Mechanical loading of chondrocytes embedded in 3D constructs: in vitro methods for assessment of morphological and metabolic response to compressive strain.
Mechanical loading of chondrocytes in 3D constructs has been used to investigate mechanotransduction and its potential for stimulating tissue-engineered cartilage repair. This chapter describes the preparation of 3D agarose or alginate constructs seeded with isolated chondrocytes and specific test rigs for applying gross compressive strain to individual constructs on a confocal microscope or for longer term compression of constructs cultured within an incubator. Experimental methods are described to quantify the level of cell deformation and the elaboration of extracellular matrix. The chapter thus provides an introduction to the experimental techniques used to examine chondrocyte mechanotransduction and downstream cell function.
IRC in Biomedical Materials and Medical Engineering Division, Department of Engineering, Queen Mary College, University of London, London, UK.
David A
DA
Lee
Martin M
MM
Knight
D000464:Alginates / Q000737:chemistry; D000818:Animals; D019149:Bioreactors; D002417:Cattle; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D019245:Compressive Strength; D005452:Fluoresceins / Q000032:analysis; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D040542:Mechanotransduction, Cellular; D011509:Proteoglycans / Q000032:analysis / Q000096:biosynthesis; D012685:Sepharose / Q000737:chemistry; D023822:Tissue Engineering
D000464:Alginates; D005452:Fluoresceins; D006603:Hexuronic Acids; D011509:Proteoglycans; C085925:calcein AM; D020723:Glucuronic Acid; D012685:Sepharose