Novel nanocomposite scaffold based on gelatin/PLGA-PEG-PLGA hydrogels embedded with TGF-β1 for chondrogenic differentiation of human dental pulp stem cells in vitro.

• Nanocomposite hydrogels consisted of gelatin and PLGA-PEG-PLGA copolymer were prepared and characterized. • For the whole time of the experiment, the gelatin/PLGA-PEG-PLGA composite hydrogel displayed biocompatibility with human dental pulp stem cells. • As regards cell viability, ECM production as well as differentiation of hDPSCs into chondrocyte-like cells, gelatin/PLGA-PEG-PLGA hydrogel demonstrated to be the promising carrier for hDPSCs. • The gelatin/PLGA-PEG-PLGA composite hydrogels exhibited the modulus comparative to articular cartilage and desirable biocompatibility to articular chondrocytes. In the current study, a novel nanocomposite hydrogel scaffold comprising of natural-based gelatin and synthetic-based (poly D, L (lactide-co-glycolide) -b- poly (ethylene glycol)-b- poly D, L (lactide-co-glycolide) (PLGA-PEG-PLGA) triblock copolymer was developed and loaded with transforming growth factor- β1 (TGF-β1). Synthesized scaffolds' chemical structure was examined by 1H NMR and ATR-FTIR. Scanning electron microscopy (SEM) confirmed particle size and morphology of the prepared nanoparticles as well as the scaffolds. The morphology analysis revealed a porous interconnected structure throughout the scaffold with a pore size dimension of about 202.05 µm. The swelling behavior, in vitro degradation, mechanical properties, density, and porosity were also evaluated. Phalloidin/DAPI staining was utilized for confirming the extended cytoskeleton of the chondrocytes. Alcian blue staining was conducted to determine cartilaginous matrix sulfated glycosaminoglycan (sGAG) synthesis. Eventually, over a period of 21 days, a real-time RT-PCR analysis was applied to measure the mRNA expression of chondrogenic marker genes, type-II collagen, SOX 9, and aggrecan, in hDPSCs cultured for up to 21 days to study the influence of gelatin/PLGA-PEG-PLGA-TGF-β1 hydrogels on hDPSCs. The findings of the cell-encapsulating hydrogels analysis suggested that the adhesion, viability, and chondrogenic differentiation of hDPSCs improved by gelatin/PLGA-PEG-PLGA-TGF-β1 nanocomposite hydrogels. These data supported the conclusion that gelatin/PLGA-PEG-PLGA-TGF-β1 nanocomposite hydrogels render the features that allow the in vitro functionality of encapsulated hDPSCs and hence can contribute the basis for new effective strategies for the treatment of cartilage injuries. [ABSTRACT FROM AUTHOR]