Selective Gas-Phase Functionalization of SiO2 and SiNx Surfaces with Hydrocarbons.

Selective functionalization of dielectric surfaces is required for area-selective atomic layer deposition and etching. We have identified precursors for the selective gas-phase functionalization of plasma-deposited SiO2 and SiNx surfaces with hydrocarbons. The corresponding reaction mechanism of the precursor molecules with the two surfaces was studied using in situ surface infrared spectroscopy. We show that at a substrate temperature of 70 °C, cyclic azasilanes preferentially react with an -OH-terminated SiO2 surface over a -NHx-terminated SiNx surface with an attachment selectivity of ∼5.4, which is limited by the partial oxidation of the SiNx surface. The cyclic azasilane undergoes a ring-opening reaction where the Si-N bond cleaves upon the reaction with surface -OH groups forming a Si-O-Si linkage. After ring opening, the backbone of the grafted hydrocarbon is terminated with a secondary amine, -NHCH3, which can react with water to form an -OH-terminated surface and release CH3NH2 as the product. The surface coverage of the grafted cyclic azasilane is calculated as ∼3.3 × 1014 cm-2, assuming that each reacted -OH group contributes to one hydrocarbon linkage. For selective attachment to SiNx over SiO2 surfaces, we determined the reaction selectivity of aldehydes. We demonstrate that aldehydes selectively attach to SiNx over SiO2 surfaces, and for the specific branched aliphatic aldehyde used in this work, almost no reaction was detected with the SiO2 surface. A fraction of the aldehyde molecules reacts with surface -NH2 groups to form an imine (Si-N═C) surface linker with H2O released as the byproduct. The other fraction of the aldehydes also reacts with surface -NH2 groups but do not undergo the water-elimination step and remains attached to the surface as an aminoalcohol (Si-NH-COH-). The surface coverage of the grafted aldehyde is calculated as ∼9.8 × 1014 cm-2 using a known infrared absorbance cross-section for the -C(CH3)3 groups.

Gas Phase Organic Functionalization of SiO2 with Propanoyl Chloride.

The reaction mechanism of propanoyl chloride (C2H5COCl) with -SiOH-terminated SiO2 films was studied using in situ surface infrared spectroscopy. We show that this surface functionalization reaction is temperature dependent. At 230 °C, C2H5COCl reacts with isolated surface -SiOH groups to form the expected ester linkage. Surprisingly, as the temperature is lowered to 70 °C, the ketone groups are transformed into the enol tautomer, but if the temperature is increased back to the starting exposure temperature of 230 °C, the ketone tautomer is not recovered, indicating that the enol form is thermally stable over a wide range of temperatures. Further, the enol form is directly formed after exposure of a SiO2 surface to C2H5COCl at 70 °C. We speculate that the enol form, which is energetically unfavorable, is stabilized because of hydrogen bonding with adjacent enol groups or through hydrogen bonding with unreacted surface -SiOH groups. The surface coverage of hydrocarbon molecules is calculated as ∼6 × 1012 cm-2, assuming each reacted -SiOH group contributes to one hydrocarbon linkage on the surface. At a substrate temperature of 70 °C, the enol form is unreactive with H2O, and H2O molecules simply physisorb on the surface. At higher temperatures, H2O converts the ketone to the enol tautomer and reacts with Si-O-Si bridges, forming more -SiOH reactive sites. The overall hydrocarbon coverage on the surface can then be further increased through cycling H2O and C2H5COCl doses.

Surface Phenomena During Plasma-Assisted Atomic Layer Etching of SiO2.

Surface phenomena during atomic layer etching (ALE) of SiO2 were studied during sequential half-cycles of plasma-assisted fluorocarbon (CFx) film deposition and Ar plasma activation of the CFx film using in situ surface infrared spectroscopy and ellipsometry. Infrared spectra of the surface after the CFx deposition half-cycle from a C4F8/Ar plasma show that an atomically thin mixing layer is formed between the deposited CFx layer and the underlying SiO2 film. Etching during the Ar plasma cycle is activated by Ar+ bombardment of the CFx layer, which results in the simultaneous removal of surface CFx and the underlying SiO2 film. The interfacial mixing layer in ALE is atomically thin due to the low ion energy during CFx deposition, which combined with an ultrathin CFx layer ensures an etch rate of a few monolayers per cycle. In situ ellipsometry shows that for a ∼4 Šthick CFx film, ∼3-4 Šof SiO2 was etched per cycle. However, during the Ar plasma half-cycle, etching proceeds beyond complete removal of the surface CFx layer as F-containing radicals are slowly released into the plasma from the reactor walls. Buildup of CFx on reactor walls leads to a gradual increase in the etch per cycle.

