Chih-Ang Chung

Electrical stimulation to promote osteogenesis using conductive polypyrrole films

In this study, we developed an electrical cell culture and monitoring device. Polypyrrole (PPy) films with different resistances were fabricated as conductive surfaces to investigate the effect of substrate-mediated electrical stimulation. The physical and chemical properties of the devices, as well as their biocompatibilities, were thoroughly evaluated. These PPy films had a dark but transparent appearance, on which the surface cells could be easily observed. After treating with the osteogenic medium, rat bone marrow stromal cells cultured on the PPy films differentiated into osteoblasts. The cells grown on the PPy films had up-regulated osteogenic markers, and an alkaline phosphatase activity assay showed that the PPy films accelerated cell differentiation. Alizarin red staining and calcium analysis suggested that the PPy films promoted osteogenesis. Finally, PPy films were subjected to a constant electric field to elucidate the effect of electrical stimulation on osteogenesis. Compared with the untreated group, electrical stimulation improved calcium deposition in the extracellular matrix. Furthermore, PPy films with lower resistances allowed larger currents to stimulate the surface cells, which resulted in higher levels of mineralization. Overall, these results indicated that this system exhibited superior electroactivity with controllable electrical resistance and that it can be coated directly to produce medical devices with a transparent appearance, which should be beneficial for research on electrical stimulation for tissue regeneration.

Use of biotinylated chitosan for substrate-mediated gene delivery.

To improve transfection efficiency of nonviral vectors, biotinylated chitosan was applied to complex with DNA in different N/P ratios. The morphologies and the sizes of formed nanoparticles were suitable for cell uptake. The biotinylation decreased the surface charges of nanoparticles and hence reduced the cytotoxicity. The loading capacities of chitosan were slightly decreased with the increase of biotinylation, but most of the DNA molecules were still complexed. Using different avidin-coated surfaces, the interaction between biotinylated nanoparticles to the substrate may be manipulated. The in vitro transfection results demonstrated that biotinylated nanoparticles may be bound to avidin coated surfaces, and the transfection efficiencies were thus increased. Through regulating the N/P ratio, biotinylation levels, and surface avidin, the gene delivery can be optimized. Compared to the nonmodified chitosan, biotinylated nanoparticles on biomaterial surfaces can increase their chances to contact adhered cells. This spatially controlled gene delivery improved the gene transfer efficiency of nonviral vectors and could be broadly applied to different biomaterial scaffolds for tissue engineering applications.

Hydrostatic pressure enhances mitomycin C induced apoptosis in urothelial carcinoma cells

OBJECTIVES Urothelial carcinoma (UC) of the bladder is the second most common cancer of the genitourinary system. Clinical UC treatment usually involves transurethral resection of the bladder tumor followed by adjuvant intravesical immunotherapy or chemotherapy to prevent recurrence. Intravesical chemotherapy induces fewer side effects than immunotherapy but is less effective at preventing tumor recurrence. Improvement to intravesical chemotherapy is, therefore, needed. METHODS AND MATERIALS Cellular effects of mitomycin C (MMC) and hydrostatic pressure on UC BFTC905 cells were assessed. The viability of the UC cells was determined using cellular proliferation assay. Changes in apoptotic function were evaluated by caspase 3/7 activities, expression of FasL, and loss of mitochondrial membrane potential. RESULTS Reduced cell viability was associated with increasing hydrostatic pressure. Caspase 3/7 activities were increased following treatment of the UC cells with MMC or hydrostatic pressure. In combination with 10 kPa hydrostatic pressure, MMC treatment induced increasing FasL expression. The mitochondria of UC cells displayed increasingly impaired membrane potentials following a combined treatment with 10 μg/ml MMC and 10 kPa hydrostatic pressure. CONCLUSIONS Both MMC and hydrostatic pressure can induce apoptosis in UC cells through an extrinsic pathway. Hydrostatic pressure specifically increases MMC-induced apoptosis and might minimize the side effects of the chemotherapy by reducing the concentration of the chemical agent. This study provides a new and alternative approach for treatment of patients with UC following transurethral resection of the bladder tumor.

Enhancing Immiscible Fluid Displacement in Porous Media by Capillary Pressure Discontinuities

Fluid displacement in porous media plays an important role in many industrial applications, including biological filtration, carbon capture and storage, enhanced oil recovery, and fluid transport in fuel cells. The displacement front is unstable, which evolves from smooth into ramified patterns, when the mobility (ratio of permeability to viscosity) of the displacing fluid is larger than that of the displaced one; this phenomenon is called viscous fingering. Viscous fingering increases the residual saturation of the displaced fluid, considerably impairing the efficacy of fluid displacement. It is of practical importance to develop suitable methods to improve fluid displacement. This paper presents an experimental study on applying the discontinuity of capillary pressure to improve immiscible fluid displacement in drainage for which the displacing fluid (air) wets the porous media less preferentially than does the displaced fluid (silicone oil). The concept involves using a heterogeneous packing system, where the upstream region features large pores and small capillary pressure, and the downstream region features small pores and large capillary pressure. The increase in capillary pressure prevents fingering from directly crossing the media interface, thus enhancing the displacement. The experimental apparatus was a linear cell comprising porous media between two parallel plates, and glass beads of 0.6 and 0.125 mm diameter were packed to compose the heterogeneous porous media. The time history of the finger flow was recorded using a video camera. Pressure drops over the model from the inlet to the outlet were measured to compare viscous pressure drops with capillary pressures. The results show that the fluid displacement was increased by the capillary discontinuities. The optimal displacement was determined through linear regression by adjusting the relative length of the large- and small-pore region. The results may assist in the understanding of fingering flow across the boundaries of different grain-sized bands for the gas and oil reservoir management, such as setting the relative location of the injection and production wells. The findings may also serve as a reference for industrial applications such as placing the grain bands in an adequate series to improve the displacement efficacy in biological filtration.

Osteogenic differentiation of placenta-derived multipotent cells in vitro.

OBJECTIVE Stem cells offer great potential for clinical therapeutic use because of their ability to rejuvenate and to differentiate into numerous types of cells. We isolated multipotent cells from the human term placenta that were capable of differentiation into cells of all three germ layers. MATERIALS AND METHODS We examined the ability of these placenta-derived multipotent cells (PDMCs) to differentiate into osteoblasts (OBs) or OB-like cells. The PDMCs were treated with osteogenic medium (OM) consisting of dexamethasone, β-glycerol phosphate, and ascorbic acid. At sequential time intervals (0 day, 3 days, 6 days, 9 days, and 12 days) we measured several parameters. These included alkaline phosphatase (ALP) activity, alizarin red staining (ARS) to measure calcium deposition, messenger RNA (mRNA) expressions of osteogenesis-related transcription factor (Cbfa1), and calcium coordination protein (osteocalcin). These variables were used as indicators of PDMC osteodifferentiation. RESULTS We showed that ALP activity in the early stage of differentiation and calcium deposition were both significantly increased in PDMCs after OM induction. Moreover, the Cbfa1 and osteocalcin gene expressions were upregulated. The results suggested that OM induced an osteodifferentiation potential in PDMCs. CONCLUSION PDMC-derived osteocytes provide a useful model to evaluate the mechanisms of key biomolecules and bioengineering processes.