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A new approach to end-functionalization of poly(3-hydroxybutyrate)[PHB] is described. Using genetically engineered PHB synthase fused with a 10x-histidine units at its N-terminus, end-functionalized PHB was synthesized and used for the solid surface modification.
Because of the limited mechanical properties of tissue substitutes formed by culturing cells on polymeric scaffolds, other approaches to tissue engineering must be explored for applications that require complete and immediate ability to bear weight, e.g. total joint replacements. Biochemical surface modification offers a way to partially regulate events at the bone-implant interface to obtain preferred tissue responses. Tresyl chloride, gamma-aminopropyltriethoxysilane (APS) and p-nitrophenyl chloroformate (p-NPC) immobilization schemes were used to couple a model enzyme, trypsin, on bulk samples of Co-Cr-Mo. For comparison, samples were simply adsorbed with protein. The three derivatization schemes resulted in different patterns and levels of activity. Tresyl chloride was not effective in immobilizing active enzyme on Co-Cr-Mo. Aqueous silanization with 12.5% APS resulted in optimal immobilized activity. Activity on samples derivatized with 0.65 mg p-NPC cm-2 was four to five times greater than that on samples simple adsorbed with enzyme or optimally derivatized with APS and was about eight times that on tresylated samples. This work demonstrates that, although different methods have different effectiveness, chemical derivatization can be used to alter the amount and/or stability of biomolecules immobilized on the surface of Co-Cr-Mo.
BACKGROUND: Modern implant systems used in the cervical region do not allow for the ingrowth of peri-implant mucosa. The aim of the present study was to use a laser to modify titanium implants used in that region in order to promote mucosa ingrowth, thereby creating a biological barrier against bacterial infection. METHODS: A KrF-excimer laser (lambda=248 nm) was used (tau=30 ns, fmax=50 Hz, Emax=1.2 J). Craters were generated, under normal atmospheric conditions or in vacuum, in the polished regions of 18 Frialit 2 implants. Surface analysis was performed using a scanning electron microscope, an energy dispersive X-ray technique, and profile analysis. RESULTS: By adjusting the parameters used, it was possible to create canals with widths of up to 100 microm and depths of 450 microm. Canal morphology was influenced by the laser energy and the surrounding atmospheric conditions. Irradiation with 3,000 pulses (650 mJ per pulse) resulted in rough surfaces under normal atmospheric conditions, whereas these parameters resulted in smooth surfaces in vacuum. The surface chemistry was also influenced by the laser parameters used. CONCLUSION: Laser-assisted modification of titanium surfaces permits fabrication of 3-D surface modifications. The surface chemistry can also be influenced, depending upon the parameters chosen. Further studies are necessary to evaluate whether ingrowth of mucosa into the canals can be realized.
By using neutron and synchrotron x-ray small-angle scattering techniques, we investigated the process of the complexation of gangliosides with proteins. We treated monosialoganglioside (G(M1)), disialoganglioside (G(D1a)), and a mixture of G(M1)/G(D1a). Proteins used were bovine serum albumins whose surfaces were modified with different sugars (deoxy-D-galactose, deoxy-L-fucose, deoxymaltitol, and deoxycellobiitol), which were used as model glycoproteins in a membrane. We found that the complexation of gangliosides with albumins greatly depends on the combination of ganglioside species and protein surface modification. With a varying protein/ganglioside ratio in a buffer solution at pH 7, the complexation of G(M1) or G(D1a) with albumins modified by monosaccharides appears to be less destructive for ganglioside aggregate structures in forming large complexes; the complexation of G(D1a) with the albumins modified by disaccharides induces the formation of complexes with a dimeric structure; and the complexation of G(M1) with albumins modified by disaccharides, to form small complexes, is very destructive. The present results show a strong dependence of the interaction between ganglioside and protein on the characteristics of the ganglioside and protein surface, which would relate to a physiological function of gangliosides, such as a function regulating the receptor activity of glycoproteins in a cell membrane.
