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PubMed2004

Inflammatory time course after quartz instillation: role of tumor necrosis factor-alpha and particle surface.

Inflammation has been suggested as the key factor in the development of quartz-induced fibrosis and carcinogenesis, and particle surface properties are argued as an important characteristic responsible for these pathologic alterations. To evaluate the effect of surface modification on acute and subchronic inflammation, female Wistar rats were intratracheally instilled with 2 mg native quartz, or quartz coated either with polyvinyl-pyridine-N-oxide or with aluminium lactate. Various markers of lung toxicity, inflammation, and oxidative stress were found to be enhanced at 3, 7, 21, and 90 d after instillation of native quartz. Quartz-treated animals also showed enhanced immunostaining of nuclear factor-kappaB (NF-kappaB) in alveolar macrophages and lung epithelium, as well as reduced IkappaBalpha levels in whole lung homogenate. Both surface modifications were found to inhibit most of the effects as observed with native quartz. NF-kappaB activation was also observed in vitro in rat lung epithelial cells following treatment with lavage fluid from quartz-treated animals, as well as with conditioned medium of quartz-treated macrophages, and these effects appeared to be at least partly tumor necrosis factor-alpha-independent. In conclusion, the persistent subchronic inflammatory lung response after quartz exposure appears to be particle surface-driven and is associated with NF-kappaB activation in both alveolar macrophages and the lung epithelium.

Animals↗

PubMed1981

The effect of surface membrane modifications of fibroblastic cells on the entry process of Trypanosoma cruzi trypomastigotes.

Treatment prior to infection with Trypanosoma cruzi trypomastigotes of Vero, MA-103, and chick muscle cells with concanavalin A, phytohemagglutinin, wheat germ agglutinin and ricin I results in a diminished parasite interiorization in these cells; succinylated concanavalin A is also inhibitory. The effect of these lectins is abolished by the corresponding sugar haptens. Trypsin and periodate treatment of the cells also inhibits infection, as well as calcium ionophore A23187 and drugs that disrupt microtubules and microfilaments directly, like colchicine, vinblastine and cytochalasin B. These results show that alteration(s) of a surface glycoprotein(s) and/or of the plasma membrane architecture of fibroblastic host cells inhibit infection, suggesting that the surface membrane of these cells does not play a passive role in the process of infection by T. cruzi.

Agglutinins↗

PubMed2003

Surface functionalization of polypyrrole film with glucose oxidase and viologen.

A surface modification technique was developed for the functionalization of polypyrrole (PPY) film with glucose oxidase (GOD) and viologen moieties. The PPY film was first graft copolymerized with acrylic acid (AAc) and GOD was then covalently immobilized through the amide linkage formation between the amino groups of the GOD and the carboxyl groups of the grafted AAc polymer chains in the presence of a water-soluble carbodiimide. Viologen moieties could also be attached to the PPY film via graft-copolymerization of vinyl benzyl chloride with the PPY film surface followed by reaction with 4,4'-bipyridine and alpha,alpha'-dichloro-p-xylene. X-ray photoelectron spectroscopy (XPS) was used to characterize the PPY films after each surface modification step. Increasing the AAc graft concentration would allow a greater amount of GOD to be immobilized but this would decrease the electrical conductivity of the PPY film. The activity of the immobilized GOD was compared with that of free GOD and the kinetic effects were also studied. The immobilized GOD was found to be less sensitive to temperature deactivation as compared to the free GOD. The results showed that the covalent immobilization technique offers advantages over the technique involving the entrapment of GOD in PPY films during electropolymerization. The presence of viologen in the vicinity of the immobilized GOD also enabled the GOD-catalyzed oxidation of glucose to proceed under UV irradiation in the absence of O(2).

Acrylates↗

PubMed2001

Dental hard tissue modification and removal using sealed transverse excited atmospheric-pressure lasers operating at lambda=9.6 and 10.6 microm.

