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PubMed1992

Surface-immobilized polyethylene oxide for bacterial repellence.

Polyethylene terephthalate films were surface-modified with polyethylene oxide (18,500 g/mol) using a solution technique described previously. These films were investigated for their resistance to bacterial adhesion. Three bacterial strains most commonly associated with implant infections, Staphylococcus epidermidis, Staphylococcus aureus and Pseudomonas aeruginosa, were cultured in tryptic soya broth, human plasma and human serum on the polymeric substrates. Significant reductions (between 70 and 95%) in adherent bacteria were observed on the polyethylene oxide-modified substrates compared to the untreated control polyethylene terephthalate. Surface modification with polyethylene oxide may reduce the risk of implant-associated infections. Plasma fibrinogen was observed to play an important role in the adhesion of all three of these species on both the polyethylene oxide-modified and control polyethylene terephthalate materials.

Bacterial Adhesion↗

PubMedDate not supplied

Decreased bacterial adherence to silver-coated stent material: an in vitro study.

Bacteria are important in causing biliary stent blockage through adherence and subsequent biofilm formation. In our in vitro system, surface modification using test polyurethane discs with silver coating led to a reduction in the number of adherent bacteria compared with untreated controls by 10- to 100-fold in an apparently dose-related manner. The effect was more marked in the presence of bile. These results suggest that silver coating may have a potential benefit in preventing stent blockage.

Bacterial Adhesion↗

PubMed1991

[Capsular sack implantation of heparin-modified posterior chamber lenses].

In a prospective study, heparin surface modified posterior chamber lenses were implanted into the capsular bag in 50 patients. The exact positioning of the biconvex, one-piece lens in the bag is the prerequisite for good centration and for an optimal result. The hydrophilisation of the originally hydrophobic PMMA material is achieved by heparin surface modification and results in less inflammation. The aim of this study was to investigate the postoperative behavior of the HSM posterior chamber lens implanted in the bag, with special consideration given to intraocular inflammation. In the postoperative follow-up time of one to seven months, hundred percent of the patients with physiological fundus reached a postoperative visual acuity of 0.7 or better.

Aged↗

PubMed1984

Effect of surface composition on triolein hydrolysis in phospholipid vesicles and microemulsions by a purified acid lipase.

Sonicated dispersions of egg yolk phosphatidylcholine and triolein as vesicles and microemulsions have been used as substrates for the assay of a purified acid lipase. Previous studies have also shown that triolein localized in the surface phase of emulsions is the preferred substrate. In this study, we examined enzyme activity following several surface modifications using both vesicles and microemulsions. When the acidic phospholipids phosphatidylserine and phosphatidic acid were incorporated into both vesicles and microemulsions at up to 10 mol % of the total phospholipid, a dose-dependent reduction in the apparent Km was observed. Using the vesicles as substrate, a dose-dependent decrease in Vmax was also observed. Agarose gel electrophoresis was used to verify suspected changes in net particle charge. Analogous inclusion of phosphatidylethanolamine, sphingomyelin, or cholesterol did not affect kinetic parameters. Addition of oleic acid to sonication mixtures produced vesicles with a decreased apparent Km and Vmax, but triolein hydrolysis in microemulsions was not significantly altered. Triolein-containing vesicles prepared by using dimyristoyl- or dipalmitoylphosphatidylcholine were hydrolyzed maximally at the gel liquid-crystalline transition temperatures of the appropriate phospholipid. Differential scanning calorimetry was used to verify the temperatures of transition in these vesicles. The results indicate that acid lipase activity is influenced by the charge or physical state of the surface phase of model substrates and suggest that degradation of core components of naturally occurring substrates such as lipoprotein may be influenced by chemical changes on the surface of these particles.

