Dose Response of MTLn3 Cells to Serial Dilutions of Arsenic Trioxide and Ionizing Radiation.

MTLn3 cells derived from mouse mammary epithelium are known to be highly malignant and are resistant to both radio- and chemo-therapy. We exposed MTLn3 cells to various doses of inorganic Arsenic trioxide (As2O3) in combination with ionizing radiation. Cells were treated with a series of As2O3 concentrations ranging from 20 µM to 1.22 nM for 8 hour, 24 hour and 48 hour periods. Post-treated cell proliferation was quantified by measuring mitochondrial activity and DNA analysis. Cells exposed to radiation and As2O3 at concentration greater than 1.25 µM showed apoptosis and radiations alone treated cells were statistically not different from the control. Hormesis was observed for As2O3 concentrations in the range of 0.078 µM to 0.625 µM while the combined chemo and radiation treatments of the cells did not affect the hormetic effect. We have demonstrated that As2O3 (in the presence and absence of ionizing radiation) in specific low concentrations induced apoptosis in the otherwise chemoresistant cancer cells. This low concentration-mediated cell death is immediately followed by a surge in cell survival. Low dosing dosimetry is highly desirable in metronomic therapy however, it has a narrow window since necrosis, hormesis, apoptosis and other dose-dependent biological processes take place in this region. Further quantifiable dosimetry is highly desired for routine clinical practice.

Neural responses to electrical stimulation on patterned silk films.

Peripheral nerve injury is a critical issue for patients with trauma. Following injury, incomplete axon regeneration or misguided axon innervation into tissue will result in loss of sensory and motor functions. The objective of this study was to examine axon outgrowth and axon alignment in response to surface patterning and electrical stimulation. To accomplish our objective, metal electrodes with dimensions of 1.5 mm × 4 cm, were sputter coated onto micropatterned silk protein films, with surface grooves 3.5 µm wide × 500 nm deep. P19 neurons were seeded on the patterned electronic silk films and stimulated at 120 mV, 1 kHz, for 45 min each day for 7 days. Responses were compared with neurons on flat electronic silk films, patterned silk films without stimulation, and flat silk films without stimulation. Significant alignment was found on the patterned film groups compared with the flat film groups. Axon outgrowth was greater (p < 0.05) on electronic films on days 5 and 7 compared with the unstimulated groups. In conclusion, electrical stimulation, at 120 mV, 1 kHz, for 45 min daily, in addition to surface patterning, of 3.5 µm wide × 500 nm deep grooves, offered control of nerve axon outgrowth and alignment.

Development path and current status of the NANIVID: a new device for cancer cell studies.

Cancer cells create a unique microenvironment in vivo that enables migration to distant organs. To better understand the tumor micro-environment, special tools and devices are required to monitor the interactions between different cell types and the effects of particular chemical gradients. Our study presents the design and optimization of a versatile chemotaxis device, the nano-intravital device (NANIVID), which consists of etched and bonded glass substrates that create a soluble factor reservoir. The device contains a customized hydrogel blend that is loaded with epidermal growth factor (EGF), which diffuses from the outlet to create a chemotactic gradient that can be sustained for many hours in order to attract specific cells to the device. A microelectrode array is under development for quantification of cell collection and will be incorporated into future device generations. Additionally, the NANIVID can be modified to generate gradients of other soluble factors in order to initiate controlled changes to the microenvironment including the induction of hypoxia, manipulation of extracellular matrix stiffness, etc. The focus of the article is to present the design and optimization of the device towards wide ranging applications of cancer cell dynamics in vitro and, ultimately, implantation for in vivo investigations.

A new chemotaxis device for cell migration studies.

This study presents the design and optimization for in vitro use of a new versatile chemotaxis device called the NANIVID (NANo IntraVital Imaging Device), developed using advanced nano/micro fabrication techniques. The device is fabricated using microphotolithographic techniques and two substrates are bonded together using a thin polymer layer creating a sealed device with one outlet. The main structure of the device consists of two Pyrex substrates: an etched chemoattractant reservoir and a top cover, with a final size of 0.2 × 2 × 3 mm. This reservoir contains a hydrogel blend with EGF which diffuses out through a small (∼9.10(3)µm(2)) outlet. This reservoir sustains a steady release of growth factor into the surrounding environment for several hours establishing a consistent concentration gradient from the device. The focus of this study was to design and optimize the new device for cell chemotaxis studies in breast cancer cells in cell culture. Our results show that we have created a flexible, cheap, miniature and autonomous chemotaxis device and demonstrate its usefulness in 2D and 3D cell culture. We also provide preliminary data for use of the device in vivo.