Proteoglycan Stimulation

Project Description: This project was a mandatory requirement for the Bachelor of Engineering program at The University of Sheffield. This dissertation was supervised by Dr. Nicola Green. Additionally, the project had weekly check in’s and progressed over the entirety of my final year of study. Due to the impact of COVID-19, this project was an intensive meta-analysis instead of a wet laboratory based project.

Project Overview: This manuscript begins with outlining why the extracellular matrix is important, highlighting that collagen is predominately researched within this space, and argues that proteoglycans are severely overlooked and very important within the composition of the extracellular matrix. Although there are numerous proteoglycans this manuscript specifically focuses on biglycan, decorin, aggrecan, glypican and versican due to the representation found in literature findings.

The aims of this project:

  • To find which proteoglycans are most prevalent in 4 major tissue types: cartilage, bone, muscle and brain

  • To find where 5 identified proteoglycans: decorin, biglycan, glypican, aggrecan and versican are most prevalent

  • To determine a suitable chemical stimulant to increase proteoglycan concentration

  • To assess the efficacy of one identified chemical stimulant and its abilities to stimulate proteoglycan concentrations on different cell types

  • Further aim: Discuss the feasibility for a chemically induced proteoglycan model for the use in lab-derived ECM and TERM

Four experiments were performed, experiment 1 used mRNA expression to quantify the presence of the 5 proteoglycans within the 4 different tissue types. Experiment 2 compared 6 chemical stimulants: TGF-B, HA, TB, ANG II, BMP and FGF on their effects of stimulating proteoglycan concentration; this was quantified using [35S] Incorporation fold increase. Experimental 3 picked the chemical stimulant from experiment 2 with the highest fold increase, which was TGF-B and then compared TGF-B’s effects on different cell types. The cell types being compared were smooth muscle cells, fibroblasts and chondrocytes. This experiment was quantified using [35S] incorporation fold increase per mol of TGF-B. Experiment 4 was a hypothetical experiment that suggested from the results found in experiment 3 that smooth muscle cells are the most response per mol of TGF-B, that they would be most suitable for isolating the 5 different proteoglycans to see how each individually were stimulated from TGF-B. This experiment would be quantified using mRNA expression, and it is hypothesized that the 5 different proteoglycans being isolated would have different values after being stimulated by TGF-B.

This project required an in depth literature review, identified gaps in knowledge, extracting relevant data, processing data and determining suitability for use, methods to compare data or standardize data, statistical analysis of results, graphical representation of results, and an in depth discussion noting why this work is relevant with suggestions for future works.

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