The sample of fossil human bones from the Sima de los Huesos, Atapuerca, has been analysed to trace parts of its taphonomic history. The work reported here is restricted to analysis of the skeletal elements preserved and their surface modifications. Preliminary plans of specimen distribution published 6 years ago indicate that the skeletal elements are dispersed within the cave, but more recent data are not yet available. Most of the fossils are broken, with some breakage when the bone was fresh and some when already partly mineralized, both types showing some rounding. There are few longitudinal breaks on shafts of long bones and so very few bone splinters. All skeletal elements are preserved but in unequal proportions, with elements like femora, humeri and mandibles and teeth with greater structural density being best represented. There is no evidence of weathering or of human damage such as cut marks on any of the human assemblage, but trampling damage is present on most bones. Carnivore damage is also common, with some present on more than half the sample, but it is mostly superficial, either on the surfaces of shafts and articular ends or on the edges of spiral breaks. The sizes and distribution of the carnivore pits indicate extensive canid activity, and this is interpreted as scavenging of the bones in place in the cave. Indications of tooth marks from a larger carnivore indicate the activity possibly of a large felid: the marks are too large to be produced by small canids, with the larger marks concentrated on spiral breaks on the more robust bones, and there is no evidence of bone crushing and splintering in the manner of hyaenas. The nature of the SH human assemblage is also consistent with accumulation by humans, the evidence for this being the lack of other animals, especially the lack of herbivorous animals, associated with the humans, and the high number of individuals preserved.
Based on a survey of the literature on pretreatment of fused silica capillaries, 3 etching procedures and 11 silanization protocols based on the vinylic silane 3-((trimethoxysilyl)propyl) methacrylate (gamma-MAPS) were found to be most representative as a means of ensuring attachment of in situ prepared vinylic polymers. These techniques were applied to fused silica capillaries and the success in establishing the intended surface modification was assessed. X-ray photoelectron spectroscopy (XPS) was used to characterize the chemical state of the surface, providing information regarding presence of the reagent bound to the capillary. Wetting angles were measured and correlated with the XPS results. An adherence test was done by photopolymerization of a 2 mm long plug of 1,6-butanediol dimethacrylate in the prepared capillaries and evaluation of its ability to withstand applied hydraulic pressure. SEM was also performed in cases where the plug was released or other irregularities were observed. Finally, the roughness of the etched surface, considered to be of importance, was assessed by atomic force microscopy. Alkaline etching at elevated temperature provided a surface roughness promoting adhesion. The commonly used silanization protocols involving water in the silanization or washing steps gave inadequate surface treatment. The best silanization procedure was based on toluene as a solvent.
Caput and cauda epididymal fluids were found to be exceedingly rich in the numbers and kinds of polypeptides when analyzed by two-dimensional (2-D) gel electrophoresis. Only a few of the major (Coomassie-stained) and minor (silver-stained) epididymal fluid polypeptides were identified on epididymal sperm plasma membranes (PM) and even fewer identified in ejaculated sperm. The 2-D electrophoretic patterns of caput sperm PM differed little from those of cauda sperm PM. Thus, epididymal transit resulted in relatively minor quantitative and qualitative modifications in sperm PM composition. Seminal plasma showed a few major polypeptides from the cauda epididymal fluid, but the major constituents were those polypeptides from the seminal vesicle secretions. Sperm appear to acquire one acidic high molecular weight polypeptide from either the bulbourethral gland or prostate gland, and another major acidic polypeptide of high molecular weight from the seminal vesicle gland. Numerous neutral and basic low molecular weight polypeptides, originating from the seminal vesicles, adhered tightly to sperm. These were major polypeptides and constituted a substantial percentage of the total PM protein. Thus, major contributions to the sperm PM polypeptide profile occurred at ejaculation. This study did not address loosely bound polypeptides but is the first to analyze, in a comprehensive way, the origins of tightly bound sperm polypeptides from a single species.
Uniformly sized packing materials based on synthetic polymer particles for high-performance liquid chromatography (HPLC) and capillary electrochromatography (CEC) have been prepared from polymerization mixtures containing methacrylic acid (MAA) as a functional monomer and by using a novel surface modification method. This "dispersion method" affords effectively modified separation media. Both the amount of MAA utilized in the preparation and reaction time affect the selectivity of chromatographic separation in both the HPLC and the CEC mode and electroosmotic flow. This detailed study revealed that the dispersion method effectively modified internal surface of macroporous separation media and, based on the amount of MAA introduced, exclusion mechanism for the separation of certain solutes could be observed.
