Biomedical subjects

Ketul C Popat

Publications and source records attributed to Ketul C Popat.

9 recordsLinked to original sources

PubMed2007

Osteogenic differentiation of marrow stromal cells cultured on nanoporous alumina surfaces.

A major goal in orthopedic biomaterials research is to design implant surfaces, which will enhance osseointegration in vivo. Several microscale as well as nanoscale architectures have been shown to significantly affect the functionality of bone cells i.e., osteoblasts. In this work, nanoporous alumina surfaces fabricated by a two-step anodization process were used. The nanostructure of these surfaces can be controlled by varying the voltage used for anodization process. Marrow stromal cells were isolated from mice and seeded on nanoporous and amorphous (control) alumina surfaces. Cell adhesion, proliferation, and viability were investigated for up to 7 days of culture. Furthermore, the cell functionality was investigated by calcein staining. The cells were provided with differentiation media after 7 days of culture. The alkaline phosphatase (ALP) activity and matrix production were quantified using a colorimetric assay and X-ray photoelectron spectroscopy (XPS) for up to 3 weeks of culture (2 weeks after providing differentiation media). Further, scanning electron microscopy (SEM) was used to investigate osteoblast morphology on these nanoporous surfaces. Over the 3-week study, the nanoporous alumina surfaces demonstrated approximately 45% increase in cell adhesion, proliferation, and viability, 35% increase in ALP activity, and 50% increase in matrix production when compared with the control surfaces.

Alkaline Phosphatase↗

PubMed2006

Nanostructured surfaces for bone biotemplating applications.

A major goal of orthopedic biomaterials research is to design better surface chemistries and configurations to control behavior of bone cells such as osteoblasts. Nanostructured architecture significantly affects the response of several cell lines. In this work, nanostructured surfaces were prepared by vapor liquid solid growth of silicon nanowires from size-controlled gold colloid catalysts deposited on fused silica substrates. The lengths and surface densities of the nanowires were varied to assess the effect of these parameters on bone cell response. Osteoblasts were seeded on nanowire surfaces to investigate both short-term adhesion and proliferation and long-term functionality and matrix production. Cell adhesion and proliferation were characterized using a standard 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay and cell counting for up to 4 days of culture. The total protein content, alkaline phosphatase activity, and matrix production were quantified using standard colorimetric assays for up to 4 weeks of culture. Matrix production was also characterized by measuring surface concentrations of calcium and phosphorus using X-ray photoelectron spectroscopy. Further, scanning electron microscopy was used to investigate osteoblast morphology on nanostructured surfaces. Over the 4-week study, the nanostructured surfaces demonstrated improved osteoblast adhesion and proliferation and increased alkaline phosphatase activity and matrix production compared to non-nanostructured control surfaces.

Alkaline Phosphatase↗

PubMed2005

Modeling of RGDC film parameters using X-ray photoelectron spectroscopy.

Immobilization of peptides on surfaces is a common method to investigate biological response to biomaterials for the development of improved tissue engineering constructs. Peptide immobilization can be achieved by either physical adsorption or covalent attachment on the surface. In this work, the RGDC peptide was covalently immobilized to alumina substrate for investigation of bone cell response. An understanding of RGDC film formation is necessary for further biological characterization. Thus, X-ray photoelectron spectroscopy was used to chemically characterize these films. These data were further utilized to calculate RGDC film thickness, density, and coverage, which are essential for understanding bone cell interactions with the underlying films and substrate.

Adsorption↗

PubMed2005

Fabrication and evaluation of nanoporous alumina membranes for osteoblast culture.

An understanding of osteoblast response to surface topography is essential for successful bone tissue engineering applications. Alumina has been extensively used as a substrate for bone tissue constructs. However, current techniques do not allow precise surface topography and orientation of the material. In this research, a two-step anodization process was optimized for the fabrication of nanoporous alumina membranes with uniform pore dimension and distribution. The anodization voltage can be varied to create nanoporous alumina membranes with pore sizes ranging from 30 to 80 nm in diameter. The impact of the nanoscale pores on osteoblast response was studied by evaluating cell adhesion, morphology, and matrix production. Scanning electron microscopy and atomic force microscopy were used to characterize the nanoporous alumina membranes. Osteoblast adhesion and morphology were investigated using scanning electron microscopy images and matrix production was characterized using energy dispersive spectroscopy. This research combined the advantages of using alumina, a material with proven biocompatibility and current orthopedic clinical applications, and incorporated porous features on the nanoscale which have been reported to improve osteoblast response.

Aluminum Oxide↗

PubMed2005

Peptide-immobilized nanoporous alumina membranes for enhanced osteoblast adhesion.