Identification of a lysosomal pathway that modulates glucocorticoid signaling and the inflammatory response.

The antimalaria drug chloroquine has been used as an anti-inflammatory agent for treating systemic lupus erythematosus and rheumatoid arthritis. We report that chloroquine promoted the transrepression of proinflammatory cytokines by the glucocorticoid receptor (GR). In a mouse collagen-induced arthritis model, chloroquine enhanced the therapeutic effects of glucocorticoid treatment. By inhibiting lysosome function, chloroquine synergistically activated glucocorticoid signaling. Lysosomal inhibition by either bafilomycin A1 (an inhibitor of the vacuolar adenosine triphosphatase) or knockdown of transcription factor EB (TFEB, a master activator of lysosomal biogenesis) mimicked the effects of chloroquine. The abundance of the GR, as well as that of the androgen receptor and estrogen receptor, correlated with changes in lysosomal biogenesis. Thus, we showed that glucocorticoid signaling is regulated by lysosomes, which provides a mechanistic basis for treating inflammation and autoimmune diseases with a combination of glucocorticoids and lysosomal inhibitors.

Reduction of protein adsorption and macrophage and astrocyte adhesion on ventricular catheters by polyethylene glycol and N-acetyl-L-cysteine.

Cellular obstruction of poly(dimethyl)siloxane (PDMS) catheters is one of the most prevalent causes of shunt failure in the treatment of hydrocephalus. By modifying PDMS using short- and long-chain mono-functional polyethylene glycol (PEG604 and PEG5K, respectively) and N-acetyl-L-cysteine via adsorption and covalent binding (NAC and NAC/EDC/NHS, respectively), we increased surface wettability. We hypothesized that these surface modifications would inhibit protein adsorption and decrease host macrophage and astrocyte adhesion. Tested in a bioreactor set to mimic physiological flow, all modified surfaces significantly decreased albumin adsorption compared with PDMS (p < 0.05) except for PEG604-modified PDMS (p = 0.14). All four modification strategies significantly reduced (p < 0.01) fibronectin adsorption. PEG604, PEG5K, NAC, and NAC/EDC/NHS reduced the average level of macrophage adhesion by 53%, 63%, 40%, and 58% (p <.0.05 except when comparing PDMS with NAC) and astrocyte adhesion by 47%, 83%, 91%, and 72% (p < 0.05 except when comparing PDMS with PEG604), respectively. Combined with saline soak results which suggest that the surface wettability is stable over 30 days for each modification, our results are consistent with the hypothesis that these modifications decrease cell adhesion on catheters in vitro for the treatment of hydrocephalus.

Effects of surface wettability, flow, and protein concentration on macrophage and astrocyte adhesion in an in vitro model of central nervous system catheter obstruction.

While silicone devices have vastly improved an array of medical treatments, reactions at the tissue-substrate interface often impede their functionality. Insertion of a poly(dimethyl)siloxane (PDMS) catheter into the cerebral ventricles to drain excess cerebrospinal fluid (CSF) is the most common treatment of hydrocephalus, but shunting often fails because inflammatory tissue, choroid plexus cells, and debris grow into these central nervous system catheters and obstruct flow. We hypothesized that plasma oxidation of PDMS would inhibit macrophage and astrocyte adhesion under flow (0 to 0.3 mL/min) and protein (20.8 to 240 mg/dL) conditions similar to those observed in the physiological state. Oxidation (to increase wettability) had an inhibitory effect on macrophage cell binding (yielding a significant 88% change) that was generally more pronounced than the effect of flow (22% change) or protein concentration (3% change). In contrast, greater flow increased binding of astrocytes in most cases (yielding a significant 97% change); plasma oxidation (19% change), and protein concentration (60% change) had less pronounced effects. This study is the initial indicator that plasma oxidation of PDMS catheters may inhibit macrophage adhesion during CSF outflow but may not be as effective at inhibiting astrocyte binding.

Mechanical contributions to astrocyte adhesion using a novel in vitro model of catheter obstruction.

