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Postgraduate Thesis: Two-photon photochemical crosslinking-based fabrication of protein microstructures
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TitleTwo-photon photochemical crosslinking-based fabrication of protein microstructures
 
AuthorsXu, Jinye
徐金叶
 
Issue Date2011
 
PublisherThe University of Hong Kong (Pokfulam, Hong Kong)
 
AbstractOne of the challenges in tissue engineering is to fabricate scaffolds which can mimic the natural microenvironments of cells. In a cell niche, biophysical and mechanical cues are crucial factors influencing cell functions. Given the complexity of natural extracellular matrix (ECM) engineered ECMs providing controllable biophysical and mechanical cues are appealing both in enhancing the understanding of cell-matrix interaction and in controlling cell fates in vitro. The ultimate goal of our study is to establish a platform as an engineered ECM by fabricating customized solid protein microstructures from solution using two-photon photochemical crosslinking, a novel laser-based freeform fabrication technique. In this study, protein structures varying from submicron lines, 2D micropatterns and microporous matrices, to 3D micropillars were successfully fabricated, demonstrating freeform fabrication capability with two-photon photochemical crosslinking. Two-photon fluorescent imaging and scanning electron microscope (SEM)-based microstructural characterization revealed that power, scan speed, total exposure time and concentrations of protein (bovine serum albumin) and photosensitizer (rose Bengal) in the solution were crucial processing parameters in this fabrication technique. Quantitative imaging analysis showed that porosity of protein matrices was highly dependent on processing parameters including power, scan speed, number of cycles in time series scan and protein concentrations in the solution. An atomic force microscopy (AFM)-based step change nano-compression test was used to measure the reduced elastic modulus of 3D viscoelastic protein micro-pillars fabricated, as a pilot study. Microporous protein matrices and 3D micropillar arrays fabricated with two-photon photochemical crosslinking can be used as engineered ECM for future study in cell-ECM interactions.
 
AdvisorsChan, BP
Cheung, KMC
 
DegreeMaster of Philosophy
 
SubjectExtracellular matrix proteins.
Solid freeform fabrication.
 
Dept/ProgramMechanical Engineering
 
DC FieldValue
dc.contributor.advisorChan, BP
 
dc.contributor.advisorCheung, KMC
 
dc.contributor.authorXu, Jinye
 
dc.contributor.author徐金叶
 
dc.date.hkucongregation2012
 
dc.date.issued2011
 
dc.description.abstractOne of the challenges in tissue engineering is to fabricate scaffolds which can mimic the natural microenvironments of cells. In a cell niche, biophysical and mechanical cues are crucial factors influencing cell functions. Given the complexity of natural extracellular matrix (ECM) engineered ECMs providing controllable biophysical and mechanical cues are appealing both in enhancing the understanding of cell-matrix interaction and in controlling cell fates in vitro. The ultimate goal of our study is to establish a platform as an engineered ECM by fabricating customized solid protein microstructures from solution using two-photon photochemical crosslinking, a novel laser-based freeform fabrication technique. In this study, protein structures varying from submicron lines, 2D micropatterns and microporous matrices, to 3D micropillars were successfully fabricated, demonstrating freeform fabrication capability with two-photon photochemical crosslinking. Two-photon fluorescent imaging and scanning electron microscope (SEM)-based microstructural characterization revealed that power, scan speed, total exposure time and concentrations of protein (bovine serum albumin) and photosensitizer (rose Bengal) in the solution were crucial processing parameters in this fabrication technique. Quantitative imaging analysis showed that porosity of protein matrices was highly dependent on processing parameters including power, scan speed, number of cycles in time series scan and protein concentrations in the solution. An atomic force microscopy (AFM)-based step change nano-compression test was used to measure the reduced elastic modulus of 3D viscoelastic protein micro-pillars fabricated, as a pilot study. Microporous protein matrices and 3D micropillar arrays fabricated with two-photon photochemical crosslinking can be used as engineered ECM for future study in cell-ECM interactions.
 
dc.description.naturepublished_or_final_version
 
dc.description.thesisdisciplineMechanical Engineering
 
dc.description.thesislevelmaster's
 
dc.description.thesisnameMaster of Philosophy
 
dc.identifier.hkulb4717922
 
dc.languageeng
 
dc.publisherThe University of Hong Kong (Pokfulam, Hong Kong)
 
dc.relation.ispartofHKU Theses Online (HKUTO)
 
dc.rightsThe author retains all proprietary rights, (such as patent rights) and the right to use in future works.
 
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License
 
dc.source.urihttp://hub.hku.hk/bib/B47179223
 
dc.subject.lcshExtracellular matrix proteins.
 
dc.subject.lcshSolid freeform fabrication.
 
dc.titleTwo-photon photochemical crosslinking-based fabrication of protein microstructures
 
dc.typePG_Thesis
 
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<item><contributor.advisor>Chan, BP</contributor.advisor>
<contributor.advisor>Cheung, KMC</contributor.advisor>
<contributor.author>Xu, Jinye</contributor.author>
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<date.issued>2011</date.issued>
<description.abstract>&#65279;One of the challenges in tissue engineering is to fabricate scaffolds which can mimic the natural microenvironments of cells. In a cell niche, biophysical and mechanical cues are crucial factors influencing cell functions. Given the complexity of natural extracellular matrix (ECM) engineered ECMs providing controllable biophysical and mechanical cues are appealing both in enhancing the understanding of cell-matrix interaction and in controlling cell fates in vitro. 



The ultimate goal of our study is to establish a platform as an engineered ECM by fabricating customized solid protein microstructures from solution using two-photon photochemical crosslinking, a novel laser-based freeform fabrication technique. 



In this study, protein structures varying from submicron lines, 2D micropatterns and microporous matrices, to 3D micropillars were successfully fabricated, demonstrating freeform fabrication capability with two-photon photochemical crosslinking.



Two-photon fluorescent imaging and scanning electron microscope (SEM)-based microstructural characterization revealed that power, scan speed, total exposure time and concentrations of protein (bovine serum albumin) and photosensitizer (rose Bengal) in the solution were crucial processing parameters in this fabrication technique. Quantitative imaging analysis showed that porosity of protein matrices was highly dependent on processing parameters including power, scan speed, number of cycles in time series scan and protein concentrations in the solution. 



An atomic force microscopy (AFM)-based step change nano-compression test was used to measure the reduced elastic modulus of 3D viscoelastic protein micro-pillars fabricated, as a pilot study. 



Microporous protein matrices and 3D micropillar arrays fabricated with two-photon photochemical crosslinking can be used as engineered ECM for future study in cell-ECM interactions.</description.abstract>
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