Colonization of n-butyl-2-cyanoacrylate tissue adhesive by Staphylococcus epidermidis

In vitro adhesion of Staphylococcus epidermidis to cyanoacrylate tissue adhesive surfaces was investigated by employing a modified Robbins device. Bacterial colonization of n-butyl-2-cyanoacrylate tissue adhesive polymer surfaces was demonstrated by employing scanning electron microscopy, epifluorescence technique, and aerobic plate counts. The S. epidermidis was found to rapidly colonize the n-butyl-2-cyanoacrylate polymer surface producing a biofilm composed of embedded bacteria in an extensive amorphous matrix that totally occludes the material surface. The results indicate that the bacteriocidal properties of the polymer to this bacteria are weak or absent.     

合成α-氰基丙烯酸正丁酯医用胶用于肝脏止血

BACKGROUND: There are chemosynthetic and biological medical adhesives, and the former mainly composed by cyanoacrylate, holds poor hydrophilicity and is easy to break off, thereby achieving unsatisfactory hemostatic effect. OBJECTIVE: To evaluate the hemostatic and adhesion effect of the modified α-N butyl cyanoacrylate (NBCA) by the hemostatic test in white rabbit livers. METHODS: Material imprvement: cyanoacrylate monomer was prepared and its components and contents were analyzed by infrared spectrum and gas chromatograph. Sodium lauroyl sarcosinate was used as surfactant to improve the cohesive force between NBCA and tissues and increase the hydrophility. Hemostatic test in the liver incision: the liver incision of rabbits in three groups were sewn with absorbable suture and then covered with gelatin sponge (control group), cyanoacrylate and NBCA medical adhesives, respectively. Blood loss and bleeding time were recorded within 20 minutes. Hemostatic test in the liver section: the liver sections in two groups were smeared with surfactant plus NBCA and cyanoacrylate, respectively. The gross appearance of wound and its adherence to the omentum majus, as well as the histological changes by hematoxylin-eosin staining were observed at 1, 10 and 20 days postoperatively. RSULTS AND CONCLUSIPON: The adhesiveness of NBCA medical adhesive was excellent. Infrared spectrum and gas chromatograph found that the main component was NBCA up to 84.56%, with 8.2% reactants and 3.3% byproducts. The modified NBCA medical adhesive significantly decreased the blood loss and bleeding time compared with the control group (P < 0.01), and did not significantly differ from the cyanoacrylate (P > 0.05). In addition, the modified NBCA adhered more firmly to the wound, and was absorbed completely at 20 days postoperatively, showing no adverse reaction. These results suggest that NBCA is better and more stable than gelatin sponge, and exerts overt hemostatic effect causing no adverse reaction.     

Durability of the bond between bone and various 2-cyanoacrylates in an aqueous environment.

The durability of the bond strength developed between 2-cyanoacrylate esters and bone has been determined by aging specimens in water. One-day bond strength of the isobutyl and isomeric amyl 2-cyanoacrylates varied from 6.2 to 7.2 MPa. The strength of the bond decreased on storage or on thermocycling in water. Hydrolytic stability increased with increasing length of the alkyl ester group. After a six-month storage in water the various amyl 2-cyanoacrylates retained from 70% to 73% of their one-day bond strength. Pretreatment of the bone surface prior to application of the adhesive did not prove beneficial. The cured 2-cyanoacrylate can be removed from the substrate surface by appropriate solvents. Thus, it is not bonded covalently to bone. The bond strength, especially of the isobutyl and amyl 2-cyanoacrylates to bone in an aqueous environment, appears to be superior to other adhesives. Provided these monomers are biocompatible, they may be useful clinically where an intermediate-term adhesion is desired.     

Adhesiveness of dental resin-based restorative materials investigated with atomic force microscopy

This study aimed to show that the polymerization contraction of dental methacrylate-based materials, when used as adhesives on hard substrate, produces voids at the material-substrate interface. This phenomenology is closely related with the nanoleakage and the sealing ability of these materials. One prime/bond system, three restorative composite resins, and one orthodontic bonding system were cured by using mirror-like glass slides as a compliance-free reference substrate. The adhesive surface was analyzed by atomic force microscopy, and the polymerization contraction of bulk material was tested by laser beam-scanning method. Nanoperiodic structure of three-dimensional (3D) images, section analysis, and roughness characteristics (R(a) and R(z)) indicated that polymerization contraction produced voids at the interface. When the adhesive surface was exposed to oral simulating fluids (water, ethanol, and lactic acid solutions), hydrolytic degradation involved some hundreds of nanometers in depth. In visible light-cured (VLC) materials, the interface porosity decreased when an irradiation pause ( approximately 2 min) was carried out during gelation.     