Pulsed CO(2) lasers have been shown to be effective for both removal and modification of dental hard tissue for the treatment of dental caries. In this study, sealed transverse excited atmospheric pressure (TEA) laser systems optimally tuned to the highly absorbed 9.6 microm wavelength were investigated for application on dental hard tissue. Conventional TEA lasers produce an initial high energy spike at the beginning of the laser pulse of submicrosecond duration followed by a long tail of about 1-4 micros. The pulse duration is well matched to the 1-2 micros thermal relaxation time of the deposited laser energy at 9.6 microm and effectively heats the enamel to the temperatures required for surface modification at absorbed fluences of less than 0.5 J/cm(2). Thus, the heat deposition in the tooth and the corresponding risk of pulpal necrosis from excessive heat accumulation is minimized. At higher fluences, the high peak power of the laser pulse rapidly initiates a plasma that markedly reduces the ablation rate and efficiency, severely limiting applicability for hard tissue ablation. By lengthening the laser pulse to reduce the energy distributed in the initial high energy spike, the plasma threshold can be raised sufficiently to increase the ablation rate by an order of magnitude. This results in a practical and efficient CO(2) laser system for caries ablation and surface modification.

Dental Caries↗

PubMed2004

Supported cell mimetic monolayers and their interaction with blood.

Surface modification using supported monolayers of phosphorylcholine containing phospholipids has been an accepted strategy for developing blood-contacting materials. We present a detailed study of the blood compatibility of the supported monolayers of phospholipid, glycolipid, and cholesterol (Chol) binary and ternary lipid combinations using in vitro techniques. The packing and orientation of these monolayers have been correlated with the blood compatibility. We have used phosphatidylcholine (PTC) for phospholipid, galactocerebroside (Gal) for glycolipid, and Chol based on the headgroup structure to represent the major lipid components of the endothelial luminal cell membrane. The interfacial behavior of various combinations of PTC, Gal, and Chol monolayers have been studied at the air/water interface and deposited on hydrophobic polycarbonate (PC) polymer substrates with the help of the Langmuir-Blodgett trough. The packing and orientation of the supported monolayers have been varied by means of changing the lipid composition rather than the deposition parameters. This approach seems to be more similar to the in vivo conditions. The different supported monolayer surfaces prepared accordingly are (1) a closely packed ordered hydrophobic surface, PC modified with the combination PTC/Chol/Gal (1:0.35:0.125), (2) a loosely packed ordered hydrophobic surface, PC modified with the combination PTC/Chol (1:0.35), and (3) a closely packed ordered hydrophilic surface, PC modified with the combination PTC/Chol (1:0.7). An optimized modified surface (PTC/Chol/Gal, 1:0.35:0.125) has been identified on the basis of the maximum transfer ratio from the air/water interface and characterized by using atomic force microscopy. The concentration of Chol has been found to be an important parameter, which influences the transfer ratio. The Gal improves the monolayer integrity under a reduced Chol concentration. The blood compatibility of these supported monolayers was studied by protein adsorption, blood cell adhesion, and calcification. The tightly packed ordered hydrophobic surface (PTC/Chol/Gal, 1:0.35:0.125), has been found to be more blood compatible because of reduced blood cell adhesion and calcification. This surface also promotes albumin adsorption and may be the reason for the reduced platelet activation, while in the case of the loosely packed ordered hydrophobic surface (PTC/Chol, 1:0.35) the protein adsorption also has been reduced along with the blood cell adhesion and calcification. When the ordered hydrophilic surface (PTC/Chol, 1: 0.7) of the monolayer has been exposed, the blood cell adhesions as well as the overall protein adsorption were significantly reduced. However, the packing of the phosphorylcholine moieties of the polar headgroup has been affecting the calcification on the surface. We have observed an increase in calcification to the surface modified with the loosely packed polar headgroup, from a relative study on chitosan and chitosan modified with the monolayer of PTC. These findings are helpful for the surface modifications for blood-contacting materials using this strategy.

Air↗

PubMed2002

Interventions for replacing missing teeth: different types of dental implants.

