Development of Biomaterial for 3D Bio-printed Oviduct Model

Author: Hana Ali S Alanazi

Alanazi, Hana Ali S, 2018 Development of Biomaterial for 3D Bio-printed Oviduct Model, Flinders University, College of Medicine and Public Health

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There is a continued need, particularly in developing countries, for safe, reliable and cheap contraceptives. Development of new contraceptives is very expensive and time consuming, and requires extensive animal testing. In vitro models offer a potential alternative for the early stage high-throughput screening of contraceptive candidates. The introduction of 3D bioprinting has provided a platform for the rapid manufacture of such models, however, there is still much research that needs to be conducted to identify suitable matrices that can be printed and provided an optimal environment for cells and tissues. Therefore, the aim of this study was to identify hydrogel matrices that could be used for 3D bioprinting and support the viability and function of primary-derived oviduct cells harvested from mouse oviducts. The matrices chosen for this study were agarose and carboxylated agarose (CA) hydrogels, as these gels are compatible with extrusion printing, have a high water content like the natural extracellular matrix, and are considered bioinert. Whilst this latter characteristic makes the gels cytocompatible, it also limits protein adhesion which can lead to poor cell attachment. Therefore, the gels were combined with varying amounts of the adherence proteins collagen and fibronectin to assess the effect of these biofactors on cell adhesion, viability and function when in contact with the gel surface and when encapsulated within the gel. The cytotoxicity of the candidate matrices, and their ability to support oviduct cell viability and proliferation, were initially examined using human ovarian carcinoma (OVCAR) cell lines. OVCAR cells were cultured on the surface of these gels to test their viability and ability to attach on the candidate gel surface. Whereas CA gels with or without adherence proteins supported OVCAR cell attachment and viability, no attachment was noted for any of the agarose-based gels. Subsequently, OVCAR cells were encapsulated inside the CA gels. For CA, CA+collagen and CA+fibronectin gels there was a statistically significant decrease in cell viability to ~70-80 % compared to the controls. However, CA+collagen+fibronectin gels supported OVCAR cells with > 80% viability and evidence of cell attachment. Interestingly, all CA-based gels supported the production of progesterone (P4) from the encapsulated OVCAR gels. The optimised CA+collagen+fibronectin gel was then used to successfully encapsulate primary- derived oviduct cells, which remained viable after 48 h and displayed morphological characteristics. Whilst these results suggest that CA+collagen+fibronectin gels may be promising candidates for use in a 3D bioprinted oviduct model, further work is required to identity the individual cells types present in the mixed cell population disaggregated from murine oviduct and to assess the encapsulated primary-derived oviduct cells functionality.

Keywords: Biomaterial, 3D bio-printed, Oviduct

Subject: Biotechnology thesis

Thesis type: Masters
Completed: 2018
School: College of Medicine and Public Health
Supervisor: Fiona Young