皮肤粘合剂“爱必肤“用于小儿先天性心脏病手术切口闭合的临床早期观察

目的观察皮肤粘合剂“爱必肤“(EPIGLU,主要成分为2-乙基氰丙烯酸酯)应用于小儿先天性心脏病术后切口的效果.方法将“爱必肤“点涂于先天性心脏病后对合良好的皮肤切口表面共30例,并观察12天.结果29例切口愈合较好或仅有轻微开裂,1例有部分开裂(重新缝合后愈合).4例有伤口渗血,2例较多.所有例数均未见过敏反应,未见皮肤感染.结论皮肤粘合剂“爱必肤“粘合对合良好的皮肤切口效果确实可靠,不需拆线,组织反应小,有广泛的临床应用前景.     

Modulatory role of focal adhesion kinase in regulating human pulmonary arterial endothelial barrier function

The adhesive force generated by the interaction of integrin receptors with extracellular matrix (ECM) at the focal adhesion complex may regulate endothelial cell shape, and thereby the endothelial barrier function. We studied the role of focal adhesion kinase (FAK) activated by integrin signalling in regulating cell shape using cultured human pulmonary artery endothelial cells. We used FAK antisense oligonucleotide (targeted to the 3'-untranslated region of FAK mRNA (5'-CTCTGGTTGATGGGATTG-3') to determine the role of FAK in the mechanism of thrombin-induced increase in endothelial permeability. Reduction in FAK expression by the antisense augmented the thrombin-induced decrease in transendothelial electrical resistance (decrease in mock transfected cells of −43 ± 1 % and in sense-transfected cells of −40 ± 4 %, compared to the decrease in antisense-transfected cells of −60 ± 3 %). Reduction in FAK expression also prolonged the drop in electrical resistance and prevented the recovery seen in control endothelial cells. Thus, the thrombin-induced increase in permeability is both greater and attenuated in the absence of FAK expression. Inhibition of actin polymerization with latrunculin-A prevented the translocation of FAK to focal adhesion sites and tyrosine phosphorylation of FAK and paxillin, and concomitantly reduced the thrombin-induced decrease in electrical resistance by ∼50 %. Thus, the modulatory role of FAK on endothelial barrier function is dependent on actin polymerization. FAK translocation to focal adhesion complex in endothelial cells guided by actin cables and the consequent activation of FAK-associated proteins serve to reverse the decrease in endothelial barrier function caused by inflammatory mediators such as thrombin.     

A study of compatibility between cells and biopolymeric surfaces through quantitative measurements of adhesive forces

The mechanism of cell adhesion to biomaterials or components of the extracellular matrix is an important topic in the field of tissue engineering and related biotechnological processes. Many factors affect cell adhesion, and many biochemical and biological studies have attempted to identify their roles in the adhesion mechanism. Systematic studies of this nature require quantification of the adhesive force of a cell to identify the effect of a specific factor. However, most studies of cell adhesive force have used qualitative approaches. We propose a new technique for quantifying the force by which cells adhere to various biomaterial surfaces, which utilizes the relationship between the deflection of a cantilever beam and the required force. A micropipette was used as the cantilever beam. This technique was used to measure the attachment forces of chondrocytes seeded on three different biodegradable polymers commonly used in tissue engineering and medicine: poly epsilon-carprolactone (PCL), poly(L-lactide) (PLLA) and poly(lactic-co-glycolic acid) (PGLA, L/G = 75:25). The bond between the cells and the three polymers was evaluated using the quantified adhesive forces. The adhesive forces were also measured 8, 12, 24 h and 5 days after seeding the chondrocytes on the polymer surfaces. Results of statistical analysis showed that the cells attached to the PLLA had the strongest average attachment force for up to 24 h after seeding (P < 0.05).     