Animals↗

PubMed2026

Targeted Nanoparticle Delivery CRISPR/Cas9: overcoming biological barriers, enhancing stability, and improving therapeutic precision.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) has emerged as a promising gene-editing platform for genetic disorders; however, its in vivo application remains limited by low delivery efficiency and biological barriers. Many CRISPR payloads fail to reach target sites due to extracellular degradation, immune clearance, and intracellular trafficking limitations. This review examines the interplay between biological barriers and nanoparticle engineering strategies for CRISPR/Cas9 delivery. A barrier-oriented engineering approach is proposed as a central framework, encompassing ligand-based surface modification for enhanced targeting and uptake, improved circulation stability via PEGylation and biomimetic coatings, and optimized payload release through endosomal escape strategies. Stimulus-responsive nanoparticle systems further enable spatiotemporal control over payload release. Nuclear targeting strategies, including optimization of nuclear localization signals (NLS) and exploitation of endogenous trafficking pathways, are highlighted as key factors for improving genome-level editing efficiency. Despite these advances, major challenges-including limited intracellular delivery efficiency, insufficient targeting precision, and safety concerns-continue to hinder clinical translation. Future directions highlight artificial intelligence-driven nanoparticle design, personalized delivery systems, and next-generation CRISPR platforms. Overall, an integrated, barrier-oriented engineering strategy is essential for advancing CRISPR/Cas9 delivery toward clinical applications, ultimately advancing global good health and well-being.

CRISPR/Cas9↗

PubMed1992

In vitro evaluation of biocompatibility of surface-modified poly(methyl methacrylate) plate with rabbit lens epithelial cells.

Collagen type I was immobilized onto a poly(methyl methacrylate) (PMMA) plate by covalent bonding following surface modification by two methods. One method introduced amino groups by aminolysis with N-lithioethylenediamine (PMMA-NH2) and the other introduced carboxyl groups by graft copolymerization of acrylic acid (AAc) and acrylamide (AAm) (PMMA-COOH). Lens epithelial rabbit cells were cultured on the PMMA plate which was immobilized with collagen. Polygonal cells with a mosaic appearance were observed on the PMMA-COOH plate immobilized with collagen type I, whereas pleomorphic cells were present on the virgin PMMA and on the PMMA-NH2 plate immobilized with collagen type I. We concluded the PMMA-COOH plate immobilized with collagen type I provided a more comfortable atmosphere for lens epithelial cells, causing no metaplasia, than the other plates used in this cell culture model experiment.

Animals↗

PubMed1981

Preparation and application of a photoreactive thrombin analogue: binding to human platelets.

alpha-Thrombin has previously been shown to bind to specific, saturable glycoproteins on the platelet surface. Modification of the thrombin active site with tosyllysyl chloromethyl ketone (TosLysCH2Cl) does not alter thrombin's binding characteristics. Interaction of alpha-thrombin with high-affinity binding sites (KD = 10(-9) M) initiates the platelet response which involves proteolytic hydrolysis of this glycoprotein. Although TosLysCH2Cl--thrombin binds to and competes for the same sites as alpha-thrombin, it cannot induce platelet stimulation because it is enzymatically inactive. In this study, we describe the preparation and application of photoreactive tritium-labeled thrombin analogues. The alpha-thrombin derivative retains its platelet-stimulating and enzymatic activities and, upon photoactivation, covalently binds to specific platelet membrane components. When freshly washed human platelets are exposed to less than saturation doses (less than or equal to 2 nM) of the thrombin derivatives in the dark and photoactivated, a single labeled complex is detected. The same experiment with greater than saturating doses (greater than or equal to 20 nM) of the thrombin derivative yields a similar complex as well as two additional ones. Molecular weight estimates of these thrombin-bound complexes were obtained by gel filtration and NaDodSO4--polyacrylamide gel electrophoresis. The low dose (high affinity) complex with TosLysCH2Cl--thrombin has an approximate molecular weight of 200 000, while that with active alpha-thrombin is smaller, approximately 120 000, due to enzymatic cleavage. The additional complexes detected with the high thrombin dose had estimated molecular weights of 400 000 and 46 000, respectively, and appeared to be the same for TosLysCH2Cl--thrombin and for the alpha-thrombin coupled platelets. These isolated complexes appear to correspond to the two previously detected populations of thrombin binding sites on the platelet.

Azides↗

PubMed1981

The ultrastructure of the thin limbs of Henle in Kidneys of the desert heteromyid (Perognathus penicillatus).