BACKGROUND: Dental implants are available in different materials, shapes and with different surface characteristics. In particular, numerous implant surface modifications have been developed for enhancing clinical performances. OBJECTIVES: To test the null hypothesis of no difference in clinical performance between various root-formed osseointegrated implant types. SEARCH STRATEGY: The Cochrane Oral Health Group Specialised Trials Register, The Cochrane Controlled Trials Register, MEDLINE and EMBASE were searched. Hand searching included several dental journals. Bibliographies of relevant clinical trials and review articles were checked for studies outside the handsearched journals. In addition, authors of all identified trials and fifty-five oral implant manufacturers were contacted to find unpublished or ongoing RCTs. Two extensive personal libraries (ME and AJ) were consulted. The last electronic search was conducted 8th May 2002. SELECTION CRITERIA: All randomised controlled trials of oral implants comparing implants with different materials, shapes and surface properties having a follow-up of at least one year. DATA COLLECTION AND ANALYSIS: Data were independently extracted, in duplicate, by two reviewers (ME & HW). Authors were contacted for details of randomisation and withdrawals and a quality assessment was carried out. The Cochrane Oral Health Group's statistical guidelines were followed. MAIN RESULTS: Thirty publications, representing 13 different RCTs, were identified. Five of these RCTs (seven publications), which reported results from a total of 326 patients, were suitable for inclusion in the review. Six implant systems were compared: Astra, Branemark, IMZ, ITI, Steri-Oss and Southern with a follow-up ranging from one to three years. All implants were made in commercially pure titanium and had different shapes and surface preparations. On a patient rather than per implant basis there were no statistically significant differences for failures and marginal bone level changes on intra-oral radiographs between various implant systems. REVIEWER'S CONCLUSIONS: There was no evidence that any of the implant systems evaluated was superior to the other. However, these findings are based on a few RCTs all having short follow-up periods and few participants. More RCTs should be conducted, with follow-up of at least five years and including a sufficient number of patients, to detect a true difference if any exists. Such trials should be reported according to the CONSORT guidelines (http://www.consort-statement.org/).

Dental Implantation, Endosseous↗

PubMed2001

Adhesion of corneal epithelial cells to cell adhesion peptide modified pHEMA surfaces.

Epithelialization of a corneal implant is a desirable property. In this study we compared surface modification of poly (2-hydroxyethyl methacrylate) (pHEMA) with the cell adhesion peptides RGDS and YIGSR. Various parameters in the tresyl chloride activation and modification reactions were considered in order to maximize surface coverage with the peptide including tresyl chloride reaction solvent. tresyl chloride reaction time, tresyl chloride concentration, peptide concentration, and peptide reaction pH. Surface chemistry and corneal epithelial cell adhesion to the modified surfaces were examined. X-ray photoelectron spectroscopy data suggested that while peptide modification had occurred, surface coverage with the peptide was incomplete. Acetone was found to result in a higher fraction of nitrogen and surface bound carboxyl groups compared to dioxane and ether. Furthermore, corneal epithelial cell adhesion to the surfaces for which acetone was used for the activation reaction was significantly greater. Statistical analysis of the various samples suggests that lower peptide concentrations and higher tresyl chloride reaction times result in better cell adhesion. Furthermore, modification with YIGSR resulted in higher surface concentrations and better cell adhesion than modification with RGDS. Little or no cell adhesion was noted on the unmodified pHEMA controls. Protein adsorption results suggest that the differences in cell adhesion cannot be attributed to differences in serum protein adsorption from the culture medium. We conclude that YIGSR modified surfaces have significant potential for further development in corneal applications.

Adsorption↗

PubMed2006

The influence of surface bioactivated modification on titanium percutaneous implants anchored in bone.

In order to achieve biological sealing and resist mechanical damage of load-bearing percutaneous devices, Ti with excellent mechanical properties was anodic-oxidized to be endowed with bioactivity, with plasma-sprayed hydroxyapatite coated Ti as control. Similar to previous works, hydroxyapatite coating could bond tightly with living tissues, resulting in implant stability for whole implantation periods. Meanwhile, when anodic-oxidized bioactivated Ti was implanted percutaneously in vivo, it could induce a layer of calcium phosphate at the interface of tissues/implant. This layer of Ca-P not only induced the fibrous tissue or collagen ingrowth in its structure, but also improved the osteointegration between the bone and the implant. There was no significant biological response difference for the anodic-oxidized Ti and HA coated Ti at different implantation period with histological statistical analysis (p>0.05). Accordingly, suitable bioactivated modified surface of Ti by anodic-oxidized method could not only obtain the same results as the HA coating, but also might avoid some drawbacks of plasma-sprayed HA coatings to achieve biological sealing for a long period in vivo.