Preparation of thermoresponsive polymer brush surfaces and their interaction with cells

Poly(N-isopropylacrylamide) (PIPAAm) brush surfaces with different layer thickness on polystyrene substrates were prepared by surface-initiated atom transfer radical polymerization (ATRP). Surface characteristics of PIPAAm brushes and their influence on adhesion and detachment of bovine carotid artery endothelial cells (ECs) were controlled by PIPAAm layer thickness. Surface hydrophilicity increased with PIPAAm layer thickness at 37 degrees C because PIPAAm brush surfaces with higher thickness provide more extended chain conformations with relatively high chain mobility, and accompanying polymer chain hydration. These surface property alterations lead to negligible cell adhesion through minimal matrix protein adsorption and also modified surface modulus. By adjusting polymerization reaction conditions and time, polymer layers supporting confluent cultures of ECs were possible. Confluent EC monolayers spontaneously detached as contiguous cell sheets from PIPAAm brush surfaces at reduced temperatures. Thermoresponsive cell adhesion and detachment behavior were analyzed from the standpoint of surface physicochemical characteristics. Thermoresponsive surfaces prepared by surface-initiated ATRP techniques allow surface selection in preparing cell sheets from attachment-dependent cells having relatively strong adhesive property for tissue engineering applications.     

ProNectin F-grafted-ethylene vinyl alcohol copolymer (EVAL) as a liquid type material for treating cerebral aneurysm--an in vivo and in vitro study

The authors aimed to develop a liquid material for embolization of aneurysms. In vitro and in vivo performances of the new embolic material were examined by cell culture and using an aneurysm model made in common carotid arteries (CCAs) of adult rats. Engineered protein ProNectin F (PnF), which contains 13 sites of an arginine-glycine-aspartic acid (RGD), was grafted onto ethylene vinyl alcohol copolymer (EVAL-g-PnF). The liquid material, EVAL-g-PnF dissolved in DMSO, was infused into an aneurysm model. The blood segments were harvested 2, 5, and 14 days and examined histologically. A number of bovine coronary artery endothelial cells became able to attach to and form cobblestone-like islands on the EVAL by incorporating PnF. The aneurysm model infused with the EVAL-g-PnF solution revealed that the aneurysm lumen was filled with proliferated fibroblasts and macrophages. On the other hand, the aneurysm model treated with unmodified EVAL showed that the cavity was almost filled with EVAL mass and that fibroblasts and macrophages filled a narrow space between the EVAL mass and the cavity wall. The results indicate that EVAL-g-PnF could be more suitable for reorganizing the cavity of an aneurysm than native EVAL.     

The effect of different power densities and method of exposure on the marginal adaptation of four light-cured dental restorative materials

PURPOSE: To determine whether marginal adhesion is sensitive to different irradiation parameters, we investigated the in vitro adhesion values of four dental resins on metal surfaces. METHODS: Four groups of eight specimens each of Z250, Filtek flow, Dyract AP and Dyract flow were placed in pre-treated stainless steel cavities and irradiated using different methods of exposure. The curing lights used were a Spectrum 800 halogen curing light at settings of 800 and 450 mW/cm(2) and an Optilux 501 ramping light. The maximum amount of push-out force required to displace the resin from the metal cavity was equated with adhesive value (shear bond strength). Comparisons (ANOVA, p<0.0001) were made within the same material and between the different materials when using different curing protocols. RESULTS: Significant lower bond strengths were recorded when curing was done by gradually increasing the intensity (ramping method) compared to curing with the fixed intensities (p>0.0001) Comparing the fixed intensities, significant lower bond strength values were obtained at 800 mW/cm(2) compared to 450 mW/cm(2) (p<0.0001). For all exposures, the two flowable materials demonstrated weaker values when compared to the higher filled materials. CLINICAL SIGNIFICANCE: The advantage of initial slow polymerization (more elasticity and less tension) obtained by the so-called "soft start" method, was offset by a rise in total polymerization shrinkage, when final curing was completed at 1130 mW/cm(2). These tests demonstrated that using halogen units, exposure for 40s with an intensity of 450 mW/cm(2) appeared to be the most promising for light-curing dental resin composites.     