Rapid resealing of the mucosal epithelia is imperative following injuries to the small intestine because the mucosa is responsible for the adsorption of nutrients as well as providing a barrier to noxious agents present in the lumen. Tissue engineering may provide a possible solution for treating intestinal erosions, ulcerations, inflammatory bowel disease, and infection. Cell-biomaterial interaction is a critical component in tissue engineering that can determine the success of the tissue construct. Cell-biomaterial interactions can be enhanced by various types of surface modification, which promote integrin ligation leading to increased cell function. In order to relate the effect of surface adhesion molecules to signaling events and macroscopic cell response, an intestinal epithelial cell line, IEC-6, was plated on fibronectin (receptor-mediated) and poly-L-lysine (non-specific) surfaces. Focal adhesion kinase (FAK) phosphorylation, cell spreading, and cell adhesion strength were measured. Results showed increases in FAK phosphorylation generally corresponded to increases in cell spreading and adhesion strength for IEC-6 cells. Therefore, in a simplified system, initial adhesion and signaling mechanisms appeared to correspond to subsequent physical responses in IEC-6 cells relevant to tissue engineering applications.
In contrast to the high standard of intraocular lens implantation today, lens fixation and the occurrence of secondary opacifications are still not satisfactory. We are working on an implant that builds a stable connection to the surrounding lens-capsule. Disc-shaped lenses made of poly(dimethylsiloxane) were treated with different kinds of plasma to bring functional groups to the surface. This allows further macromolecular coating. The results of the surface modifications were characterized by electron spectroscopy for chemical analysis, scanning electron microscopy, Fourier transform infrared spectroscopy and cell culture experiments. O2 plasma induces a rapid increase of functional groups on the lens surface. CO2 plasma has to be used for longer treatment periods to achieve the same increase, but as SEM shows, did not lead to surface damage up to periods of 300 s. Using O2 plasma for longer than 60 s, a crack formation occurred. Therefore, CO2 plasma may be a more effective and sensitive tool in grafting functional groups to the surface.
Previous studies have shown that the carboxyl groups in hyaluronic acid (HA) could be activated by 1-ethy 1-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) to form intermediate O-acylisoureas, which can chemically bind to exposed amino groups on the tendon surface, leading to improved gliding ability. However, the optimal ratio and concentrations of the components in this chemical mixture were not investigated. The purpose of this study was to optimize the constituents of this tissue engineering approach to tendon surface modification, to reduce friction and improve durability. Peroneus longus (PL) tendons (n=40) were harvested from adult mongrel dogs along with the A2 pulley obtained from the ipsilateral hind paw. After the gliding resistance of the normal PL tendon was measured, the tendons were treated under varying concentrations of HA (0.5, 1, and 2%) and EDC/NHS (0.05, 0.25, and 1%) mixed with a 10% gelatin. Tendon friction was measured for 1000 cycles of simulated flexion/extension motion. Following testing, the residual HA on the tendon surface was evaluated by immunohistochemisty. The gliding resistance of the untreated PL tendons had a mean value of 0.087+/-0.021 N. After surface treatment, there was no significant difference in friction due to HA concentration alone, but the concentration of EDC/NHS and the interaction between HA concentration and EDC/NHS concentration had a significant effect on friction. Regardless of HA concentration, the friction after 1000 cycles was significantly decreased in preparations which included a 1% concentration of EDC/NHS. The tendons with lower gliding resistance presented a smoother surface on light microscopy and maintained more residual HA on the tendon surface. By varying the relative concentrations of HA, EDC, and NHS it is possible to optimize the effect of surface treatment on friction and durability in a canine extrasynovial tendon in vitro.
