Punit Kohli

The emerging field of nanotube biotechnology

Nanoparticles are being developed for a host of biomedical and biotechnological applications, including drug delivery, enzyme immobilization and DNA transfection. Spherical nanoparticles are typically used for such applications, which reflects the fact that spheres are easier to make than other shapes. Micro- and nanotubes — structures that resemble tiny drinking straws — are alternatives that might offer advantages over spherical nanoparticles for some applications. This article discusses four approaches for making micro- and nanotubes, and reviews the current status of efforts to develop biomedical and biotechnological applications of these tubular structures.

Carbon Nanotube−MoS2 Composites as Solid Lubricants

Solid lubricants (SLs) characterized by low coefficients of friction (mu) and wear rates (w) drastically improve the life span of instruments that undergo extreme frictional wear. However, the performance of SLs such as sputtered or nanoparticulate molybdenum disulfide (MoS(2)), tungsten disulfide (WS(2)), or graphite deteriorates heavily under extreme operational conditions such as elevated temperatures and high humidity. Here, we present our preliminary results, which demonstrate that composites of carbon nanotubes (CNTs) and MoS(2) produced by electrodeposition of MoS(2) on vertically aligned CNT films have low mu ( approximately 0.03) and w (approximately 10(-13) mm(3)/N.mm) even at 300 degrees C, which are about 2 orders of magnitude better than those of nanoparticulate MoS(2)-based coatings. The high load-bearing capacity of CNTs provides a strong enduring support to MoS(2) nanoclusters and is responsible for their ultralow w. The incorporation of these composites in liquid lubricants reduces the friction coefficient of the liquid lubricants by approximately 15%. The technique described here to produce SL coatings with extremely appealing frictional properties will provide valuable solutions for a variety of tribological applications where the coatings encounter high temperature, reduced pressure, and/or low- and high-humidity conditions.

Investigating Photoinduced Charge Transfer in Carbon Nanotube−Perylene−Quantum Dot Hybrid Nanocomposites

In this study, we investigate photophysical and photoinduced current responses of a nanocomposite which consists of multiwalled carbon nanotubes (CNTs), thiol derivative perylene compound (ETPTCDI), and cadmium selenide quantum dots (QDs). These QDs as well as the ETPTCDI harvest photons and transfer their excited electrons or holes to CNTs to complete the circuit. Both QDs and ETPTCDI contribute charges to the carbon nanotubes, which increased the overall photon harvest efficiency of the nanocomposite. Herein, we investigate through a series of photophysical photoluminescence quenching studies the charge transfer between donors (QDs and ETPTCDI) and acceptor (CNTs). The incorporation of ETPTCDI into the nanocomposite significantly increases the adhesion between QDs and CNTs through bonding between QDs and thiol groups on ETPTCDI and π-π interactions between ETPTCDI and CNTs. Thus, ETPTCDI acted as a molecular linker between QDs and CNTs. Furthermore, a significant increase (>5 times) in the Stern-Volmer constant, K(sv), for QD emission after addition of ETPTCDI-tagged CNTs clearly indicates a large enhancement in the adhesion between CNTs and QDs. The nanocomposite shows a ∼2-4-fold increase in the photoconductivity when exposed to AM1.5 solar-simulated light. The damage to the nanocomposite from the intensity of the solar-simulated light is also investigated. The proposed nanocomposite has the potential for photovoltaic applications such as being the active component in a hybrid bulk heterojunction solar cell.

Electric field-induced direct delivery of proteins by a nanofountain probe

We report nanofabrication of protein dot and line patterns using a nanofountain atomic force microscopy probe (NFP). Biomolecules are continuously fed in solution through an integrated microfluidic system, and deposited directly onto a substrate. Deposition is controlled by application of an electric potential of appropriate sign and magnitude between the probe reservoir and substrate. Submicron dot and line molecular patterns were generated with resolution that depended on the magnitude of the applied voltage, dwell time, and writing speed. By using an energetic argument and a Kelvin condensation model, the quasi-equilibrium liquid–air interface at the probe tip was determined. The analysis revealed the origin of the need for electric fields in achieving protein transport to the substrate and confirmed experimental observations suggesting that pattern resolution is controlled by tip sharpness and not overall probe aperture. As such, the NFP combines the high-resolution of dip-pen nanolithography with the efficient continuous liquid feeding of micropipettes while allowing scalability to 1- and 2D probe arrays for high throughput.