A study of compatibility between cells and biopolymeric surfaces through quantitative measurements of adhesive forces

The mechanism of cell adhesion to biomaterials or components of the extracellular matrix is an important topic in the field of tissue engineering and related biotechnological processes. Many factors affect cell adhesion, and many biochemical and biological studies have attempted to identify their roles in the adhesion mechanism. Systematic studies of this nature require quantification of the adhesive force of a cell to identify the effect of a specific factor. However, most studies of cell adhesive force have used qualitative approaches. We propose a new technique for quantifying the force by which cells adhere to various biomaterial surfaces, which utilizes the relationship between the deflection of a cantilever beam and the required force. A micropipette was used as the cantilever beam. This technique was used to measure the attachment forces of chondrocytes seeded on three different biodegradable polymers commonly used in tissue engineering and medicine: poly ε-carprolactone (PCL), poly(L-lactide) (PLLA) and poly(lactic-co-glycolic acid) (PGLA, L/G = 75 : 25). The bond between the cells and the three polymers was evaluated using the quantified adhesive forces. The adhesive forces were also measured 8, 12, 24 h and 5 days after seeding the chondrocytes on the polymer surfaces. Results of statistical analysis showed that the cells attached to the PLLA had the strongest average attachment force for up to 24 h after seeding (P < 0.05).     

Axisymmetric adhesion tests of soft materials

We describe a general, linearized fracture mechanics analysis for studying the adhesive properties of elastic, low modulus materials. Several adhesion tests are described, but all involve an elastic material which is brought into contact with a rigid surface along an axis of radial symmetry. Relationships between the load, displacement, and radius of the circular contact area between the two materials are described. These relationships involve the elastic modulus of the compliant material, the energy release rate (or adhesion energy) and various parameters which characterize the geometry of interest. The ratio of the contact radius to the thickness of the elastic material is shown to be a particularly important parameter. After reviewing some general concepts relevant to the adhesion of soft polymeric materials, we describe the fracture mechanics analysis, and provide examples from our own work on the adhesion of elastomers, thermoreversible gels and pressure sensitive adhesives.     

Sub-micron texturing for reducing platelet adhesion to polyurethane biomaterials

Platelet adhesion is a key event in thrombus development on blood-contacting medical devices. It has been demonstrated that changes to the chemistry of a material surface can reduce platelet adhesion. In this work, it is hypothesized that sub-micron surface textures may also reduce adhesion via a decrease in the surface area of material with which platelets can make contact, and hence a decreased probability of interaction with adhesive ligands. A polyether(urethane urea) was textured with two different sizes of sub-micron pillars using a replication molding technique that did not alter the material surface chemistry. Adhesion of platelets was assessed in a physiologically relevant shear stress range of 0-67 dyn/cm2 using a rotating disk system. Platelets were immunofluorescently labeled and adhesion was compared on smooth and textured samples. Platelet adhesion was greatest at low shear stress ranging from 0 to 5 dyn/cm2, and sub-micron textures were observed to reduce platelet adhesion in this range. Additionally, non-adherent platelets did not demonstrate large-scale activation after exposure to textured samples. We conclude that surface textures with sub-platelet dimensions may reduce platelet adhesion from plasma to polyether(urethane urea) at low shear stress.     

Effects of intermittent hydrostatic pressure on cell adhesive forces and other related parameters under various resting periods

The purpose of this study is to investigate the effects of intermittent hydrostatic pressure with various resting periods on the cell adhesive forces and other parameters related to spreading in early stage of cell adhesion. For this, bovine pulmonary arterial endothelium (CPAE, cell line), porcine articular chondrocytes, and human endothelial cells (HECs) were used. The cells were divided into six different experimental groups. Control group was cultured without stimulation, while the constant pressure was applied to group 1 for 2 h. Groups 2-5 were intermittently pressurized for 2 min at a time over a 2-h period with 5, 10, 15, and 20-min resting periods, respectively. Each group was then split into two subgroups, depending whether it experienced extra 60 min stabilization period after stimulation. The average adhesive force and the number and area of focal contacts were significantly higher in the group 4 subgroup, which received an extra 60 min of culture than in the other groups. Similarly, other parameters in this subgroup were significantly different from those in the other groups. The focal contact area and adhesive force were closely related (r = 0.990). We concluded that the mechanical stimuli affect cell adhesion and that the length of the resting period influences the adhesive forces generated at the early stages of adhesion.     

