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Introducing controllable 3D features into dense collagen constructs for tissue engineering applications

Alekseeva, T.; (2012) Introducing controllable 3D features into dense collagen constructs for tissue engineering applications. Doctoral thesis, UCL (University College London). Green open access

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Abstract

Plastic compression of collagen is based on unidirectional expulsion of fluid from hydrated collagen gel. The process results in dense collagen sheet, with higher density of collagen at the fluid leaving surface (FLS) than non-FLS. Compression process is completely cell-independent and at the same time cell-friendly. However, engineered tissues should replicate not only components of tissues in vivo (extracellular matrix and cells) but also their complex micro-architecture. Therefore the aim of this work was to develop collagen-based scaffolds with controllable micro-architecture for biomedical and tissue engineering applications using plastic compression (PC) of collagen. The objectives of this project were: i. to test formation of progressively opening channels in the PC collagen, ii. to investigate stable and predictable PC collagen patterning, iii. to adapt PC method in a upward-flow system as a route to process automation, iv. to investigate formation of channels using in layered PC collagen constructs. Two approaches were used in this work. Firstly, internal channels were introduced using lost fibre approach, where soluble glass fibres are incorporated in the scaffold and leave channel when dissolved. Shape and potentially progression of the channels’ opening is controlled by the shape of the template. The shape of the fibres was altered from cylindrical to conical in a controlled manner and incorporated into the PC constructs, resulting in conically-shaped channels, giving predictable internal 3D structures. The second approach relied on formation of dense collagen zone at the fluid leaving surface of the compressed collagen constructs. Formation of the densely packed collagen zone at the fluid leaving surface is essential for stable and faithful pattern formation in the process of micro-moulding. This finding has been applied in a novel upward-flow compression system to create channels using a ‘roofing’ technique. ‘Roof’ is formed by a compression of a new collagen gel on top of a patterned one; process results in open lumen channels. This appears to be due to a combination of the small dimension of the grooves in the base layer and viscosity of the collagen in the upper layer. This work demonstrates a new, previously unknown level of subtlety by which collagen fibrils can be packed and aggregated due to directional fluid flow. The outcome of this work is important for understanding pattern formation in PC collagen in vitro and potentially tissue morphogenesis in vivo. It also introduces new generation of implantable living tissue equivalents with complex micro-architecture. The multi-well compression technique has already been implemented in semi-automative working station for biomedical applications.

Type:Thesis (Doctoral)
Title:Introducing controllable 3D features into dense collagen constructs for tissue engineering applications
Open access status:An open access version is available from UCL Discovery
Language:English
UCL classification:UCL > School of Life and Medical Sciences > Faculty of Medical Sciences > Surgery and Interventional Science (Division of) > Institute of Orthopaedics and Musculoskeletal Science

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