Magnetic nanoparticle-loaded alginate beads for local micro-actuation of in vitro tissue constructs.

Magnetic nanoparticles (MNPs) self-align and transduce magnetic force, two properties which lead to promising applications in cell and tissue engineering. However, the toxicity of MNPs to cells which uptake them is a major impediment to applications in engineered tissue constructs. To address this problem, MNPs were embedded in millimeter-scale alginate beads, coated with glutaraldehyde cross-linked chitosan, and loaded in acellular and MDA-MB-231 cancer cell-seeded collagen hydrogels, providing local micro-actuation under an external magnetic field. Brightfield microscopy was used to assess nanoparticle diffusion from the bead. Phase contrast microscopy and digital image correlation were used to track collagen matrix displacement and estimate intratissue strain under magnetic actuation. Coating the magnetic alginate beads with glutaraldehyde-chitosan prevents bulk diffusion of nanoparticles into the surrounding microenvironment. Further, the beads exert force on the surrounding collagen gel and cells, resulting in intratissue strains of 0-10% tunable with bead dimensions, collagen density, and distance from the bead. Cells seeded adjacent to the embedded beads are subjected to strain gradients without loss of cell viability over two days culture. This study describes a simple way to fabricate crosslinked magnetic alginate beads to load in a collagen tissue construct without direct exposure of the construct to nanoparticles. The findings are significant to in vitro studies of mechanobiology in enabling precise control over dynamic mechanical loading of tissue constructs.

Development of a custom biological scaffold for investigating ultrasound-mediated intracellular delivery.

In vitro investigations of ultrasound mediated, intracellular drug and gene delivery (i.e. sonoporation) are typically carried out in cells cultured in standard plastic well plates. This creates conditions that poorly resemble in vivo conditions, as well as generating unwanted ultrasound phenomena that may confound the interpretation of results. Here, we present our results in the development of a biological scaffold for sonoporation studies. The scaffolds were comprised of cellulose fibers coated with chitosan and gelatin. Scaffold formulation was optimized for adherence and proliferation of mouse fibroblasts in terms of the ratio and relative concentration of the two constituents. The scaffolds were also shown to significantly reduce ultrasound reflections compared to the plastic well plates. A custom treatment chamber was designed and built, and the occurrence of acoustic cavitation in the chamber during the ultrasound treatments was detected; a requirement for the process of sonoporation. Finally, experiments were carried out to optimize the ultrasound exposures to minimize cellular damage. Ultrasound exposure was then shown to enable the uptake of 100nm fluorescently labeled polystyrene nanoparticles in suspension into the cells seeded on scaffolds, compared to incubation of cell-seeded scaffolds with nanoparticles alone. These preliminary results set the basis for further development of this platform. They also provide motivation for the development of similar platforms for the controlled investigation of other ultrasound mediated cell and tissue therapies.

Biomineralization of nanoscale single crystal hydroxyapatite.

The chemical and physical characteristics of nanocrystalline hydroxyapatite particles which formed during the subcutaneous implantation of crab shell in Sprague-Dawley rats were studied using selected area electron diffraction (SAED) and high resolution transmission electron microscopy (HRTEM). The initial SAED characterization evidence indicated the presence of an amorphous calcium phosphate phase. The electron dense nanophase particles which formed in the wound healing zone displayed broad diffuse rings which usually indicate a low crystalline order or amorphous phase. High resolution transmission electron microscopy (HRTEM) revealed that these mineralized regions contained discrete single crystal particles less than 5nm in size. Micrographs taken at successively higher magnifications revealed very small nanoparticles with a hexagonal arrangement of ion channels with characteristic spacing of 0.54nm and 0.23nm. This study revealed that single crystal hydroxyapatite nanoparticles consisting of only a few unit cells formed via a biomineralization directed process.

High modulus nanopowder reinforced dimethacrylate matrix composites for dental cement applications.

Results from the study of a novel, high modulus nanopowder filled resin composite are presented. This composite is developed to serve (1) as a high stiffness support to all-ceramic crowns and (2) as a means of joining independently fabricated crown core and veneer layers. Nanosized Al(2)O(3) (average particle size 47 nm) reinforcement provides stiffness across joins. Two systems are examined: Al(2)O(3) with 50:50 bis-GMA:TEGDMA monomers (ALBT) and Al(2)O(3) with pure TEGDMA (ALT). To obtain higher filler levels, surfactant is used to aid mixing and increase maximum weight percent of nanopowder filler from 72 to 80. The loading level of Al(2)O(3) has significant effects on composite properties. The elastic modulus for cured ALBT systems increases from 4.6 GPa (0 wt % filler) to 29.2 GPa (80 wt % filler). The elastic modulus for cured ALT systems increases from 3.0 GPa (0 wt % filler) to 22.9 GPa (80 wt % filler). Similarly, ALBT hardness increases from 200 MPa (0% filler) to 949 MPa (80 wt % filler), and ALT hardness increases from 93 MPa (0% filler) to 760 MPa (80 wt % filler). Our results indicate that with a generally monodispersed nanosized high modulus filler relatively high elastic modulus resin based composite cements are possible.

Scratch hardness and chipping of dental ceramics under different environments.

OBJECTIVE: The goal of this program was to identify promising environments that could efficiently minimize machining-induced damage of dental materials. METHODS: Single point abrasion (SPA) scratch testing was used on five materials to determine the scratch hardness and amount of edge chipping as functions of chemical environment, including air, water, saline and glycerol solutions. Limited testing was also done under additional environments expected to promote chemomachining effects via crack growth promotion or debris removal. A conical diamond indenter and a conventional tungsten carbide machining tool were used in the scratch tests. One-way ANOVA analysis was used to determine statistical differences among the variables. RESULTS: There was a consistent trend across materials that the water and saline yielded the lowest values of scratch hardness, air the next lowest, and the tests performed in glycerol yielded the highest hardness values. The measured hardness values using the conical diamond tool in the glycerol environments were about twice the hardness values measured under water and saline solutions. Environmental effects on chipping were minimal, but a linear relationship between load and per cent chipping was determined for the WC tool within the 10-50 N test range. The choice of scratch tool strongly affected scratch hardness and chipping tendency. SIGNIFICANCE: The chemical environment had an effect on machining characteristics, but the effects were more dependent on tool interactions rather than material specific properties. As a result, it may not be possible to utilize a particular single environment to substantially improve the damage response of dental materials to machining operations. Improvements in damage resistance can be environmentally obtained, but only for shallow cuts (finishing operations).