Possible involvement of altered RGD sequence in reduced adhesive and spreading activities of advanced glycation end product-modified fibronectin to vascular smooth muscle cells

Although fibronectin (FN) modified by advanced glycation end products (AGEs) has been shown to contribute to the development of diabetic vascular complications through its reduced adhesive activity to vascular cells, little is known about changes in the cell binding domain of AGE-modified FN. Here we examined the mechanism of reduced adhesive and spreading activities of AGE-modified FN to vascular smooth muscle cells (SMCs), particularly the contribution of modification of Arg-Gly-Asp (RGD) sequence. Incubation with glucose caused not only the formation of N(epsilon) -carboxymethyllysine and pentosidine, but also polymerization of FN in a dose- and time-dependent manner. AGE-modified FN had significantly low adhesive and spreading activities to cultured SMCs. On the other hand, multimeric FN formed by disulfide bonds did not show any effect on either cell adhesion or spreading. The adhesive activity of type I collagen, one of the RGD sequence-containing proteins, to SMCs also decreased by AGE-modification. The inhibitory effect of AGE-modification on cell adhesion was significantly greater in type I collagen than in FN. Although the extent of AGE-modification of type I collagen was indistinguishable from that of FN, AGE-modification decreased the arginine content of type I collagen by 69.5% and of FN by 30.6%, compared with their non-glycated forms. The addition of RGD peptides caused a decrease in adhesion of SMCs to non-glycated FN, but not to AGE-modified FN. Modification of RGD sequence with glyoxal eliminated its inhibitory effect on cell adhesion. Our results suggest that a marked decrease in adhesive and spreading activities of AGE-modified FN to SMCs might largely be due to a modification of its RGD sequence by AGE, thus suggesting a potential link between AGE modification of FN and the pathogenesis of diabetic angiopathy.     

Possible Involvement of Altered RGD Sequence in Reduced Adhesive and Spreading Activities of Advanced Glycation End Product-Modified Fibronectin to Vascular Smooth Muscle Cells

Although fibronectin (FN) modified by advanced glycation end products (AGEs) has been shown to contribute to the development of diabetic vascular complications through its reduced adhesive activity to vascular cells, little is known about changes in the cell binding domain of AGE-modified FN. Here we examined the mechanism of reduced adhesive and spreading activities of AGE-modified FN to vascular smooth muscle cells (SMCs), particularly the contribution of modification of Arg-Gly-Asp (RGD) sequence. Incubation with glucose caused not only the formation of N-carboxymethyllysine and pentosidine, but also polymerization of FN in a dose- and time-dependent manner. AGE-modified FN had significantly low adhesive and spreading activities to cultured SMCs. On the other hand, multimeric FN formed by disulfide bonds did not show any effect on either cell adhesion or spreading. The adhesive activity of type I collagen, one of the RGD sequence-containing proteins, to SMCs also decreased by AGE-modification. The inhibitory effect of AGE-modification on cell adhesion was significantly greater in type I collagen than in FN. Although the extent of AGE-modification of type I collagen was indistinguishable from that of FN, AGE-modification decreased the arginine content of type I collagen by 69.5% and of FN by 30.6%, compared with their non-glycated forms. The addition of RGD peptides caused a decrease in adhesion of SMCs to non-glycated FN, but not to AGE-modified FN. Modification of RGD sequence with glyoxal eliminated its inhibitory effect on cell adhesion. Our results suggest that a marked decrease in adhesive and spreading activities of AGE-modified FN to SMCs might largely be due to a modification of its RGD sequence by AGE, thus suggesting a potential link between AGE modification of FN and the pathogenesis of diabetic angiopathy.     

金黄色葡萄球菌对中性粒细胞粘附特性的影响

目的：研究中性粒细胞在外源病原微生物金黄色葡萄球菌及其分泌物 免疫激活状态下的粘附特性,考察不同的细菌浓度对中性粒细胞粘附性的影响。材料与方法：采用微管吸吮技术测量大鼠中性粒细胞经不同浓度金黄色葡萄菌及其分泌物刺激后与内皮细胞的粘附特性。结果：中性粒细胞经金黄色葡萄球菌免疫激活后与内皮细胞的粘附力和对照组比较均显著增高,但随着细菌浓度的提高而表现出饱和效应。用金黄色葡萄球菌分泌物（培养液）处理中性粒细胞中,其与内皮细胞的粘附力和对照组比较却有所降低。将经细菌处理后的中性粒细胞分离出再与内皮细胞黏附,黏附力增加幅度更大。结论：中性粒细胞经金黄色葡萄球菌免疫激活后粘附分子表达增加,但金黄色葡萄球菌分泌物可能对中性粒细胞粘附分子表达起抑制作用。在此过程中,内皮细胞可能受到金黄色葡萄球菌及其分泌物的损伤。     

