title: Monitoring of metabolite gradients in tissue-engineered constructs.
abstract: At present, the assessment of developing tissue-engineered constructs is almost always carried out destructively using biochemical or histological methods to determine cell number, viability and tissue growth throughout the construct. Since many of these experiments are long, taking weeks or even months to complete, simple and readily applicable non-destructive methods of monitoring changes in cell metabolism, viability and tissue deposition within the construct would be invaluable; such methods could point out adverse responses during the early stages of culture. Here, we describe the use of microdialysis for detecting local changes in cellular metabolism within a tissue-engineered construct. Three-dimensional constructs consisting of bovine articular chondrocytes entrapped in an alginate gel were cultured in a bioreactor for two weeks. Glucose and lactate were monitored by microdialysis, as the major nutrient and metabolite, respectively. Concentration gradients within the construct were evident, with the highest lactate concentrations in the construct centre. The local lactate concentration was a measure of cellular metabolic activity, decreasing as cellular activity fell and increasing as cellular activity was stimulated. Nutrient starvation and cell death in the construct centre could be readily detected in constructs deliberately cultured under adverse conditions. The results show that probe measurements can give an early warning of inappropriate local metabolic changes. Such information during the growth of tissue-engineered constructs would allow either corrective action or else an early end to an unsuccessful test.
authors.affiliation: Department of Engineering Science, University of Oxford, Parks Road, Oxford OX1 3PJ, UK. olga.boubriak@physiol.ox.ac.uk
authors.forename: Olga A
authors.initials: OA
authors.lastname: Boubriak
authors.forename: Jill P G
authors.initials: JP
authors.lastname: Urban
authors.forename: Zhanfeng
authors.initials: Z
authors.lastname: Cui
mesh_terms: D000464:Alginates / Q000737:chemistry*; D000818:Animals; D019149:Bioreactors; D002417:Cattle; D019902:Chondrocytes / Q000502:physiology*; D005947:Glucose / Q000737:chemistry / Q000378:metabolism*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D020100:Hydrogels / Q000737:chemistry; D019344:Lactic Acid / Q000737:chemistry / Q000378:metabolism*; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020100:Hydrogels; D019344:Lactic Acid; D020723:Glucuronic Acid; D005947:Glucose