Animals↗

PubMed2006

Effect of quaternary ammonium cations on dye sorption to fly ash from aqueous media.

The efficacy of the surface modification of fly ash by quarternary ammonium cations in the removal of dyes from aqueous solution is demonstrated. A series of organo-fly ash materials were synthesized by treating fly ash with quarternary ammonium cations such as tetraethylammonium, hexadecyltrimethylammonium, and benzyldimethyltetradecylammonium (TEA, HDTMA, and BDTDA). Two types of dyes were used for the investigation, disperse and anionic dyes. The effects of initial dye concentration, contact time, temperature, and the mechanism of dye sorption were investigated. The sorption was found to be affected by the structure and size of the quaternary ammonium cations as well as that of the dyes. Sorption of dyes was considerably enhanced by the surface modification. Thermodynamic parameters such as free energy (DeltaG0), enthalpy (DeltaH0), and entropy (DeltaS0) for the sorption process were also calculated.

Journal Article↗

PubMed2006

Spectroelectrochemical investigation of a flavoprotein with a flavin-modified gold electrode.

A flavin-modified gold electrode was developed in order to catalyze the electrochemical oxidoreduction of flavoproteins. Surface modification was carried out by a two-step procedure. In the first step a mixed self-assembled monolayer obtained by adsorption of activated and nonactivated 3,3'-dithiopropionic acid (free acid and N-succinimidyl ester) was formed, followed by the covalent attachment of a N(10)-hexylamino-alkylated flavin derivative via an amide bond in the second step. The electrochemical properties of the flavin-modified electrode are presented and discussed. The redox potential of the attached flavin was measured at various pH values and the electron-transfer rate constant between electrode and flavin was determined as k0 = 5 s(-1) independent of pH. The flavin-modified electrode was successfully applied to the electrochemical and spectroelectrochemical investigation of the flavoprotein WrbA from Escherichia coli that shows some structural similarities to flavodoxins. It is concluded that the electron transfer "electrode --> flavin --> flavoprotein" occurs by a two-step hopping mechanism where the first step is rate determining. Kinetic details are discussed. Furthermore, it turned out that, in contrast to flavodoxins, where the semiquinone state is stabilized, WrbA rapidly takes up two electrons, directly leading to the fully reduced form. The presented electrode surface modification may generally lend itself for spectroelectrochemical investigations of flavoproteins.

Adsorption↗

PubMed2004

In vitro analysis of modified surfaces of silicone breast implants.

BACKGROUND: Although silicone breast implants are well tolerated, local complications such as capsular contracture occur because of insufficient integration with surrounding tissues. In this study, cell behaviour on hydrophilized silicone breast implant foils was analysed qualitatively and quantitatively under in vitro conditions in order to provoke the desired responses in a defined environment. METHODS: Silicone breast implant foils with different surface modifications were tested after 24 hours, 5 days and 7 days. The following modifications of silicone implant foils were tested: Unmodified silicone, silicone after-graft polymerisation for polyacrylic acid (pAAc), silicone-pAAc-fibronectin adsorptive, silicone-pAAC-fibronectin covalent, positive and negative controls. Experiments were conducted using cell culture with murine mouse fibroblasts L-929. Cytotoxicity assays were carried out in direct and indirect contact with cells grown on the material. For the viability test and qualitative analysis of cell proliferation on different foils, both fluoresceine-diacetate and ethidiumbromide were used and in addition the morphologic description of hemalaun-stained cells were used. Quantitative cell analysis was carried out using XTT after resuspension. RESULTS: Toxic influence on cell cultures could be excluded for coated and uncoated surfaces in contact with dissolved biomaterials. Unmodified silicone surfaces showed poor cell growth in direct contact. We found a gradual improvement of cell morphology, with the spread and proliferation depending on the type of surface modification. Better results were achieved with covalently coupled fibronectin and GRGDS than with pAAc. CONCLUSION: Covalent immobilisation of hydrophobic silicone rubber can improve the initial cell-biomaterial interactions that are required to aid the successful development of tissue-like structures.