An Algorithm for Tracking Microcatheters in Fluoroscopy

Currently, a large number of endovascular interventions are performed for treatment of intracranial aneurysms. For these treatments, correct positioning of microcatheter tips, microguide wire tips, or coils is essential. Techniques to detect such devices may facilitate endovascular interventions. In this paper, we describe an algorithm for tracking of microcatheter tips during fluoroscopically guided neuroendovascular interventions. A sequence of fluoroscopic images (1,024 × 1,024 × 12 bits) was acquired using a C-arm angiography system as a microcatheter was passed through a carotid phantom which was on top of a head phantom. The carotid phantom was a silicone cylinder containing a simulated vessel with the shape and curvatures of the internal carotid artery. The head phantom consisted of a human skull and tissue-equivalent material. To detect the microcatheter in a given fluoroscopic frame, a background image consisting of an average of the four previous frames is subtracted from the current frame, the resulting image is filtered using a matched filter, and the position of maximum intensity in the filtered image is taken as the catheter tip position in the current frame. The distance between the tracked position and the correct position (error distance) was measured in each of the fluoroscopic images. The mean and standard deviation of the error distance values were 0.277 mm (1.59 pixels) and 0.26 mm (1.5 pixels), respectively. The error distance was less than 3 pixels in the 93.0% frames. Although the algorithm intermittently failed to correctly detect the catheter, the algorithm recovered the catheter in subsequent frames.     

Cell adhesion to borate glasses by colloidal probe microscopy

The adhesion of osteoblast-like cells to silicate and borate glasses was measured in cell growth medium using colloidal probe microscopy. The probes consisted of silicate and borate glass spheres, 25-50 μm in diameter, attached to atomic force microscope cantilevers. Variables of the study included glass composition and time of contact of the cell to the glasses. Increasing the time of contact from 15 to 900 s increased the force of adhesion. The data could be plotted linearly on a log-log plot of adhesive force versus time. Of the seven glasses tested, five had slopes close to 0.5, suggesting a square root dependence of the adhesive force on the contact time. Such behavior can be interpreted as a diffusion limited process occurring during the early stages of cell attachment. We suggest that the rate limiting step in the adhesion process is the diffusion of integrins resident in the cell membrane to the area of cell attachment. Data presented in this paper support the hypothesis of Hench et al. that strong adhesion depends on the formation of a calcium phosphate reaction layer on the surfaces of the glass. Glasses that did not form a calcium phosphate layer exhibited a weaker adhesive force relative to those glasses that did form a calcium phosphate layer.     

Val-ala-pro-gly, an elastin-derived non-integrin ligand: smooth muscle cell adhesion and specificity

The elastin-derived peptide val-ala-pro-gly (VAPG) may be useful as a biospecific cell adhesion ligand for smooth muscle cells. By grafting the peptide sequence into a hydrogel material, we were able to assess its effects on smooth muscle cell adhesion and spreading. These materials are photopolymerizable hydrogels based on acrylate-terminated derivatives of polyethylene glycol (PEG). Because of their high PEG content, these materials are highly resistant to protein adsorption and cell adhesion. However, PEG diacrylate derivatives can be mixed with adhesive peptide-modified PEG monoacrylate derivatives to facilitate cell adhesion. Following photopolymerization, PEG monoacrylate derivatives are grafted into the hydrogel network formed by the PEG diacrylate. This results in covalent immobilization of adhesive peptides to the hydrogel via a flexible linker chain. The resistance of PEG to protein adsorption makes it an ideal material for this model system since cell-material interactions are limited to biomolecules that are covalently incorporated into the material. In this case we were able to demonstrate that VAPG is specific for adhesion of smooth muscle cells. It also was shown that fibroblasts, endothelial cells, and platelets cannot adhere to VAPG. In addition, not only was smooth muscle cell adhesion dependent on ligand concentration, but also cell spreading increased with increasing ligand concentration.