Abstract
Controlled release of TGF-β1 from scaffolds is an attractive mechanism to modulate the chondrogenesis of human bone marrow mesenchymal stem cells (hBMSCs) that repopulate articular cartilage defects. Here, we evaluated the ability of porous scaffolds composed of poly(ethylene oxide)-terephtalate and poly(butylene terepthalate) (PEOT/PBT) to release bioactive TGF-β1 for chondrogenesis of hBMSCs in a pellet culture model. Chondroinduction was compared with that promoted by direct addition of the recombinant factor to the culture medium. The data show a controlled release of TGF-β1 from scaffolds for at least 21 days in vitro, with ∼10% of TGF-β1 released during this period. The delivered TGF-β1 was bioactive, as confirmed by successful chondrogenic differentiation of hBMSCs monitored by morphological, histological, immunohistochemical, biochemical, and real-time reverse transcription polymerase chain reaction analyses. Third, semiquantitative histological evaluations revealed a similar pattern of chondrogenesis compared with the positive controls. Importantly, TGF-β1-loaded scaffolds allowed for a ∼700-fold upregulation of type-II collagen mRNA compared to when pellets were maintained in the presence of the soluble TGF-β1, reflected also in the highest score of immunoreactivity to type-II collagen, not significantly different from the positive controls. Likewise, aggrecan mRNA was ∼200-fold upregulated. Interestingly, most (>94%) of the glycosaminoglycan produced remaining associated with the pellets. Analysis of hypertrophic events showed no significant difference in the average total hypertrophy score compared with the positive controls. These results demonstrate the suitability of controlled TGF-β1 release from biocompatible scaffolds to promote hBMSC chondrogenesis at a physical distance and in the absence of soluble TGF-β1.
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