Biocompatible Materials↗

PubMed1998

A critique of the evidence for scavenging by Neanderthals and early modern humans: new data from Kobeh Cave (Zagros Mountains, Iran) and Die Kelders Cave 1 layer 10 (South Africa).

The primary mode of faunal exploitation by Neandertals and early modern humans remains a debated topic. Binford (1981, 1984, 1985, 1988) has argued for an obligate scavenging mode, Stiner (1991a, 1991b, 1991c, 1993, 1994) for a more opportunistic scavenging mode, while other researchers (Chase, 1986, 1988, 1989; Klein, 1989, 1994, 1995; Klein & Cruz-Uribe, 1996) deny the importance of scavenging as a faunal exploitation tactic. The scavenging interpretations rely primarily on several patterns in the faunal remains: the presence of a skeletal element pattern dominated by heads or head and foot parts, the presence of carnivore tooth marks on bone fragments, and infrequent cut marks that typically are not located on shaft regions of long bones or on fleshy bones. Five sites have been used to argue for scavenging: Klasies River Mouth, Combe Grenal, Grotta Guattari, Grotta dei Moscerini, and Grotte Vaufrey. The former four of the five sites are biased samples in that long bone shafts and other difficult to identify fragments were discarded at excavation. The analysis of Grotte Vaufrey included only those shafts identifiable to species or genus, thus excluding the vast majority of shaft specimens. This bias systematically shapes the skeletal element and surface modification patterning in ways that make the assemblages appear to fit a model of scavenging, when in fact the main determinant of the pattern is the bias in the flawed samples. This problem is illustrated with two unbiased faunal assemblages (Kobeh Cave and Die Kelders Layer 10). Skeletal element abundance is calculated in a way that mimics the bias in the sites listed above by excluding the shafts. Using this procedure, both Kobeh and Die Kelders have a head and foot skeletal element pattern and thus appear scavenged. Both assemblages are then analyzed in their entirety and a new pattern, consistent with hunting, is revealed. Taphonomic data on bone survival and destruction provide an explanation for this result. Excluding shaft fragments from the analysis also biases the surface modification patterning in such a way as to produce a pattern more consistent with scavenging. The conclusion is that there is no reliable evidence for scavenging by Neandertals or early modern humans.

Animals↗

PubMed1994

Blood compatibility of sputter-deposited alumina films.

The surface modification of metals by the application of blood compatible ceramics is one approach to developing durable and blood-compatible materials. The blood compatibility of sputter-deposited alumina films was investigated in vitro. The alumina films were prepared by reactive sputtering and conventional sputtering. Diffractometer studies of the alumina films indicate that the sputter-deposited alumina films have amorphous structures. Electron spectroscopy chemical analysis studies of the alumina films indicate that the sputter-deposited alumina films are nonstoichiometric (O/A1 ratio of 1.15-1.66). We examined the platelet reaction to the alumina films and the intrinsic coagulation factor XII activation by the alumina films. Medical grade segmented polyurethane was also tested. The alumina films experienced an adhesion of about 50% fewer platelets than the segmented polyurethane. Also, fewer platelet morphologic changes were observed on the alumina films than on the segmented polyurethane. Factor XII activation was less on the alumina films than on the segmented polyurethane. Surface modification by the sputter-deposited alumina films is promising for developing blood-compatible and durable materials.

Aluminum Oxide↗

PubMed2000

Porous polyethylene as a spacer graft in the treatment of lower eyelid retraction.

