Stem cell response to a library of scaffolds with varied 3D structures was investigated. Microarray screening revealed that each type of scaffold structure induced a unique gene expression signature in primary human bone marrow stromal cells (hBMSCs). Hierarchical cluster analysis showed that treatments sorted by scaffold structure and not by polymer chemistry suggesting that scaffold structure was more influential than scaffold composition. Further, the effects of scaffold structure on hBMSC function were mediated by cell shape. Of all the scaffolds tested, only scaffolds with a nanofibrous morphology were able to drive the hBMSCs down an osteogenic lineage in the absence of osteogenic supplements. Nanofiber scaffolds forced the hBMSCs to assume an elongated, highly branched morphology. This same morphology was seen in osteogenic controls where hBMSCs were cultured on flat polymer films in the presence of osteogenic supplements (OS). In contrast, hBMSCs cultured on flat polymer films in the absence of OS assumed a more rounded and less-branched morphology. These results indicate that cells are more sensitive to scaffold structure than previously appreciated and suggest that scaffold efficacy can be optimized by tailoring the scaffold structure to force cells into morphologies that direct them to differentiate down the desired lineage.
Recognition-based assembly of micron- to nano-sized colloidal particles functionalized with DNA has generated great interest in the past decade; however, reversing the assembly process is typically achieved by thermal denaturation of the oligonucleotide duplexes. Here, we report an alternative disassembly approach at a fixed temperature using competitive hybridization events between immobilized and soluble oligonucleotide strands. Microspheres are first aggregated via primary hybridization events between immobilized DNA strands with a weak, but sufficient, affinity for partner strands to link complementary surfaces together. To reverse the aggregation process, soluble oligonucleotides are then added to competitively displace the original hybridization partners through secondary hybridization events. Using flow cytometry to quantify hybridization events and microscopy to examine DNA-mediated aggregation and redispersion, we found that the efficiency of competitive displacement is based upon (1) the difference in base pair matches between the primary and secondary target for the same probe sequence and (2) the concentration of hybridizing oligonucleotides participating in microsphere aggregation. To the best of our knowledge, this study is the first to employ DNA hybridization events to mediate reversible adhesion between colloidal particles at a fixed temperature.
This paper presents the development of a novel aluminum-filled high dielectric constant composite for embedded passive applications. Aluminum is well known as a low-cost and fast self-passivation metal. The self-passivation forms a nanoscale insulating boundary outside of the metallic spheres, which has dramatic effects on the electrical, mechanical, and chemical behaviors of the resulting composites. Influences of aluminum particle size and filler loading on the dielectric properties of composites were studied. Because of the self-passivated insulating oxide layer of fine aluminum spheres, a high loading level of aluminum can be used while the composite materials continues to be insulating. Dielectric property measurement demonstrated that, for composites containing 80 wt% 3.0 m aluminum, a dielectric constant of 109 and a low dissipation factor of about 0.02 can be achieved. The dielectric constant of epoxy-aluminum composites increased almost 30 times as compared with that of the pure epoxy matrix, which is about 3.5. Die shear tests showed that at such loading level, materials still had good processability and good adhesion toward the substrate. Bulk resistivity measurement, high-resolution transmission electron microscope (HRTEM) observation, and thermogravimetric analysis (TGA) were conducted to characterize the aluminum powders in order to understand the dielectric behavior of aluminumfilled composites. Bimodal aluminum-filled composites were also systematically studied in order to further increase the dielectric constant. Ouchiyama-Tanaka's model was used to calculate the theoretical maximum packing fraction (MPF) of bimodal systems. Based on the calculation, rheology studies were performed to find the optimum bimodal filler volume fraction ratio that led to the best packing efficiency of bimodal fillers. It was found that the viscosity of polymer composites showed a minimum at optimum bimodal filler volume fraction ratio. A high dielectric constant of 160 (@10 kHz) with a low dissipation factor of less than 0.025 was achieved with the optimized bimodal aluminum composites. The developed aluminum composite is a promising candidate material for embedded capacitor applications.
We have designed a 2-spinnerette device that can directly electrospin nanofiber scaffolds containing a gradient in composition that can be used to engineer interfacial tissues such as ligament and tendon. Two types of nanofibers are simultaneously electrospun in an overlapping pattern to create a nonwoven mat of nanofibers containing a composition gradient. The approach is an advance over previous methods due to its versatility - gradients can be formed from any materials that can be electrospun. A dye was used to characterize the 2-spinnerette approach and applicability to tissue engineering was demonstrated by fabricating nanofibers with gradients in amorphous calcium phosphate nanoparticles (nACP). Adhesion and proliferation of osteogenic cells (MC3T3-E1 murine pre-osteoblasts) on gradients was enhanced on the regions of the gradients that contained higher nACP content yielding a graded osteoblast response. Since increases in soluble calcium and phosphate ions stimulate osteoblast function, we measured their release and observed significant release from nanofibers containing nACP. The nanofiber-nACP gradients fabricated herein can be applied to generate tissues with osteoblast gradients such as ligaments or tendons. In conclusion, these results introduce a versatile approach for fabricating nanofiber gradients that can have application for engineering graded tissues.
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