Drainage and diversion of cerebrospinal fluid (CSF) through shunt systems is the most common treatment for hydrocephalus, but complications due to tissue obstruction of the catheter occur in up to 61% of patients. Although shunt systems have undergone limited technological advancements to resist mammalian cell adhesion, there is a need to further reduce adhesion that can exacerbate obstruction. The high intrinsic variability in clinical studies and an inability to predict chronic adhesion of host cells in vitro while maintaining the environmental conditions observed in hydrocephalus have impeded progress. We designed the hydrocephalus shunt catheter bioreactor (HSCB) to measure inflammatory cell adhesion under experimentally manipulated conditions of CSF pressure, pulsation rate, and flow rates. For a 20-h period, astrocytes were perfused through the pulsatile flow system, and adhesion on silicone catheters was recorded. These results were compared with those obtained under static cell culture conditions. Astrocyte adhesion was significantly increased under conditions of increased flow rate (0.25 and 0.30 mL/min), and a trend toward increased adhesion was observed under conditions of elevated pressure and pulsation rate. Because the HSCB represents physiologic conditions more accurately than static cell culture, our results suggest that standard static cell culturing techniques are insufficient to model inflammatory cell adhesion on catheters used in the treatment of hydrocephalus and that changes to the ventricular microenvironment can alter the mechanisms of cellular adhesion. The HSCB represents a relevant test system and is an effective model system for the analysis of cellular adhesion and occlusion of shunt catheters.

Use of the ultraviolet absorption spectrum of CF2 to determine the spatially resolved absolute CF2 density, rotational temperature, and vibrational distribution in a plasma etching reactor.

Broadband ultraviolet absorption spectroscopy has been used to determine CF(2) densities in a plasma etch reactor used for industrial wafer processing, using the CF(2) A (1)B(1)<--X (1)A(1) absorption spectrum. Attempts to fit the experimental spectra using previously published Franck-Condon factors gave poor results, and values for the higher vibrational levels of the A state [(0,v(2),0), with v(2) (')>6] from the ground state were missing; hence new values were calculated. These were computed for transitions between low-lying vibrational levels of CF(2) X (1)A(1) to vibrational levels of CF(2) A (1)B(1) (v(1) ('),v(2) ('),0) up to high values of the vibrational quantum numbers using high level ab initio calculations combined with an anharmonic Franck Condon factor method. The Franck Condon factors were used to determine the absorption cross sections of CF(2) at selected wavelengths, which in turn were used to calculate number densities from the experimental spectra. Number densities of CF(2) have been determined in different regions of the plasma, including the center of the plasma and outside the plasma volume, and CF(2) rotational temperatures and vibrational energy distributions were estimated. For absorption spectra obtained outside the confined plasma volume, the CF(2) density was determined as (0.39+/-0.08)x10(13) molecule cm(-3) and the vibrational and rotational temperatures were determined as 303 and 350 K, respectively. In the center of the plasma reactor, the CF(2) density is estimated as (3.0+/-0.6)x10(13) molecules cm(-3) with T(rot) approximately 500 K. The fitted vibrational distribution in the CF(2) ground state corresponds to two Boltzmann distributions with T(vib) approximately 300 and T(vib) approximately 1000 K, indicating that CF(2) molecules are initially produced highly vibrationally excited, but are partially relaxed in the plasma by collision.

Quantifying cell scattering: the blob algorithm revisited.

BACKGROUND: A method to objectively quantify cell scattering would permit quantitative evaluation of therapies and compounds intended to affect this physiologic process, which has relevance to normal (e.g., development) and pathologic (e.g., metastasis) events. METHODS: A grid-based modified blob analysis was performed on a set of images of Madin-Darby Canine Kidney (MDCK) cells to quantify the following parameters: the number of cellular clusters in each image, the size of the clusters in terms of pixel counts, and the number of cells in each cluster. These parameters were used as measures of cell scattering and were compared with subjective assessments of scattering made by three experienced examiners. RESULTS: The quantitative parameters correlated strongly to subjective assessments. The algorithm displayed a different concept of "clustering" than the examiners and consistently identified more clusters than did the examiners. There was close agreement in the number of cells counted. All three quantitative parameters correlated strongly to the subjective scattering scores, as follows: cluster count (r(s) = -0.765 to -0.789, P < 0.0001), cluster size in pixels (r(s) = 0.838 to 0.845, P < 0.0001), and cluster size in cells (r(s) = 0.758 to 0.804, P < 0.0001). The parameters were continuous, providing greater resolving power than ordinal subjective scores. CONCLUSIONS: The findings confirmed that our algorithm reproduces the traditional classification of scattering with improved resolution, quantification, and objectivity.