The thin limbs of both long- and short-looped nephrons in Perognathus kidneys were studied with transmission and scanning electron microscopy. The superficial nephrons have a short thin limb located in the vascular bundles of the outer medulla and are characterized by a simple, low-lying epithelium (0.4 +/- 0.1 mu thickness). In contrast, the first descending part of the thin limb of the majority of midcortical and juxtamedullary nephrons has a relatively thick epithelium (1.7 +/- 0.6 mu in thickness) with marked lateral and basal interdigitation and a dense surface covering of microvilli. The remaining part of the long descending thin limb is relatively simple with a low-lying epithelium (0.6 +/- 0.1 mu in thickness), decorated on its surface by sparse microplicae. The bend of the loop and the ascending limb are covered by a very simple low-lying epithelium (0.6 +/- 0.2 mu in thickness) with relatively little surface modification. The extreme urine-concentrating ability of Perognathus does not appear to be due to the development of a unique thin loop epithelium but rather to the extensive length of the inner and outer medulla.

Animals↗

PubMed1975

Transmission and scanning electron microscope studies of calcified cartilage resorption.

The authors' previous report (Savostin-Asling and Asling, '73) demonstrated that Meckel's carilage is a favorable site for study of calcified cartilage resorption. In the present study the ultrastructural features at this resorption front have been examined by transmission and scanning electron microscopes (19-day rat retus). Multinucleated giant cells chondroclasts) dominated the erosion front. The many features which they showed in common with osteoclasts included abundant mitochondria, vacuolation, lysonsomes, sparsity of rough-sufaced endoplasmic reticulum, and deep infoldings at loci of contact with calcified matrix. Crumbling of matrix (with mineral crystals penetrating between these foldings) and fragmentation of collagen fibrils were also seen. The propensity of chondroclasts for spanning several opened lacunae provided special opportunity to demonstrate cell surface modifications in presence or absence of matrix contact. Amebiod processes extending into lacunae were seen by both transmission and scanning procedures; they were sometimes tipped with a veil of filamentous processes as small as 0.3 mum in diameter. Most hypertrophic chondrocytes. when released from lacunae, appeared to be disintegrating. However, in accord with previous evidence of their possible merger with chondroclasts (in light microscopic studies) there was also evidence for breakdown of cell walls between a chondroclast and a chondrocyte in intimate contact, with possibility of cytoplasmic continuity.

Animals↗

PubMed1976

A scanning electron microscope study of the interstitial tissue of the canine testis.

Scanning electron microscopy (SEM) is a potent tool that is especially valuable in interpreting the three-dimensional relationships of cells within tissues. This type of information is obtainable from thin sections in transmission electron microscopy (TEM) only by reconstructions of serial sections. The arrangement of the interstitial cells of the testis in relation to the capillaries and lymphatic channels, in particular, is easier to visualize in SEM than in TEM. Cytoplasmic constituents, as well as cell surface modifications, are demonstrable by this technique. The presence of droplets, presumably lipid droplets, both within and on the Leydig cells and the lymphatic endothelial cells, is quite evident. Other cytoplasmic structures are also apparent. For example, the possible functional significance of "openings" that are seen by SEM on the septa that surround lipid droplets is discussed relative to the appearance of the same area as seen in thin sections or in freeze-fracture replicas. SEM should become a very useful method for studying cytological and morphological alterations that occur in testicular tissue that is subjected to physical or chemical manipulation.

Animals↗

PubMed1980

A new organic solvent for use in the clearing of tissues. I. Soft tissue histology.

Histosol is a non-flammable solvent mixture of synthetic aromatic hydrocarbons with a flash point of 124 degrees F (T.C.C.). It has a lower vapor pressure and evaporation rate than other organic solvents, such as xylene, routinely used as clearing and deparaffinizing agents. Although both xylene and Histosol clear and deparaffinize soft organ tissues effectively in the preparation of permanently mounted stained slides, Histosol appears, in many instances, to be the choice solvent: tissues are easier to section; cell borders and cell surface modifications are most distinct; cytoplasmic eosinophilia is more vivid; and nuclear detail is improved. Of prime importance, Histosol is a safer and more efficient solvent for use in histological and pathological laboratories.

Animals↗

PubMedDate not supplied

Two-dimensional gel electrophoretic analysis of rat sperm membrane interaction with cauda epididymal fluid.

Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used to analyze the polypeptide composition of rat cauda epididymal fluid, blood serum and membrane-enriched fractions of caput, corpus, and cauda epididymal spermatozoa. Several polypeptides were found in both cauda fluid and blood serum, and in both cauda fluid and epididymal spermatozoa. Prominent cauda epididymal fluid polypeptides that were associated with caput, corpus, and cauda sperm membranes were 32 and 33 kDa. Passage of spermatozoa from the caput to the cauda epididymidis was characterized by the loss of three glycopolypeptides of 32, 30 and 29 kDa, and by the addition of a 37-kDa glycopolypeptide. Incubation of intact caput, corpus and cauda spermatozoa with cauda epididymal fluid revealed major changes in the polypeptide maps of the incubation fluid and the membrane-enriched fractions of caput and corpus, but not cauda spermatozoa. The incubation of cauda fluid with caput and corpus sperm cells was characterized by a loss of several polypeptides and the addition of a 24-kDa glycopolypeptide. The most striking change in spermatozoa incubated with cauda epididymal fluid was the addition of two glycopolypeptides of 32 and 33 kDa to the polypeptide maps of caput sperm cells. These data demonstrate that rat spermatozoa undergo surface modifications during epididymal maturation and that these modifications can be influenced by epididymal fluid.

Animals↗

PubMed1991

Asymmetric membrane filters for the removal of leukocytes from blood.

As part of a study on the mechanisms of leukocyte filtration, the influence of pore size distribution on filter efficiency was investigated. Conventional leukocyte filters are not suitable for model studies, as these filters are composed of tightly packed synthetic fibers, with a poorly defined porous structure. Therefore, open cellular polyurethane membranes with pore size distributions varying from approximately 15 to 65 microns were prepared. Filtration experiments with stacked packages of these membranes showed that leukocytes are best removed (greater than 99%) by filters with a pore size distribution of 11-19 microns. These pore sizes approach the size of leukocytes (6-12 microns). However, due to fast clogging, blood flow through these filters is rapidly reduced, which results in a low filter capacity. With an asymmetric membrane filter, in which the pore size decreases from about 65 to 15 microns in the direction of blood flow, both moderate removal of leukocytes (greater than 80%) and maintenance of flow (approximately 0.2 mL/s) are obtained. This results in efficient leukocyte removal. From cell analysis of both filtrate and filter, it is concluded that adhesion rather than sieving is the major filtration mechanism. Thus, further optimization of the filter may be achieved by surface modification.

Cell Separation↗

PubMed1992

Albumin-binding surfaces for implantable devices.

Surfaces of implantable and blood contact-devices accumulate adsorbed and denatured proteins. This anomalous layer of proteins may help trigger unwanted events such as activation of coagulation systems and, perhaps, chronic inflammation. Because, in many experimental systems, the purposeful coating of surfaces with albumin will biologically "passivate" materials, we have attempted to develop polymers which, when exposed to blood or body fluids, will spontaneously, selectively, and reversibly adsorb host albumin. We report here a novel derivatization technique for increasing the albumin affinity of implantable polyetherurethane (PU). The technique is based on the incorporation of high-molecular-weight dextran to which the albumin-binding dye Cibacron Blue is covalently attached. Somewhat surprisingly, the amounts of human albumin adsorbed by Blue Dextran-modified and unmodified PU are quite similar. There are, however, important differences. First, the binding of albumin to derivatized PU is specific and not readily blocked by proteins in albumin-depleted human serum. Second, the majority of albumin associated with derivatized PU appears to be reversibly bound. Third, the binding of albumin to derivatized PU evidently is mediated primarily through ligand-specific binding of the protein to the albumin-binding dextran-dye conjugate. We conclude that it is possible to produce implantable polymers having surfaces which display albumin-binding dyes that selectively and reversibly bind albumin. Materials with this property, when implanted or exposed to blood, should form an infinitely renewable coating of albumin derived from physiologic fluids. This surface modification strategy may spawn a new generation of implantable materials with improved biologic compatibility.

Adsorption↗

PubMed1976

Cytological observations of the ovarian epithelium in mammals during the reproductive cycle.

We have studied the ovarian epithelijm at various stages of the reproductive cycle in a number of mammalian species utilizing light microscopy, scanning microscopy, the freeze-fracture technique, transmission microscopy and by employing specialized tracers that use lanthanum and horseradish peroxidase. We found that the epithelial cells are joined by incomplete tight junctions, gap junctions, and desmosomes. The cytoplasmic matrix contains a large irregularly shaped nucleus, few microtubules, microfilaments, mitochondria, endoplasmic reticulum and a host of coated and non-coated vesicles of varying diameters. The saccules comprising the large Golgi complex and its companion vesicles are associated with a basal body-centriole complex: some of these saccules and affiliated vesicles are acid phosphatase positive. Surface modifications of ovarian epithelial cells include numerous microvilli, some of which have a bulbous tip, and plications of the lateral plasma membrane which are thought to accomodate volume changes of the ovary during follicular development. Many coated and non-coated endocytotic caveolae were found on these cells, particularly in the basal area. These caveolae internalized exogeneously administered horseradish peroxidase. We view the marked endocytotic activity as an efficient transport mechanism for partially removing substances from the interstitium of the ovary and the peritoneum.

