Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release
This thesis presents the development and evaluation of two applications for scaffolds in the field of liver tissue engineering. In the first study a poly (D,L lactic acid) (PDLLA) scaffold is used as a three-dimensional template for hepatocyte–hepatic stellate cell (HSC) co-culture. To enhance PDLLA...
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| Format: | Thesis (University of Nottingham only) |
| Language: | English |
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2009
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| Online Access: | https://eprints.nottingham.ac.uk/12556/ |
| _version_ | 1848791525335498752 |
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| author | Hammond, John Stotesbury |
| author_facet | Hammond, John Stotesbury |
| author_sort | Hammond, John Stotesbury |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | This thesis presents the development and evaluation of two applications for scaffolds in the field of liver tissue engineering. In the first study a poly (D,L lactic acid) (PDLLA) scaffold is used as a three-dimensional template for hepatocyte–hepatic stellate cell (HSC) co-culture. To enhance PDLLA ligand binding capacity scaffolds are surface modified using allylamine plasma deposition and treatment with NaOH. Primary adult rat hepatocytes and HSC are then seeded onto these scaffolds and cultured in static conditions. Scanning electron microscopy (SEM) is used to assess mono-culture and co-culture morphology whilst synthetic and cytochrome P450 function are measured using albumin and testosterone assays.
The second study explores the potential for intrahepatic growth factor and extracellular matrix (ECM) delivery from a biodegradable polymer scaffold to promote liver growth and to enhance regeneration. The study is undertaken in rats. The scaffold design and implantation technique are first piloted in a short survival study. Hepatocyte growth factor (HGF), epidermal growth factor (EGF), fibroblast growth factor (FGF)1, FGF2 and liver derived ECM (L-ECM) are then loaded into poly(lactic-co-glycolic acid) (PLGA) + 5% poly(ethylene glycol) (PEG) scaffolds and implanted into normal and partially hepatectomised liver. Implant morphology is assessed by micro-CT reconstruction. Growth factor bioactivity and release are confirmed by in vitro profiling. Liver growth and volume redistribution are assessed by liver weight analysis. Parenchymal injury and function are quantified by measuring serum aspartate aminotransferase (AST) & bilirubin. 5-bromodeoxyuridine (BrdU) inclusion & MIB-5 immunohistochemistry (IHC) are used to identify hepatocyte and non-parenchymal cell proliferation. Liver-scaffold interaction is characterised by H&E and Masson’s trichrome staining. Non-parenchymal cell migration is assessed by ED-1 and desmin IHC. All histology is then subjected to image analysis. |
| first_indexed | 2025-11-14T18:29:54Z |
| format | Thesis (University of Nottingham only) |
| id | nottingham-12556 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T18:29:54Z |
| publishDate | 2009 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-125562025-02-28T11:19:57Z https://eprints.nottingham.ac.uk/12556/ Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release Hammond, John Stotesbury This thesis presents the development and evaluation of two applications for scaffolds in the field of liver tissue engineering. In the first study a poly (D,L lactic acid) (PDLLA) scaffold is used as a three-dimensional template for hepatocyte–hepatic stellate cell (HSC) co-culture. To enhance PDLLA ligand binding capacity scaffolds are surface modified using allylamine plasma deposition and treatment with NaOH. Primary adult rat hepatocytes and HSC are then seeded onto these scaffolds and cultured in static conditions. Scanning electron microscopy (SEM) is used to assess mono-culture and co-culture morphology whilst synthetic and cytochrome P450 function are measured using albumin and testosterone assays. The second study explores the potential for intrahepatic growth factor and extracellular matrix (ECM) delivery from a biodegradable polymer scaffold to promote liver growth and to enhance regeneration. The study is undertaken in rats. The scaffold design and implantation technique are first piloted in a short survival study. Hepatocyte growth factor (HGF), epidermal growth factor (EGF), fibroblast growth factor (FGF)1, FGF2 and liver derived ECM (L-ECM) are then loaded into poly(lactic-co-glycolic acid) (PLGA) + 5% poly(ethylene glycol) (PEG) scaffolds and implanted into normal and partially hepatectomised liver. Implant morphology is assessed by micro-CT reconstruction. Growth factor bioactivity and release are confirmed by in vitro profiling. Liver growth and volume redistribution are assessed by liver weight analysis. Parenchymal injury and function are quantified by measuring serum aspartate aminotransferase (AST) & bilirubin. 5-bromodeoxyuridine (BrdU) inclusion & MIB-5 immunohistochemistry (IHC) are used to identify hepatocyte and non-parenchymal cell proliferation. Liver-scaffold interaction is characterised by H&E and Masson’s trichrome staining. Non-parenchymal cell migration is assessed by ED-1 and desmin IHC. All histology is then subjected to image analysis. 2009-07-22 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/12556/1/J_Hammond_e-thesis.pdf_%28cut_version%29.pdf Hammond, John Stotesbury (2009) Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release. PhD thesis, University of Nottingham. |
| spellingShingle | Hammond, John Stotesbury Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release |
| title | Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release |
| title_full | Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release |
| title_fullStr | Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release |
| title_full_unstemmed | Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release |
| title_short | Scaffolds for liver tissue engineering: in vitro co-culture & in vivo release |
| title_sort | scaffolds for liver tissue engineering: in vitro co-culture & in vivo release |
| url | https://eprints.nottingham.ac.uk/12556/ |