Bone tissue engineering requires the ability to regulate cell behavior through precise control over substrate topography and surface chemistry. Understanding of the cellular response to micro-environment is essential for biomaterials and tissue engineering research. This research employed alumina with porous features on the nanoscale. These nanoporous alumina surfaces were modified by physically adsorbing vitronectin and covalently immobilizing RGDC peptide to enhance adhesion of osteoblasts, bone-forming cells. X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) were used to characterize the modified nanoporous alumina surface. Survey and high-resolution C1s scans suggested the presence of RGDC and vitronectin on the surface and SEM images confirmed the pores were not clogged after modification. Cell adhesion on both unmodified and modified nanoporous alumina was compared using fluorescence microscopy and SEM. RGDC was found to enhance osteoblast adhesion after 1 day of culture and matrix production was visible after 2 days. Cell secreted matrix was absent on unmodified membranes for the same duration. Vitronectin-adsorbed surfaces did not show significant improvement in adhesion over unmodified membranes.

Adsorption↗

PubMed2004

Influence of nanoporous alumina membranes on long-term osteoblast response.

A major goal of bone tissue engineering is to design better scaffold configuration and materials to better control osteoblast behavior. Nanoporous architecture has been shown to significantly affect cellular response. In this work, nanoporous alumina membranes were fabricated by a two-step anodization method to investigate bone cell response. Osteoblasts were seeded on nanoporous alumina membranes to investigate both short-term adhesion and proliferation and long-term functionality and matrix production. Cell adhesion and proliferation were characterized using a standard MTT assay and cell counting. The total protein content was measured after cell lysis using the BCA assay. Matrix production was characterized in terms of surface concentrations of calcium and phosphorous, components of bone matrix, using X-ray photoelectron spectroscopy (XPS). The results from nanoporous alumina membranes were compared with those of amorphous alumina, aluminum, commercially available ANOPORE and glass. Results indicate improved osteoblast adhesion and proliferation and increased matrix production after 4 weeks of study.

Aluminum↗

PubMed2004

Surface modification of nanoporous alumina surfaces with poly(ethylene glycol).

Nanoporous alumina surfaces have a variety of applications in biosensors, biofiltration, and targeted drug delivery. However, the fabrication route to create these nanopores in alumina results in surface defects in the crystal lattice. This results in inherent charge on the porous surface causing biofouling, that is, nonspecific adsorption of biomolecules. Poly(ethylene glycol) (PEG) is known to form biocompatible nonfouling films on silicon surfaces. However, its application to alumina surfaces is very limited and has not been well investigated. In this study, we have covalently attached PEG to nanoporous alumina surfaces to improve their nonfouling properties. A PEG-silane coupling technique was used to modify the surface. Different concentrations of PEG for different immobilization times were used to form PEG films of various grafting densities. X-ray photoelectron spectroscopy (XPS) was used to verify the presence of PEG moieties on the alumina surface. High-resolution C1s spectra show that with an increase in concentration and immobilization time, the grafting density of PEG also increases. Further, a standard overlayer model was used to calculate the thickness of PEG films formed using the XPS intensities of the Al2p peaks. The films formed by this technique are less than 2.5 nm thick, suggesting that such films will not clog the pores which are in the range of 70-80 nm.

Adsorption↗

PubMed2004

Neurotransmitter analog tethered to a silicon platform for neuro-BioMEMS applications.

The design of chemically well-defined, machinable surfaces containing neuroactive molecules offers potential for fundamental neuroscience and clinical neural engineering applications. Here we report the assembly and characterization of silicon platforms containing a tethered form of muscimol. Muscimol, an analog of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), is a potent agonist at postsynaptic GABA(A) and GABA(C) receptors. Surfaces were assembled using covalent avidin conjugation to silanized silicon followed by high-affinity avidin-biotin binding of a biotinylated derivative of muscimol (muscimol-biotin). Contact angle measurements, ellipsometry, and X-ray photoelectron spectroscopy (XPS) were conducted to characterize the wettability, thickness, and chemical composition of progressively deposited surface layers. The data demonstrate successful incorporation of a neurotransmitter analog as part of a layered, silicon-based structure possessing robust and specific biomolecular composition. These findings represent a step toward the design of platforms for applications involving control and modulation of neural signaling.

Avidin↗

PubMed2004

Poly(ethylene glycol) interfaces: an approach for enhanced performance of microfluidic systems.

Microfluidic systems are extensively used platform for analytical and therapeutic applications. One of the major problems encountered in these systems is the loss of material due to non-specific surface interactions. When biological solutions are flowed through microchannels, they tend to adsorb on the surface due to the negative charge of the surface. This results in a reduced efficiency of the system which can be critical in sensitive biological analysis. Poly(ethylene glycol) (PEG) is known to form non-fouling interfaces on silicon and glass which are common materials used in microfluidic systems. The most common approach for modifying silicon/glass with PEG involves a solution phase protocol. Since the micro/nanofluidic systems have channel sizes ranging from hundreds of microns to submicron with variety of complicated network, this surface modification approach is not sufficient in forming uniform, conformal, and ultrathin films on the surface. Due to the enclosed features in these systems, the properties of liquids such as viscosity and surface tension play an important role in the clogging and eventually biofouling of these microchannels. Hence, we have developed a solvent-free vapor deposition protocol for modifying silicon/glass surfaces with PEG. Various concentrations of protein solutions were flowed through unmodified and PEG-modified glass microcapillaries of different lengths at different flow rates. PEG surfaces formed on silicon have shown 80% reduction in protein adsorption in static conditions.

Boranes↗