PURPOSE: An experimental study was performed to: 1) assess the tolerance and incorporation of porous polyethylene (Medpor) in the posterior lamella of the rabbit lower eyelid; 2) analyze the effect of implant thickness on incorporation; 3) investigate the ability of conjunctiva to grow over vascularized Medpor and; 4) determine the effects of Medpor surface modification on biocompatibility and fibrovascularization. METHODS: In phase I, 10 rabbit eyelids were operated on to analyze the effects of implant thickness and to develop the surgical technique used in phase II of the study. In phase II, 20 lower eyelids of 10 rabbits received 0.85-mm-thick Medpor grafts, each rabbit receiving both an uncoated implant and one coated with an immobilized collagen. RESULTS: There were no extrusions in phase II. with a postoperative follow-up from 14 to 17 weeks. Fourteen of 20 eyelids had full-thickness conjunctival incisions or excisions placed over the Medpor implant to determine the growth potential of conjunctiva over a vascularized implant. All but one eyelid showed complete defect coverage, occurring in as little as 3 days. Histopathology indicated complete Medpor fibrovascularization as early as 4 weeks after implantation. Because neither coated nor uncoated implants extruded in phase II, no conclusions can be drawn regarding the efficacy of Medpor surface modification. CONCLUSION: Medpor was well tolerated in this soft tissue application, and it offers advantages over other graft materials.

Animals↗

PubMed1998

In vitro blood compatibility of surface-modified polyurethanes.

Polyurethanes have proven durable materials for the manufacture of flexible trileaflet heart valves, during in vitro tests. The response of two polyurethanes of differing primary structure to parameters of blood compatibility has now been investigated, using an in vitro test cell. Platelet (beta-thromboglobulin) release, complement (C3a) activation, the activation of free plasma and surface-bound factor XII were studied using fresh, human blood (no anticoagulant) or citrated plasma in control and surface-modified polyurethane. Surface modifications were designed to affect material thrombogenicity and included covalent attachment of heparin, taurine, a platelet membrane glycoprotein fragment, polyethylene oxide (PEO), 3-aminopropyltriethoxysilane, and glucose or glucosamine. Unmodified control polyurethanes caused platelet release and complement activation. High molecular weight (2000 D) polyethylene oxide reduced platelet release slightly but only glucose attachment to the surface produced a significant reduction in platelet activation. All modifications reduced C3 activation compared with controls, but the greatest reduction was achieved with polyethylene oxide attachment or glycosylation. Most surface modifications were more activating of factor XII, both in plasma and on the material surfaces, than the control polyurethanes. Heparin and high molecular weight PEO produced the greatest activation of factor XII in the free plasma form, but low molecular weight PEO and glucosamine produced the greatest activation of surface-bound factor XIIa. The least activating surfaces, affecting both free plasma and surface-bound factor XIIa, were those treated with platelet membrane glycoprotein fragment and glucose. PEO surfaces performed relatively well, compared with controls and most surface modifications. The best overall surface, however, was the glucose-modified surface which was least activating considering all parameters of blood compatibility.

Biocompatible Materials↗

PubMed2006

Modifications of surface carbohydrates on bovine spermatozoa mediated by oviductal fluid: a flow cytometric study using lectins.

The objective of the present study was to characterize and quantify changes in exposed saccharide residues of bovine sperm during capacitation in oviductal fluid (ODF) using flow cytometry (FC). Bovine sperm were incubated with 0% or 50% non-luteal ODF for 30 min or 3.5 h. After incubation, sperm were labelled with 11 fluorescein isothiocyanate-labelled lectins and evaluated for lectin binding with FC. Furthermore, inhibiting sugars were used to determine specificity of lectin binding to oligosaccharides on the sperm surface. After 30 min incubation, there was a 91% decrease in fluorescence intensity of labelled sperm incubated in WGA, a 76% decline for Con A, 75% decline for BS-I and a 36% decline for DBA. These differences remained approximately the same over the 3.5-h incubation. Interestingly, although there was no reduction in UEA-I binding at 30 min, a significant reduction (23%) was observed at 3.5 h. Con A fluorescence was mostly inhibited with either alpha-d-glucose or alpha-d-mannose (86% and 90% respectively). BS-I fluorescence was reduced after prior incubation of the control samples with N-acetyl-galactosamine and galactose by 74% and 80% respectively. After prior incubation with N-acetyl-galactosamine DBA fluorescence reduced by 18% in the control samples. With UEA-I no fluorescence reduction was observed after prior incubation with l-fucose. We have demonstrated that capacitation of bovine sperm in ODF is accompanied by a quantitative reduction in individual lectin binding sites. These modifications may be crucial to the subsequent signalling events involved with sperm-zona binding, zona penetration or interaction with the oolema.

