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
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...
| 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 |
| 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 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. |
|---|---|
| Keywords: |
Collagen, recombinant human collagen, triple helix, recombinant protein, tissue engineering, genetic engineering |
| College: |
Faculty of Science and Engineering |