Chitosan/Riboflavin-modified demineralized dentin as a potential substrate for bonding.

Previous studies have suggested different approaches to modify dentin collagen for potential improvement in bonding to dentin. Here, we are proposing a new approach to reinforce dentin collagen fibrils network by chitosan as a reinforcement phase and UVA-activated riboflavin as crosslinking agent within clinically acceptable time-frame as potential substrate for bonding. The effect of modifying demineralized dentin substrates with chitosan/riboflavin, with a gradual increase in chitosan content, was investigated by SEM, nano-indentation, conventional-mechanical testing and hydroxyproline (HYP) release at collagenolytic and/or hydrolytic challenges. The resin/dentin interface morphology, immediate bond strength and short-term bond durability were also investigated using etch-and-rinse dentin adhesive. Modification with chitosan/riboflavin increased the mechanical properties, enhanced the mechanical stability of demineralized dentin substrates against hydrolytic and/or collagenolytic degradation challenges and decreased HYP release with collagenase exposure. When chitosan was added to riboflavin at 20%v/v ratio, significant improvement in bond strength at 24 h and 6 months in distilled water was found indicating the positive dual effect on bonding to dentin. With the gradual increase in chitosan content, obliteration of interfibrillar-spaces that might adversely affect bonding to dentin was found. Although it has a synergetic effect, chitosan content is crucial for any subsequent application in adhesive dentistry.

Galactosylated cellulosic sponge for multi-well drug safety testing.

Hepatocyte spheroids can maintain mature differentiated functions, but collide to form bulkier structures when in extended culture. When the spheroid diameter exceeds 200 µm, cells in the inner core experience hypoxia and limited access to nutrients and drugs. Here we report the development of a thin galactosylated cellulosic sponge to culture hepatocytes in multi-well plates as 3D spheroids, and constrain them within a macroporous scaffold network to maintain spheroid size and prevent detachment. The hydrogel-based soft sponge conjugated with galactose provided suitable mechanical and chemical cues to support rapid formation of hepatocyte spheroids with a mature hepatocyte phenotype. The spheroids tethered in the sponge showed excellent maintenance of 3D cell morphology, cell-cell interaction, polarity, metabolic and transporter function and/or expression. For example, cytochrome P450 (CYP1A2, CYP2B2 and CYP3A2) activities were significantly elevated in spheroids exposed to ß-naphthoflavone, phenobarbital, or pregnenolone-16α-carbonitrile, respectively. The sponge also exhibits minimal drug absorption compared to other commercially available scaffolds. As the cell seeding and culture protocols are similar to various high-throughput 2D cell-based assays, this platform is readily scalable and provides an alternative to current hepatocyte platforms used in drug safety testing applications.