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Biomedical subjects

Aaron J Fleischman

Publications and source records attributed to Aaron J Fleischman.

10 recordsLinked to original sources
PubMed2006

Continuous flow magnetic cell fractionation based on antigen expression level.

Cell separation is important in medical and biological research and plays an increasingly important role in clinical therapy and diagnostics, such as rare cancer cell detection in blood. The immunomagnetic labeling of cells with antibodies conjugated to magnetic nanospheres gives rise to a proportional relationship between the number of magnetic nanospheres attached to the cell and the cell surface marker number. This enables the potential fractionation of cell populations by magnetophoretic mobility (MM). We exploit this feature with our apparatus, the Dipole Magnet Flow Fractionator (DMFF), which consists of an isodynamic magnetic field, an orthogonally-oriented thin ribbon of cell suspension in continuous sheath flow, and ten outlet flows. From a sample containing a 1:1 mixture of immunomagnetically labeled (label+) and unlabeled (label-) cells, we achieved an increase in enrichment of the label+ cell fraction with increasing outlet numbers in the direction of the magnetic field gradient (up to 10-fold). The total recovery of the ten outlet fractions was 90.0+/-7.7%. The mean MM of label+ cells increased with increasing outlet number by up to a factor of 2.3. The postulated proportionality between the number of attached magnetic beads and the number of cell surface markers was validated by comparison of MM measured by cell tracking velocimetry (CTV) with cell florescence intensity measured by flow cytometry.

Antigens↗
PubMed2006

Evaluation of silicon nanoporous membranes and ECM-based microenvironments on neurosecretory cells.

Understanding the interactions between microfabricated synthetic interfaces and cultured cells expressing a neuronal phenotype are critical for advancing research in the field of neural engineering such as neural recording and stimulation and neural microdevice interactions with the human brain. Here we explore the integration of these two components for therapeutic applications of neural prostheses. Microfabricated silicon nanoporous membranes were investigated for their effects on survival, proliferation, and differentiation of the well-known PC12 clonal line. Specifically, cell morphology, examined through fluorescence staining, were comparable in many respects on both silicon membrane and widely-used polystyrene culture surfaces. The attachment and differentiation of PC12 cells cultured on collagen and laminin-modified membranes and standard tissue culture surfaces were similar. Lastly, the differentiation response and tyrosine hydroxylase activity of PC12 cells embedded in a type I collagen matrix on experimental membrane substrates while exposed to NGF were significant and indistinguishable from tissue-culture polystyrene (TC-PS) surfaces. Results from this research suggest that microfabricated silicon nanoporous membranes may be useful, biocompatible permselective structures for neuroprosthetic applications and that collagen may be a useful immobilizing matrix for PC12 cells loaded in implantable macroencapsulation devices designed for the treatment of neurodegenerative disorders.

Animals↗
PubMed2006

Orthogonal-coil RF probe for implantable passive sensors.

A versatile orthogonal-coil radio frequency (RF) probe suitable for detecting the resonant frequency of miniature implantable passive sensors has been designed and tested. The probe sensitivity has been tested using printed-circuit spiral inductors of various sizes (3-15 mm) in series with discrete surface-mount capacitors designed to resonate over a range of frequencies (50-200 MHz). Close agreement between theoretical calculations and experimental results has been obtained. An equation is derived for transmit/receive (T/R) isolation that agrees with experimental measurements over the frequency range 1-500 MHz. The probe includes an additional coil to compensate for the effect of eddy currents in the human body on the probe. T/R isolation of at least 90 dB over the frequency range 1-100 MHz can be achieved when the probe is placed in close proximity to the human body.

Biosensing Techniques↗
PubMed2005

A system for micro/nano fluidic flow diagnostics.

