Development and characterisation of ECM-rich tissue surrogates using synthetic crowders


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Project Components

In this section, each student (e.g. A, B and C) should provide a background, objectives and identify specific tasks and proposed methodology for their individual project component.
Each student should use a separate section, e.g. Student A should complete Section 2.1, Student B should complete Section 2.2 etc…
2.1 Subtopic A: Insert Title Here
2.1.1 Preliminary Literature Review (A)
The main body text should be Size 12pt, Times New Roman, single line spacing and justified throughout.
2.1.2 Key Individual Objectives (A)
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2.1.3 Identification of Tasks and Methodology (A)

The main body text should be Size 12pt, Times New Roman, single line spacing and justified throughout.

2.1 Subtopic A: Hyaluronic acid as a macromolecular crowding agent in the development and characterisation of ECM-rich tissue surrogates
2.1.1 Hyaluronic acid plays a role regulating cell differentiation, migration, angiogenesis and inflammation responses [4]. Hyaluronic acid can be described as a non-sulfated, naturally occurring non-protein glycosaminoglycan, with distinct physico-chemical properties, produced by synoviocytes, fibroblasts, and chondrocytes [5]. Based on the use of Hyaluronic acid as a macromolecular crowding agent for production of cell-derived matrices in a literature from 2019, Hyaluronic acid was used as a crowding agent with Human dermal fibroblasts. Human dermal fibroblasts were cultures with 0%, 0.05% and 0.5% high molecular weight Hyaluronic Acid for 3, 7 or 14 days. The control used for this experiment was Ficoll 70/400 [2].
On day 14 results showed that SDS-Page, Sircol and hydroxyproline assays determined that 0.05% Hyaluronic acid treated cultures presented a greater higher mean collagen deposition than the control. Fluorescent immunostaining of ECM proteins did not show much difference in ECM production in both Hyaluronic acid and the ficoll 70/400 control. The use of Raman imaging showed that Hyaluronic acid increased ECM deposition in Human Dermal Fibroblasts [2].
2.1.2 Key Individual Objectives (A)
Understanding how crowding affects tissue surrogates. Developing a deeper understanding of how Natural non-sulphated polysaccharides ( Ficoll, hyaluronic acid) and Natural sulphated polysaccharides ( carrageenan) macromolecular crowding agents and how it impacts the ECM.
2.1.3 Identification of Tasks and Methodology (A)
I will gather and analyse data determining if hyaluronic acid is a suitable macromolecular crowder for ECM-rich tissue surrogates.
Steps involved in the process of analysis include the Live/ Death assay for 4, 7 or 10 days using AlamarBlue Viability assay. Metabolic activity for 4, 7 or 10 days. Proliferation of cultures, Immunocytochemistry and SDS-PAGE which is used to separate proteins based on their molecular weight.
This data will be compared with a control and the mechanical properties of the crowder will be analysed.


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