Animals↗

PubMed1985

Fine structure of the epithelia of the vomeronasal organ of horse and cattle. A comparative study.

The vomeronasal organ of both horses and cattle is a tubular structure situated bilaterally at the base of the nasal septum. In frontal plane the shape of its lumen is semilunar to crescent. The sensory epithelium lining the medial wall of the lumen contains receptor, supporting and basal cells with some surface modifications in both species. In the horse, a structure similar to a microprocess was observed among the microvilli of receptor cells. In cattle, a large mass of the cytoplasm of the receptor cell occasionally protrudes to form a bleb-like structure. The supranuclear cytoplasm of the receptor cells contain mitochondria, free ribosomes, rough endoplasmic reticula, Golgi apparatus, lysosomes and multivesicular bodies. Some receptor cells were pyknotic. In both species the respiratory epithelia of the lateral wall of the lumen contain ciliated, non-ciliated and basal cells. In the horse, this epithelium differs from that of other species in evidence of prominent secretory function.

Animals↗

PubMed1982

Studies on the development of extra-endothelial and intra-endothelial pigment deposits by means of direct and indirect contact specular microscopy of the cornea.

Retro-corneal pigmentation as a result of primary (degenerative) or secondary (inflammatory, traumatic) pigment dispersion represents one of the most frequent findings in specular microscopy. The analysis of the pigment distribution patterns of dense deposits like those in Krukenberg spindle (KS) permits the postulation of a hypothesis concerning the development of such typical corneal pigmentations and the pigmentation of the posterior surface of the cornea in general, such as diffuse pigment dispersion (Vogt 1930), which is seen frequently. In a particular case of double KS with extreme myopia and cataract, endothelial morphological findings were documented over a period of 1 year, both before and after cataract extraction. This was done by means of contact and non-contact specular microscopy in various spindle areas. The morpho-metric studies (cell density/mm2; cell perimeter; largest smallest and average cell diameter; specific surface; horizontal and vertical cell orientation) were carried out automatically using a computerised video image analyser. The changes in the area of densest pigmentation in the KS centre confirm the histologically proven capacity of pigment phagocytosis by human corneal endothelium. These findings indicate that in specular microscopy, both endothelial cellular surface modifications and intra-cellular changes are documentable. The extracellular pigment deposits can also be shown by means of 'indirect' specular microscopy, the optical principle of which is described.

Aged↗

PubMed1990

Enzyme activities of lung lavage in silicosis.

The cytotoxic effect of quartz on lung cells has been well documented by in vitro and animal studies, but the pertinence of these findings to humans has not yet been documented. We measured lactate dehydrogenase (LDH) activities in the lung lavage of 24 long-term workers in the Québec granite industry and 25 control subjects. We found significant increases in LDH activities in the workers' lung lavage, even in the absence of established silicosis (9 subjects). We looked at a similar observation in the sheep model of early silicosis, measured quartz content of lung lavage, and found significant correlation with LDH levels (R = 0.64, p less than 0.001). All of the quartz particles in human and sheep lung lavage were in the alveolar macrophages. To test further the relationship of macrophage damage (cytotoxicity of quartz) we measured the release of LDH by sheep alveolar macrophage in 24 h cell culture under control conditions, exposure to inert dust, titanium, minusil-5 quartz, or aluminum-treated quartz. The LDH release was at control levels during titanium exposure and showed a significantly dose-related increase during quartz exposure. The latter cytotoxic effect was largely attenuated by aluminum treatment of quartz. These in vitro data agreed with previous reports. This study presents evidence of a cytotoxic effect of quartz inhalation in humans. The effect is related to the intensity of quartz retention in the lung macrophages; it is not a nonspecific dust exposure effect and can be attenuated by surface modification of the quartz.

Adult↗