A system for flow measurement in micro/nano fluidic components is presented. Microfabricated arrays of straight channels with noncircular cross-sections were used for flow rate measurement. The calculated flow rates in these channels were determined using a finite difference approximation method. A pneumatic pumping system was utilized to control the pressure drop across the channels and flow rates were measured by collecting the fluids on a sensitive balance. The experimental setup was validated using long narrow circular tubes that mimic the range of flow resistances characteristic of micro/nano fluidic devices. Two types of channels cross-section were investigated. The first type contained an array of channels that were approximately trapezoidal (microchannels, approximately 6.5 microm deep) in cross-section and exhibited flow rates of 27.7--119.4 micro L/min within a pressure range of 64.1--277.1 kPa (9.3--40.2 psi). The second type contained an array of channels that were approximately arc-shaped (nanochannels, approximately 600 nm deep) and generated flow rates of 0.29--0.99 micro L/min within a pressure range of 137.2--334.4 kPa (19.9--48.5 psi). The flow rates calculated by the finite difference approximation method were within 5.5% and 19.68% of the average experimental flow rates in the microchannels and nanochannels, respectively.

Computer Simulation↗
PubMed2005

Characterization of polydimethylsiloxane (PDMS) properties for biomedical micro/nanosystems.

Polydimethylsiloxane (PDMS Sylgard 184, Dow Corning Corporation) pre-polymer was combined with increasing amounts of cross-linker (5.7, 10.0, 14.3, 21.4, and 42.9 wt.%) and designated PDMS1, PDMS2, PDMS3, PDMS4, and PDMS5, respectively. These materials were processed by spin coating and subjected to common micro-fabrication, micro-machining, and biomedical processes: chemical immersion, oxygen plasma treatment, sterilization, and exposure to tissue culture media. The PDMS formulations were analyzed by gravimetry, goniometry, tensile testing, nano-indentation, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). Spin coating of PDMS was formulation dependent with film thickness ranging from 308 microm on PDMS1 to 171 microm on PDMS5 at 200 revolutions per minute (rpm). Ultimate tensile stress (UTS) increased from 3.9 MPa (PDMS1) to 10.8 MPa (PDMS3), and then decreased down to 4.0 MPa (PDMS5). Autoclave sterilization (AS) increased the storage modulus (sigma) and UTS in all formulations, with the highest increase in UTS exhibited by PDMS5 (218%). PDMS surface hydrophilicity and micro-textures were generally unaffected when exposed to the different chemicals, except for micro-texture changes after immersion in potassium hydroxide and buffered hydrofluoric, nitric, sulfuric, and hydrofluoric acids; and minimal changes in contact angle after immersion in hexane, hydrochloric acid, photoresist developer, and toluene. Oxygen plasma treatment decreased the contact angle of PDMS2 from 109 degrees to 60 degrees. Exposure to tissue culture media resulted in increased PDMS surface element concentrations of nitrogen and oxygen.

Biocompatible Materials↗
PubMed2003

Osteoblast attachment to a textured surface in the absence of exogenous adhesion proteins.

The present study investigated whether osteoblasts could attach to a culture substratum through a surface texture-dependent mechanism. Four test groups were used: (A) untextured, and three texture groups with maximum feature sizes of (B) <0.5 microm, (C) 2 microm, and (D) 4 microm, respectively. All surfaces were coated with the nonadhesive protein bovine serum albumin (BSA). Osteoblasts were allowed to adhere in serum-free medium for either 1 or 4 h, at which time nonadherent cells were removed. At 4 h, untextured surface A exhibited no cell attachment, while textured surfaces B, C, and D exhibited 9%, 32%, and 16% cell adhesion, respectively. At 16 h of incubation, adherent osteoblasts on textured surface C exhibited focal adhesion contacts and microfilament stress-fiber bundles. These results indicate that microtextured surfaces in the absence of exogenous adhesive proteins can facilitate osteoblast adhesion.

Animals↗
PubMed2002

Growth of connective tissue progenitor cells on microtextured polydimethylsiloxane surfaces.

