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The spreading of fibroblasts on chemically-modified collagen

Thorne, Lucy; (1995) The spreading of fibroblasts on chemically-modified collagen. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Abstract

The object of this work is to determine whether modifications in the structure of collagen, induced by its exposure in vitro to reactive aldehydes, yields a substrate that can support the growth of cultured skin fibroblasts more effectively than untreated collagen. Such studies may increase understanding of Duchenne muscular dystrophy in which the characteristic fibrosis of degenerating skeletal muscles may arise, as a secondary effect, from the peroxidation of membrane lipids. They may also contribute to the development of improved skin substitutes, based on fibroblasts embedded in a gel of chemically-modified collagen, for use in the treatment of burns. It has been reported that cell spreading and DNA synthesis are enhanced in skin fibroblasts that are cultured on collagen in which intermolecular cross-linking has been increased by incubation in vitro. I have therefore investigated both the spreading behaviour and cell division of human skin and mouse 3T3 fibroblasts on collagen that has been exposed either to malondialdehyde, which is a major product of lipid peroxidation reactions and known to cross-link collagen, or to two other reactive aldehydes (2,4-hexadienal and glutaraldehyde). Freshly isolated fibroblasts were plated on films of native and chemically-modified collagen in serum-free conditions. Both human skin and 3T3 fibroblasts exhibited a marked enhancement in cell spreading in 100 minutes at 37°C on the modified collagen preparations, by comparison with spreading on native collagen. For human skin fibroblasts, the extent of enhancement in cell spreading was proportional to the loss of collagen primary groups, with the glutaraldehyde-treated collagen showing the greatest degree of enhancement. The free tripeptide, RGD, caused an inhibition of human skin fibroblast spreading on collagen, whether in its native or modified form, indicating that this binding sequence was involved in cell spreading. My studies on thymidine incorporation into human skin fibroblasts suggest that an enhancement in the spreading of these cells on aldehyde-treated collagen did not, however, result in an increase in cell division.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: The spreading of fibroblasts on chemically-modified collagen
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest
URI: https://discovery.ucl.ac.uk/id/eprint/10098995
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