To develop a porcine tooth in situ organ culture model and a novel bioreactor suitable for future studies of periodontal regeneration, including application of appropriate physiological loading.
Objective:
Method:
Twenty one porcine first molars were dissected aseptically in situ within their bony sockets. Twelve were used to optimise sterility and determine tissue viability. The remainder were used in a 4 day organ culture study in basal medium containing antibiotics (Pen/strp; 50U/ml, Amphotericin; 2.5 µg/ml, Gentamicin; 50 µg/ml). Sterility was determined for medium samples and swabs taken from sub-gingiva, gingiva, intra-radicular and surrounding bone using standard aerobic and anaerobic microbiological cultures. Tissue viability was determined at days 1 and 4 using an XTT assay. A suitable bioreactor was designed to permit longer term culture with application of mechanical loading to the tooth in situ. Diffusion characteristics were investigated using micro-CT combined with finite element modelling.
Result:
XTT assays indicated viability throughout the culture period for all tissues investigated. Clear microbiological cultures indicated maintenance of sterility within the organ culture system. The novel bioreactor showed no evidence of medium contamination after 4 days of culture. Finite element modelling indicated nutrient availability to the periodontium.
Conclusion:
A whole tooth in situ organ culture system was successfully maintained over four days in vitro. We are currently testing the organ culture model within the novel bioreactor to simulate physiological loading for future periodontal research.
Supported by WELMEC, a Centre of Excellence in Medical Engineering funded by the Wellcome Trust and EPSRC, under grant number WT 088908/Z/09/Z.