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Title: Optimising culture conditions for tissue engineering large articular cartilage constructs
Author: Senior, Richard
ISNI:       0000 0004 5364 7836
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2014
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Current surgical approaches to treating damage to articular cartilage, a highly specialised connective tissue, are limited in their ability to regenerate functional hyaline tissue. This has provided a driving force for the development of patient-specific, tissue engineered treatments. To date the majority of in vitro studies have focussed on engineering relatively small-dimension constructs; however justification remains for the production of large pieces of cartilage tissue. The aim of this research was therefore to investigate the potential for tissue engineering large, high quality cartilage constructs using several different culture methodologies Both small 'pin' (6 mm diameter) and large 'plate' (15 x 10 mm) constructs were successfully produced using primary bovine articular chondrocytes, a poly(glycolic acid) scaffold material and various culture conditions; static, semi-static and a rotating wall vessel (RWV) cell culture system. Small pin constructs cultured under standard static and semi-static conditions demonstrated a biochemical composition similar to that previously reported in published studies. Plate constructs cultured under static and semi-static conditions demonstrated an increased sulphated GAG and collagen type II content over their small pin counterparts, with an architecture possessing numerous lacunae and some zonal organisation. The Synthecon™ rotating wall vessel (RWV) bioreactor did not provide a suitable environment to engineer large plate constructs in standard cell culture medium. Due to their weight the constructs 'tumbled', resulting in damaged tissue with a poor quality extra cellular matrix rich in fibrous collagen type I. The design of a lightweight PTFE scaffold retention frame and the development of a dextran-modified, increased viscosity culture medium permitted the support of large constructs even at low vessel rotational RPM. The use of high viscosity culture medium in all culture environments however was found to have a detrimental impact on tissue quality, reduced mass transfer resulting in far lower matrix accumulation. It was concluded that large cartilage constructs may be produced under standard semi-static conditions that demonstrate hyaline-like features but biological quality was sacrificed. It was also concluded that an increased viscosity culture medium can demonstrate rheological properties comparable to those of synovial fluid, however in conjunction with the low-shear RWV bioreactor does not provide an ideal environment for engineering large cartilage constructs. The hydrodynamic properties of the increased viscosity culture medium could prove beneficial for the tissue engineering of articular cartilage constructs under a different bioreactor configuration.
Supervisor: Hatton, Paul ; Crawford, Aileen Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available