Animals↗

PubMed2006

Luteinizing hormone-releasing hormone agonist and transferrin functionalizations enhance nanoparticle delivery in a novel bovine ex vivo eye model.

PURPOSE: To determine whether topical ocular delivery of <100 nm nanoparticles can be enhanced by coating their exterior with peptide or protein ligands for cell surface receptors. METHODS: A novel ex vivo bovine eye model was validated for its integrity up to 60 min. Using this model, the uptake of 20 nm polystyrene nanoparticles (administered as a single 50 mul drop) before and after surface conjugation with deslorelin, a luteinizing hormone-releasing hormone (LHRH) agonist, or transferrin was determined at 5 and 60 min in individual layers of cornea and aqueous humor. Selected studies were done in the absence of corneal epithelium in the ex vivo model or using excised cornea and conjunctiva. LHRH and transferrin receptor mRNA and protein expression in corneal epithelium and conjunctiva were determined by real-time PCR and western blot, respectively. RESULTS: Corneal histology, ZO-1 immunostain pattern, and mannitol permeability were similar in controls and at the end of the ex vivo study. Corneal epithelial nanoparticle uptake in the absence of surface modification was 1.1-1.6% at 5 min and remained at about this level even at 60 min. Removal of the corneal epithelium resulted in about 22% particle uptake in the corneal stroma at 5 and 60 min compared to about 0.5% in the presence of epithelium, indicating the barrier nature of corneal epithelium. Deslorelin and transferrin conjugation enhanced corneal epithelial uptake of nanoparticles by 3- and 4.5 fold at 5 min and by 4.5- and 3.8 fold at 60 min, respectively. The total corneal uptake in 5 min is approximately 2.4, 9, and 16% with plain, deslorelin-functionalized, and transferrin-functionalized nanoparticles. In all groups, the nanoparticle uptake per unit tissue weight was in the order: corneal epithelium>stroma>endothelium with levels in the aqueous humor being undetectable. In excised cornea and conjunctiva studies, nanoparticle transport and uptake was elevated for both deslorelin and transferrin conjugated nanoparticles. Expression of LHRH and transferrin receptors was observed in corneal epithelium as well as conjunctiva. CONCLUSIONS: The ex vivo bovine eye model is a useful tool in understanding disposition of nanoparticles after topical delivery. The corneal epithelium is a significant barrier for topical nanoparticle delivery to the anterior segment. Surface modification of nanoparticles by conjugating an LHRH agonist or transferrin is a useful approach to provide rapid, efficient delivery of intact nanoparticles into and/or across cornea and conjunctiva.

Animals↗

PubMed2004

Gold-coated microelectrode array with thiol linked self-assembled monolayers for engineering neuronal cultures.

We report the use of a gold coating on microelectrode arrays (MEAs) to enable the use of the relatively reliable surface modification chemistry afforded by alkanethiol self-assembled monolayers (SAMs). The concept is simple and begins with planar MEAs, which are commercially available for neuronal cell culture and for brain slice studies. A gold film, with an intermediate adhesive layer of titanium, is deposited over the insulation of an existing MEA in a manner so as to be thin enough for transmission light microscopy as well as to avoid electrical contact to the electrodes. The alkanethiol-based linking chemistry is then applied for the desired experimental purpose. Here we show that polylysine linked to alkanethiol SAM can control the geometry of an in vitro hippocampal neuronal network grown on the MEA. Furthermore, recordings of neuronal action potentials from random and patterned networks suggest that the gold coating does not significantly alter the electrode properties. This design scheme may be useful for increasing the number of neurons located in close proximity to the electrodes. Realization of in vitro neuronal circuits on MEAs may significantly benefit basic neuroscience studies, as well as provide the insight relevant to applications such as neural prostheses or cell-based biosensors. The gold coating technique makes it possible to use the rich set of thiol-based surface modification techniques in combination with MEA recording.

Action Potentials↗