Template-synthesized nanotubes for biotechnology and biomedical applications

Spherically shaped micro- and nano-particles are typically used for drug deliver, but this only reflects the fact that spheres are easier to make than other shapes. Micro and nanotubes, structures that resemble tiny drinking straws, are alternatives to spherical nanoparticles. Examples include organosilicon polymer nanotubes, self-assembling lipid microtubes, fullerene carbon nanotubes, template-synthesized nanotubes, and peptide nanotubes. Nanotubes offer some interesting advantages relative to spherical nanoparticles for biotechnological applications. For example, nanotubes have large inner volumes (relative to the dimensions of the tube), which can be filled with any desired chemical or biochemical species ranging in size from proteins to small molecules. In addition, nanotubes have distinct inner and outer surfaces, which can be differentially chemically or biochemically functionalized. This creates the possibility, for example, of loading the inside of a nanotube with a particular biochemical payload but imparting chemical features to the outer surface that render it biocompatible. Finally, nanotubes have open mouths, which makes the inner surface accessible and incorporation of species within the tubes particularly easy. We review here “template synthesis” method for preparing micro and nanotubes, and we give examples of recent biotech applications of these tubular systems. This review is not intended to be all encompassing; instead it discusses recent trends and emphasizes “template synthesis” method.

Advanced thermal interface materials for enhanced flip chip BGA

A family of advanced thermal interface materials for high-power flip-chip BGA (FCBGA) packages is discussed. These silver-filled adhesives provide for high reliability on laminate FCBGA packages. Laser flash thermal testing is utilized to demonstrate that these materials not only have low bulk thermal resistance, but also very low interfacial, or contact resistance. These adhesives have a low modulus and high adhesion, which enables them to flex and remain bonded as the laminate package undergoes stress during temperature cycling. Data also show that this family of TIMs has very low moisture absorption, which contributes to excellent adhesive reliability during HAST (highly accelerated stress test). Reliability data on laminate test packages will be presented. This family of materials has been shown to pass more than 1000 cycles of temperature cycling B (-55 to +125/spl deg/C, liquid-to-liquid) and 200 hours HAST (121/spl deg/C, 100% relative humidity), after JEDEC Level 3 preconditioning and three reflow simulations at 220/spl deg/C on a laminate package with 10/spl times/10 mm die. Total thermal resistance of less than 0.10 cm/sup 2/ K/W has been achieved, at a 25 /spl mu/m bondline thickness.

Investigating Ligand–Receptor Interactions at Bilayer Surface Using Electronic Absorption Spectroscopy and Fluorescence Resonance Energy Transfer

We investigate interactions between receptors and ligands at bilayer surface of polydiacetylene (PDA) liposomal nanoparticles using changes in electronic absorption spectroscopy and fluorescence resonance energy transfer (FRET). We study the effect of mode of linkage (covalent versus noncovalent) between the receptor and liposome bilayer. We also examine the effect of size-dependent interactions between liposome and analyte through electronic absorption and FRET responses. Glucose (receptor) molecules were either covalently or noncovalently attached at the bilayer of nanoparticles, and they provided selectivity for molecular interactions between glucose and glycoprotein ligands of E. coli. These interactions induced stress on conjugated PDA chain which resulted in changes (blue to red) in the absorption spectrum of PDA. The changes in electronic absorbance also led to changes in FRET efficiency between conjugated PDA chains (acceptor) and fluorophores (Sulphorhodamine-101) (donor) attached to the bilayer surface. Interestingly, we did not find significant differences in UV-vis and FRET responses for covalently and noncovalently bound glucose to liposomes following their interactions with E. coli. We attributed these results to close proximity of glucose receptor molecules to the liposome bilayer surface such that induced stress were similar in both the cases. We also found that PDA emission from direct excitation mechanism was ~2-10 times larger than that of the FRET-based response. These differences in emission signals were attributed to three major reasons: nonspecific interactions between E. coli and liposomes, size differences between analyte and liposomes, and a much higher PDA concentration with respect to sulforhodamine (SR-101). We have proposed a model to explain our experimental observations. Our fundamental studies reported here will help in enhancing our knowledge regarding interactions involved between soft particles at molecular levels.

