Gelatin microparticles aggregates as three-dimensional scaffolding system in cartilage engineering

Abstract

A three-dimensional (3D) scaffolding system for chondrocytes culture has been produced by agglomeration of cells and gelatin microparticles with a mild centrifuging process. The diameter of the microparticles, around 10 μ, was selected to be in the order of magnitude of the chondrocytes. No gel was used to stabilize the construct that maintained consistency just because of cell and extracellular matrix (ECM) adhesion to the substrate. In one series of samples the microparticles were charged with transforming growth factor, TGF-β1. The kinetics of growth factor delivery was assessed. The initial delivery was approximately 48 % of the total amount delivered up to day 14. Chondrocytes that had been previously expanded in monolayer culture, and thus dedifferentiated, adopted in this 3D environment a round morphology, both with presence or absence of growth factor delivery, with production of ECM that intermingles with gelatin particles. The pellet was stable from the first day of culture. Cell viability was assessed by MTS assay, showing higher absorption values in the cell/unloaded gelatin microparticle pellets than in cell pellets up to day 7. Nevertheless the absorption drops in the following culture times. On the contrary the cell viability of cell/TGF-β1 loaded gelatin microparticle pellets was constant during the 21 days of culture. The formation of actin stress fibres in the cytoskeleton and type I collagen expression was significantly reduced in both cell/gelatin microparticle pellets (with and without TGF-β1) with respect to cell pellet controls. Total type II collagen and sulphated glycosaminoglycans quantification show an enhancement of the production of ECM when TGF-β1 is delivered, as expected because this growth factor stimulate the chondrocyte proliferation and improve the functionality of the tissue.

Evaluation of the chondrogenic differentiation of mesenchymal stem cells on hybrid biomimetic scaffolds

“…These results indicated that GEL has accelerated the proliferation and differentiation of MSCs cells. This positive effect of GEL on cell proliferation has been reported in the literature . Another cause might be an RGD sequence of GEL, as a denatured collagen, that improved cell adhesion.…”

Targeted Delivery of Bioactive Molecules for Vascular Intervention and Tissue Engineering

“…Microsphere (MS)-mediated growth factor delivery has been used for years to mature and differentiate many engineered tissues, including cartilage ( Solorio et al, 2010 ), bone ( Wang et al, 2009 ), and stem cell aggregates ( Carpenedo et al, 2009 ; Solorio et al, 2010 ; Bratt-Leal et al, 2013 ). MS incorporation alone can increase tissue strength ( Solorio et al, 2012b ; Dikina et al, 2015 ), oxygen diffusion ( Hayashi and Tabata, 2011 ; Tajima and Tabata, 2013 ), cell viability ( Garcia Cruz et al, 2013 ; Tajima and Tabata, 2013 ), and uniformity of matrix deposition ( Solorio et al, 2012a ). However, their application in vascular tissue engineering has been limited.…”

Gelatin Microsphere for Cartilage Tissue Engineering: Current and Future Strategies

“…At this phase, the preparation of any drugs or biological agents is done and later mixed with the gelatin aqueous solution. For protein encapsulation by gelatin, the preheated gelatin solution is cooled down to body temperature to avoid protein denaturation, or by applying the protein solution onto the GM [ 42 , 43 ]. The gelatin solution is mixed with preheated oil and stirred to obtain the water in oil emulsion.…”