Developing physiological 3D in vitro tissue models for rheumatology research

Khan, Aneesah M. (2022) Developing physiological 3D in vitro tissue models for rheumatology research. PhD thesis, University of Glasgow.

Abstract

Rheumatoid arthritis is an inflammatory disease in which synovial fibroblasts maintain the persistence of inflammation within the joint, dependent on a unique pathological phenotype induced by numerous epigenetic modifications. However, knowledge of the role of these stromal cells is not completely understood. All treatments for rheumatoid arthritis are immunosuppressive drugs that target the entire immune system, resulting in many severe side effects for patients. Conventional culturing systems in 2D flat surfaces have been useful in the investigation of synovial fibroblast-dependent inflammation in rheumatoid arthritis, but inevitably lead to cellular adaptation to non-physiological plastic surfaces, that do not recapitulate the complex polarised 3D matrix where synovial fibroblasts sit in the joints. Thus, better 3D physiological models would provide a better and less artefactual understanding of the role these cells in inflammation to advance more translational research. Furthermore, understanding mechanisms through which synovial fibroblasts mediate this persistence as well as the discovery of new stromal markers in order to identify specific functional subsists of synovial fibroblasts that may be promising as disease specific therapeutic targets. This study will look at the influence of two specific 3D culture systems – polystyrene scaffold (Alvetex®) and Fibronectin pegylated hydrogels – on synovial fibroblast-dependent inflammation and their biological responses as well as the effect of the presence of matrix components on synovial fibroblast biology. The final aim of this project is to develop better physiological 3D culture systems, to obtain a better understanding of the role of fibroblasts in rheumatoid arthritis, using animal models prior to translation of these platforms to human disease.

These studies revealed that synovial fibroblasts cultured in 3D microenvironments such as Alvetex® after initially being grown in 2D, could potentially recapitulate the in vivo environment better than conventional 2D cultures, whilst collagen induced arthritis-synovial fibroblasts still present a pro-inflammatory phenotype. However interestingly, culture of the latter synovial fibroblasts in FNPEG hydrogels resulted in the cells expressing a more “remission’ like phenotype, exhibiting a less pro-inflammatory transcriptional programming. This not only provides rationale for the use of hydrogels and other 3D platforms in the study of the role of synovial fibroblasts in rheumatoid arthritis but also presents as a potential therapeutic to target synovial fibroblasts specifically.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Subjects: Q Science > QR Microbiology > QR180 Immunology
• Colleges/Schools: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Supervisor's Name: Harnett, Professor Margaret, Pineda, Dr. Miguel and Salmeron-Sanchez, Professor Manuel
• Date of Award: 2022
• Depositing User: Theses Team
• Unique ID: glathesis:2022-82838
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 29 Apr 2022 11:34
• Last Modified: 29 Apr 2022 11:41
• Thesis DOI: 10.5525/gla.thesis.82838
• URI: https://theses.gla.ac.uk/id/eprint/82838