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