BBSRC EASTBIO DTP: Making Connections: Building in vitro models of tissue interfaces using 3D bioprinting technologies

This project in a collaborative and interdisciplinary venture between Dr Paxton, an expert in tissue engineering, and Dr Murray, an expert in molecular analysis of the neuromuscular system.

There are many regions in the human body where two different tissues join together. For example, in the musculoskeletal system, the soft tissues of tendon and ligament join to the hard tissue of bone though a unique, carefully constructued connecting region, which has an important job in protecting both tissues from damage1,2. As a functional connection, the microanatomy of the bone-tendon/ligament interface (or enthesis) is important to understand since tendon/ligament injury is common in sports. When a tendon/ligament is ruptured, and surgery required to re-attach the soft tissue to bone, the normal tissue connection structure is lost, and unorganised scar tissue is produced, leaving the surgical interface weak and prone to further injury.

This project aims to apply novel techniques in tissue engineering to develop a reproducible co-culture method whereby the cells from two different musculoskeletal tissues (e.g. bone and tendon) can be grown in close apposition. Then, we aim to apply cutting-edge molecular techniques to investigate what happens when cells from ajoining tissues grow together during development and after injury. By doing so, we aim to develop a tool whereby future investigations into treatment options can be explored with the ultimate goal of improving patient outcomes following musculoskeletal injury. To accopmlish this, we will employ the novel technology of extrusion-based 3D bioprinting3, where cells are encaspuslated within a polymer ‘bioink’ and ‘printed’ with high precision in culture plates. The Paxton Lab owns a 3D bioprinter with a duel printhead functionality, which will enable the printing of duel cell types (e.g. bone and tendon cells) in polymers in a variety of different orientations, modelling a 3D interfacial region. Once 3D co-culture constructs have been bioprinted, we will profile the molecular changes occuring when cells interact. RNA will be extracted from co-cultures at regular time points following co-culture creation, and RNAseq analysis will be used to profile the evolving transcriptome as cells interact. Bioinformatic analysis will then be used to identify key cellular pathways and signalling cascades which mediate the interactions between bone and tendon cells as they form an rudimentary enthesis. This will provide an insight into the developmental pathways to investigate further in the co-culture systems and help to formulate a novel model system to study interfacial development and repair.

The key experimental techniques involved in this project are;
• 2D/3D cell culture,
• 3D bioprinting,
• histology
• immunohistochemistry,
• RNAseq
• Bioinformatic analysis of transcriptional data

Training will also be provided in experimental design, data collection, data analysis and presentation skills.

Applications:
Completed application form along with your curriculum vitae should be sent to our PGR student team at [Email Address Removed]

References:
Please send the reference request form to two referees. Completed forms for University of Edinburgh College of Medicine and Veterinary Medicine project should be returned to [Email Address Removed] by the closing date: 5th December 2018.

It is your responsibility to ensure that references are provided by the specified deadline.

Download application and reference forms via:
https://www.ed.ac.uk/roslin/postgraduate/bbsrc-eastbio-dtp