Analysis of protein adsorption characteristics to nano-pore silica particles by using confocal laser scanning microscopy

Suh, Chang Woo; Kim, Min Young; Choo, Jae Bum; Kim, Jong Kil; Kim, Ho Kun; Lee, Eun Kyu

doi:10.1016/j.jbiotec.2004.05.005

Cited by 61 publications

(34 citation statements)

References 16 publications

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“…Midy et al proved that the higher adsorption of basic fibroblast growth factor on carbonated apatite than HA could be ascribed to the higher SSA, which gave the carbonated apatite a higher surface reactivity [43]. Suh et al reported that the amount of BSA adsorbed on silicon particles increased significantly with increasing size of the micropores, and this effect was in addition to the contribution of SSA to BSA adsorption [44]. So, higher microporosity might be another determinant of greater amounts of adsorbed protein on the BCP surface.…”

Section: Discussionmentioning

confidence: 99%

Effect of phase composition and microstructure of calcium phosphate ceramic particles on protein adsorption

et al. 2010

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Section: Discussionmentioning

confidence: 99%

Effect of phase composition and microstructure of calcium phosphate ceramic particles on protein adsorption

et al. 2010

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“…Such alterations occur as a result of nanofeatures approaching protein molecular dimensions, effectively disrupting or encouraging the adsorption of proteins needed to facilitate cell adhesion (fibronectin, vitronectin, etc.). 30,31 Indeed, it has been shown experimentally that substrate topography has a direct influence on the composition, orientation, or conformation of adsorbed ECM components. 32 Second, it can be postulated that pits of this scale have an effect on the physical properties of the substrate, affecting surface-free energy, and hence, potential surface wettability.…”

Section: Discussionmentioning

confidence: 99%

Regulation of implant surface cell adhesion: characterization and quantification of S‐phase primary osteoblast adhesions on biomimetic nanoscale substrates

Biggs

Richards

Gadegaard

et al. 2006

Journal Orthopaedic Research

109

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Integration of an orthopedic prosthesis for bone repair must be associated with osseointegration and implant fixation, an ideal that can be approached via topographical modification of the implant/bone interface. It is thought that osteoblasts use cellular extensions to gather spatial information of the topographical surroundings prior to adhesion formation and cellular flattening. Focal adhesions (FAs) are dynamic structures associated with the actin cytoskeleton that form adhesion plaques of clustered integrin receptors that function in coupling the cell cytoskeleton to the extracellular matrix (ECM). FAs contain structural and signalling molecules crucial to cell adhesion and survival. To investigate the effects of ordered nanotopographies on osteoblast adhesion formation, primary human osteoblasts (HOBs) were cultured on experimental substrates possessing a defined array of nanoscale pits. Nickel shims of controlled nanopit dimension and configuration were fabricated by electron beam lithography and transferred to polycarbonate (PC) discs via injection molding. Nanopits measuring 120 nm diameter and 100 nm in depth with 300 nm center-center spacing were fabricated in three unique geometric conformations: square, hexagonal, and near-square (300 nm spaced pits in square pattern, but with AE50 nm disorder). Immunofluorescent labeling of vinculin allowed HOB adhesion complexes to be visualized and quantified by image software. Perhipheral adhesions as well as those within the perinuclear region were observed, and adhesion length and number were seen to vary on nanopit substrates relative to smooth PC. S-phase cells on experimental substrates were identified with bromodeoxyuridine (BrdU) immunofluorescent detection, allowing adhesion quantification to be conducted on a uniform flattened population of cells within the S-phase of the cell cycle. Findings of this study demonstrate the disruptive effects of ordered nanopits on adhesion formation and the role the conformation of nanofeatures plays in modulating these effects. Highly ordered arrays of nanopits resulted in decreased adhesion formation and a reduction in adhesion length, while introducing a degree of controlled disorder present in near-square arrays, was shown to increase focal adhesion formation and size. HOBs were also shown to be affected morphologicaly by the presence and conformation of nanopits. Ordered arrays affected cellular spreading, and induced an elongated cellular phenotype, indicative of increased motility, while near-square nanopit symmetries induced HOB spreading. It is postulated that nanopits affect osteoblast-substrate adhesion by directly or indirectly affecting adhesion complex formation, a phenomenon dependent on nanopit dimension and conformation. ß

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“…Layers thus prepared were then analyzed microscopically. Confocal laser scanning microscopy (CLSM) has significantly advanced the analysis of proteins in intact porous supports (Table 2) and researchers in protein chromatography were forerunners in this respect [62][63][64][65][66][67][68][69][70]. CLSM offers spatial resolution at a length scale well suitable for the characterization of enzyme immobilizates and it is conveniently used even by nonspecialists.…”

Section: Direct Visualization Of Protein Distribution In Solid-suppormentioning

confidence: 99%

Advanced characterization of immobilized enzymes as heterogeneous biocatalysts

2016

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Analysis of protein adsorption characteristics to nano-pore silica particles by using confocal laser scanning microscopy

Cited by 61 publications

References 16 publications

Effect of phase composition and microstructure of calcium phosphate ceramic particles on protein adsorption

Effect of phase composition and microstructure of calcium phosphate ceramic particles on protein adsorption

Regulation of implant surface cell adhesion: characterization and quantification of S‐phase primary osteoblast adhesions on biomimetic nanoscale substrates

Advanced characterization of immobilized enzymes as heterogeneous biocatalysts

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