Polydiacetylene nanovesicles as carriers of natural phenylpropanoids for creating antimicrobial food-contact surfaces.

The ultimate goal of this study was developing antimicrobial food-contact materials based on natural phenolic compounds using nanotechnological approaches. Among the methyl-β-cyclodextrin-encapsulated phenolics tested, curcumin showed by far the highest activity toward Escherichia coli with a minimum inhibitory concentration of 0.4 mM. Curcumin was enclosed in liposome-type polydiacetylene/phosholipid nanovesicles supplemented with N-hydroxysuccinimide and glucose. The fluorescence spectrum of the nanovesicles suggested that curcumin was located in their bilayer region. Free-suspended nanovesicles tended to bind to the bacterial surface and demonstrated bactericidal activity toward Gram-negative (E. coli) and vegetative cells of Gram-positive (Bacillus cereus) bacteria reducing their counts from 5 log CFU mL(-1) to an undetectable level within 8 h. The nanovesicles were covalently bound to silanized glass. Incubation of E. coli and B. cereus with nanovesicle-coated glass resulted in a 2.5 log reduction in their counts. After optimization this approach can be used for controlling microbial growth, cross-contamination, and biofilm formation on food-contacting surfaces.

Fluorescence Resonance Energy Transfer in Polydiacetylene Liposomes

Conjugated polydiacetylene (PDA) possessing stimuli-responsive properties has been intensively investigated for developing efficient sensors. We report here fluorescence resonance energy transfer (FRET) in liposomes synthesized using different molar ratios of dansyl-tagged diacetylene and diacetylene-carboxylic acid monomers. Photopolymerization of diacetylene resulted in cross-linked PDA liposomes. We used steady-state electronic absorption, emission, and fluorescence anisotropy (FA) analysis to characterize the thermal-induced FRET between dansyl fluorophores (donor) and PDA (acceptor). We found that the monomer ratio of acceptor to donor ( R ad) and length of linkers (functional part that connects dansyl fluorophores to the diacetylene group in the monomer) strongly affected FRET. For R ad = 10 000, the acceptor emission intensity was amplified by more than 18 times when the liposome solution was heated from 298 to 338 K. A decrease in R ad resulted in diminished acceptor emission amplification. This was primarily attributed to lower FRET efficiency between donors and acceptors and a higher background signal. We also found that the FRET amplification of PDA emissions after heating the solution was much higher when dansyl was linked to diacetylene through longer and flexible linkers than through shorter linkers. We attributed this to insertion of dansyl in the bilayer of the liposomes, which led to an increased dansyl quantum yield and a higher interaction of multiple acceptors with limited available donors. This was not the case for shorter and more rigid linkers where PDA amplification was much smaller. The present studies aim at enhancing our understanding of FRET between fluorophores and PDA-based conjugated liposomes. Furthermore, receptor tagged onto PDA liposomes can interact with ligands present on proteins, enzymes, and cells, which will produce emission sensing signal. Therefore, using the present approach, there exist opportunities for designing FRET-based highly sensitive and selective chemical and biochemical sensors.

Investigating surface topology and cyclic-RGD peptide functionalization on vascular endothelialization.

The advantages of endothelialization of a stent surface in comparison with the bare metal and drug-eluting stents used today include reduced late-stent restenosis and in-stent thrombosis. In this article, we study the effect of surface topology and functionalization of tantalum (Ta) with cyclic-(arginine-glycine-aspartic acid-d-phenylalanine-lysine) (cRGDfK) on the attachment, spreading, and growth of vascular endothelial cells. Self-assembled nanodimpling on Ta surfaces was performed using a one-step electropolishing technique. Next, cRGDfK was covalently bonded onto the surface using silane chemistry. Our results suggest that nanotexturing alone was sufficient to enhance cell spreading, but the combination of a nanodimpled surfaces along with the cRGDfK peptide may produce a better endothelialization coating on the surface in terms of higher cell density, better cell spreading, and more cell-cell interactions, when compared to using cRGDfK peptide functionalization alone or nanotexturing alone. We believe that future research should look into how to implement both modifications (topographic and chemical modifications) to optimize the stent surface for endothelialization.