The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications

FRICKER, CHARLOTTE

doi:https://doi.org/

E-Thesis 216 views

The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications / CHARLOTTE FRICKER

Swansea University Author: CHARLOTTE FRICKER

E-Thesis – open access under embargo until: 11th September 2029

Abstract

Collagen is the most abundant protein in the human body and as such has a variety of applications in the medical field. As society is rapidly changing with respect to ethics, there is an increasing drive to move away from the use of animal-based products. This combined with the existing issues of pa...

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Published:	Swansea, Wales, UK 2024
Institution:	Swansea University
Degree level:	Master of Research
Degree name:	MSc by Research
Supervisor:	Wright, Christopher
URI:	https://cronfa.swan.ac.uk/Record/cronfa68270

first_indexed	2025-01-30T16:02:05Z
last_indexed	2025-05-10T08:14:54Z
id	cronfa68270
recordtype	RisThesis
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spelling	2025-05-09T13:13:49.5981735 v2 68270 2024-11-14 The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications 2c6699e8c47b7cb235a32b48e03435be CHARLOTTE FRICKER CHARLOTTE FRICKER true false 2024-11-14 Collagen is the most abundant protein in the human body and as such has a variety of applications in the medical field. As society is rapidly changing with respect to ethics, there is an increasing drive to move away from the use of animal-based products. This combined with the existing issues of pathogenic transmission, immune response, and batch to batch variability of extracted collagen results in the need for a more reliable option. The process described in this research allows for the large-scale production of recombinant human collagen at a relatively low cost. Using Pichia pastoris as a host organism, plasmids containing the COL1A1 or COL1A2 genes can be integrate into the yeast cells either individually or together to produce Col(I)α1, Col(I)α2, or native triple helical collagen with a ((I)α1)2 (I)α2 composition. Combining the individual Col(I)α1 and Col(I)α2 single chains in physiological conditions has been shown to induce a similar response to extracted bovine collagen in the lactate dehydrogenase and interleukin-8 response of human umbilical vein endothelial cells indicating it may be a suitable alternative for medical devices. When both alpha chains are expressed within the same cell, the triple helical collagen produced can be seen to self-assemble into fibrils with almost native D-periodicity (84 nm) identified by atomic force microscopy. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and native gel electrophoresis were used to determine the molecular weights of the single chain collagen (100-140 kDa) and to identify the presence of triple helical molecules. The production of both human single chain and triple helical collagen with native properties at scale allows for research to be conducted which was not previously viable. This has the potential to lead to many breakthroughs in the medical field and facilitate the production of more biocompatible medical devices. E-Thesis Swansea, Wales, UK Collagen, recombinant human collagen, triple helix, recombinant protein, tissue engineering, genetic engineering 11 9 2024 2024-09-11 COLLEGE NANME COLLEGE CODE Swansea University Wright, Christopher Master of Research MSc by Research 2025-05-09T13:13:49.5981735 2024-11-14T15:54:03.9851992 Faculty of Science and Engineering School of Engineering and Applied Sciences - Biomedical Engineering CHARLOTTE FRICKER 1 Under embargo Under embargo 2025-05-09T13:13:10.3583993 Output 3361081 application/pdf E-Thesis – open access true 2029-09-11T00:00:00.0000000 Copyright: The Author, Charlotte Fricker, 2024. true eng
title	The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications
spellingShingle	The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications CHARLOTTE FRICKER
title_short	The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications
title_full	The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications
title_fullStr	The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications
title_full_unstemmed	The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications
title_sort	The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications
author_id_str_mv	2c6699e8c47b7cb235a32b48e03435be
author_id_fullname_str_mv	2c6699e8c47b7cb235a32b48e03435be_***_CHARLOTTE FRICKER
author	CHARLOTTE FRICKER
author2	CHARLOTTE FRICKER
format	E-Thesis
publishDate	2024
institution	Swansea University
college_str	Faculty of Science and Engineering
hierarchytype
hierarchy_top_id	facultyofscienceandengineering
hierarchy_top_title	Faculty of Science and Engineering
hierarchy_parent_id	facultyofscienceandengineering
hierarchy_parent_title	Faculty of Science and Engineering
department_str	School of Engineering and Applied Sciences - Biomedical Engineering{{{_:::_}}}Faculty of Science and Engineering{{{_:::_}}}School of Engineering and Applied Sciences - Biomedical Engineering
document_store_str	0
active_str	0
description	Collagen is the most abundant protein in the human body and as such has a variety of applications in the medical field. As society is rapidly changing with respect to ethics, there is an increasing drive to move away from the use of animal-based products. This combined with the existing issues of pathogenic transmission, immune response, and batch to batch variability of extracted collagen results in the need for a more reliable option. The process described in this research allows for the large-scale production of recombinant human collagen at a relatively low cost. Using Pichia pastoris as a host organism, plasmids containing the COL1A1 or COL1A2 genes can be integrate into the yeast cells either individually or together to produce Col(I)α1, Col(I)α2, or native triple helical collagen with a ((I)α1)2 (I)α2 composition. Combining the individual Col(I)α1 and Col(I)α2 single chains in physiological conditions has been shown to induce a similar response to extracted bovine collagen in the lactate dehydrogenase and interleukin-8 response of human umbilical vein endothelial cells indicating it may be a suitable alternative for medical devices. When both alpha chains are expressed within the same cell, the triple helical collagen produced can be seen to self-assemble into fibrils with almost native D-periodicity (84 nm) identified by atomic force microscopy. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and native gel electrophoresis were used to determine the molecular weights of the single chain collagen (100-140 kDa) and to identify the presence of triple helical molecules. The production of both human single chain and triple helical collagen with native properties at scale allows for research to be conducted which was not previously viable. This has the potential to lead to many breakthroughs in the medical field and facilitate the production of more biocompatible medical devices.
published_date	2024-09-11T05:25:02Z
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score	11.444473

The in vitro Assembly of Collagen Molecules and Fibrils for Advanced Tissue Engineering Applications / CHARLOTTE FRICKER

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