Abstract
Chondrocytes are frequently cultured embedded in gels that provide them with a three-dimensional environment. Nevertheless cells in these constructs cannot remodel their neighbourhood when producing their own extracellular matrix. In this work we explore 3D environments that the cells can easily remodel. For this, human mature chondrocytes were cultured in the three-dimensional environment created by chitosan microparticles whose diameter is in the order of magnitude of the cell size. Chondrocytes and microparticles suspensions were mixed and the agglomerates were cultured in static tubes in chondrogenic medium. The poor adhesion between cells and chitosan surface maintained the mobility of the ensemble. In these conditions chondrocytes are viable during the 28 days of culture. The cells produce glycosaminoglycans, S100 and collagen type II up to 14 days of culture although production of type I collagen is also noticeable.
Highlights
One of the cell sources for the regeneration of articular cartilage with tissue engineering techniques are autologous chondrocytes
Chondrocytes are frequently cultured embedded in gels that provide them with a three-dimensional environment
We explore the behaviour of human articular chondrocytes, previously expanded in monolayer, in a 3D environment formed by chitosan microparticles
Summary
One of the cell sources for the regeneration of articular cartilage with tissue engineering techniques are autologous chondrocytes. In the autologous chondrocytes implantation technique (ACI), a small number of autologous cells are obtained by enzymatic digestion of a small portion of healthy cartilage. When cells adhere to the substrate developing focal adhesion and a subsequent actin cytoskeleton they start proliferating but change their characteristic round morphology in hyaline cartilage and the expression of characteristic hyaline cartilage gens, in particular they produce type I collagen instead or in addition of type II collagen which is the predominant collagen form in the extracellular matrix of articular cartilage.[3,4] Much research effort has been dedicated to find the culture conditions that allow chondrocyte proliferation without changing its phenotype and in this sense it has been reported that the substrate material plays a role at least in retarding chondrocyte dedifferentiation.[3]
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