Growth of human connective tissue progenitor cells (CTPs) was characterized on smooth and microtextured polydimethylsiloxane (PDMS) surfaces. Human bone-marrow-derived cells were cultured for 9 days under conditions promoting osteoblastic differentiation on smooth PDMS surfaces and on PDMS post microtextures that were 6 microm high and 5, 10, 20, and 40 microm in diameter, respectively. Glass tissue-culture dishes were used as controls. The number of viable cells was determined, and an alkaline phosphatase stain was used as a marker for osteoblastic phenotype. CTPs attached, proliferated, and differentiated on all surfaces. Cells on the smooth PDMS and control surfaces spread and proliferated as colonies in proximity to other cells and migrated in random directions, with cell process lengths of up to 80 microm. In contrast, cells on the PDMS post microtextures grew as sparsely distributed networks of cells, with processes, occasionally up to 300 microm, that appeared to interact with the posts. Cell counts revealed that there were fewer (50%) CTPs on the smooth PDMS surface than were on the glass control surfaces. However, there were consistently more (>144%) CTPs on the PDMS post textures than on the controls. In particular, the 10-microm-in-diameter posts (268%) exhibited a significantly (p < 0.05) greater cell number than did the smooth PDMS.

Bone Marrow Cells↗
PubMed2002

Analysis of connective tissue progenitor cell behavior on polydimethylsiloxane smooth and channel micro-textures.

Growth of human connective tissue progenitor cells (CTPs) was characterized on smooth and microtextured polydimethylsiloxane (PDMS) surfaces. Human bone marrow derived cells were cultured for nine days under conditions promoting osteoblastic differentiation on Smooth PDMS and PDMS Channel microtextures (11 microm high, 45 microm wide channels, and separated by 5 microm wide ridges). Glass tissue culture dish surfaces were used as controls. Cell numbers per colony, cell density within colonies, alignment of cells, area of colonies, and colony shapes were determined as a function of substrate surface topography. An alkaline phosphatase stain was used as a marker for osteoblastic phenotype. CTPs attached, proliferated, and differentiated on all surfaces with cell process lengths of up to 80 microm. Cells on the Smooth PDMS and control surfaces spread and proliferated as colonies in proximity to other cells and migrated in random directions creating colonies that covered significantly larger areas (0.96 and 1.05 mm(2), respectively) than colonies formed on PDMS Channel textures (0.64 mm(2)). In contrast, cells on PDMS Channel textures spread, proliferated, aligned along the channel axis, and created colonies that were more dense, and with lengths of longest colony axes that were significantly longer (3252 microm) than those on the Smooth PDMS (1265 microm) and control surfaces (1319 microm). Cells on PDMS Channel textures were aligned at an angle of 14.44 degrees relative to the channel axis, and the resulting colonies exhibited a significantly higher aspect ratio (13.72) compared to Smooth PDMS (1.57) and control surfaces (1.51).

Journal Article↗
PubMedDate not supplied

Differentiated growth of human renal tubule cells on thin-film and nanostructured materials.

Over 300,000 Americans are dependent on hemodialysis as treatment for renal failure, and kidney transplantation is limited by scarcity of donor organs. This shortage has prompted research into tissue engineering of renal replacement therapy. Existing bioartificial kidneys are large and their use labor intensive, but they have shown improved survival compared to conventional therapy in preclinical studies and an US Food and Drug Administration-approved phase 2 clinical trial. This hybrid technology will require miniaturization of hemofilters, cell culture substrates, sensors, and integration of control electronics. Using the same harvesting and isolation techniques used in preparing bioartificial kidneys for clinical use, we characterized human renal tubule cell growth on a variety of silicon and related thin-film material substrates commonly used in the construction of microelectromechanical systems (MEMS), as well as novel silicon nanopore membranes (SNMs). Human cortical tubular epithelial cells (HCTC) were seeded onto samples of single-crystal silicon, polycrystalline silicon, silicon dioxide, silicon nitride, SU-8 photoresist, SNMs, and polyester tissue culture inserts, and grown to confluence. The cells formed confluent monolayers with tight junctions and central cilia. Transepithelial resistances were similar between SNMs and polyester membranes. The differentiated growth of human tubular epithelial cells on MEMS materials strongly suggests that miniaturization of the existing bioartificial kidney will be feasible, paving the way for widespread application of this novel technology.

Biocompatible Materials↗