecan, alkaline phosphatase, collagen II, and collagen I, normalized to GAPDH and relative to Day 0 control. Results concluded that the topographic environment enhances chondrogenesis of MSCs under chondrogenic culture condition, but also enhances osteogenesis at the same time. The results also indicate that osteogenesis of MSCs is promoted in chondrogenic medium rather than osteogenic medium when they are under topographical influence.
author2 School of Chemical and Biomedical Engineering
author_facet School of Chemical and Biomedical Engineering
Liaw, Yin Siang
format Final Year Project
author Liaw, Yin Siang
author_sort Liaw, Yin Siang
title Development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells
title_short Development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells
title_full Development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells
title_fullStr Development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells
title_full_unstemmed Development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells
title_sort development of 3-dimentional micropatterned scaffolds for osteogenesis and chondrogenesis of mesenchymal stem cells
publishDate 2014
url http://hdl.handle.net/10356/60855
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