Tumor promotion activity of polyethylene (PE) was estimated in terms of the inhibitory potentials on the gap-junctional intercellular communication using the metabolic cooperation assay. The gap-junctional intercellular communication of test cells was inhibited on the PE film, but this inhibitory activity was markedly decreased when the surface of the PE film was immobilized with collagen. These results suggest that the in vivo tumor promotion activity of the untreated PE may be stronger than that of collagen-immobilized PE. On the other hand, surface modification with RGDS peptide, which is well known as the sequence of cell attachment domain in extracellular matrix proteins, did not reduce the promotion activity of PE film. In addition, neither modification with bovine serum albumin nor RGES peptide reduced the activity of PE film. These findings suggest that reduction of the inhibitory activity on gap-junctional intercellular communication by collagen immobilization is not simply due to improved cell adherence via the RGDS sequence but to some cell-cell recognition via collagen molecules essential for the gap-junctional intercellular communication.
Ultra-thin polycaprolactone (PCL) produced by bi-axial stretching was previously shown to have significant advantage for membrane tissue engineering. However, the permeability of the membrane needs to be enhanced. In this study, ablation experiments using femtosecond laser and excimer laser were carried out to modify the PCL surface. The use of the femtosecond laser produces neat drilled-through holes while the excimer laser is employed to produce blind-holes on the membrane. The modified surface of the membrane was studied and analyzed for different laser parameters (such as pulse energy and pulse repetition rate and characterized using several techniques that include optical microscopy, scanning electron microscopy and water contact angle measurements). Results showed that the morphological surface changes with different laser parameters, and the water contact angle decreases as the surface of the membrane is modified. The decrease in water contact angle suggests that surface of the membrane had become more hydrophilic than the non-laser treated membrane. The present study demonstrated that laser surface modification on the PCL can be achieved with high degree of success and precision. This paved the way for further enhancement in membrane tissue engineering.
BACKGROUND: Endovascular therapy with Guglielmi detachable coils is an accepted treatment option for patients with intracranial aneurysms. However, an emerging technology in the realm of endovascular tools is the use of traditional Guglielmi detachable coils with biologically active substances complexed to the coil surface to enhance aneurysm occlusion. METHODS: We review the literature and current trends in modified Guglielmi detachable coils. Surface modifications with extracellular matrix proteins, growth factors, ion impregnation, and genetically altered cells have been used in animal studies to improve the cellular response of Guglielmi detachable coils. Similarly, coronary artery stents have been modified in several different ways to maintain vessel patency, contrary to the goal of endovascular therapy. We comparatively reviewed this literature to add insight into the evolution of the research on modified Guglielmi detachable coils. CONCLUSIONS: Guglielmi detachable coil modifications have the potential to enhance aneurysm obliteration with directed cellular responses. This may allow aneurysm occlusion with coils in less time than untreated coils, thus decreasing the risks of aneurysm enlargement and hemorrhage.
BACKGROUND: Pathophysiology of extracorporeal circulation is multifactorial, and the link between newly developed "biomaterials" and clinical outcome is not easy to illustrate. MATERIAL AND METHODS: We designed a randomized, double-blinded, prospective study in order to verify the impact of a new surface modification [SMAR(X)T] in combination with an optimized blood air interface, on the cerebral performance after cardiopulmonary bypass. 80 patients were randomly divided into two subgroups (SMAR(X)T vs. standard PVC control) and analyzed for the kinetic of cerebral ischemia markers neuronspecific enolase, protein S100 and neuropsychologically tested with the Mini-Mental-Status Test (MMST) before and after the operation. RESULTS: We could not show significant differences of protein S100 and neuron-specific enolase (NSE) levels between SMAR(X)T patients and the controls, but the incidence of neurological complications was exceptionally low. Although no statistically significant differences could be obtained for the MMST, the different pointloss between both groups trends toward a better cerebral performance in SMAR(X)T patients. CONCLUSION: The use of a biologically inert circuit in combination with an optimized perfusion management seems to be worthy of recommendation.
With the growing general concern about the pollution by fly ash (FA), there has been global interest in its utilization. Purified FA or FA micro-beads are suitable as polymer filling materials because of their density, good dispersity and fluidity of globular particles. However, FA as a filler has not been widely used up to now on account of low whiteness values and low friction of untreated FA surface. In order to improve the FA quality, a surface modification method by using isothermal heating is proposed in this paper. Preparation of composite fly ash (CFA) in the Ca(OH)(2)-H(2)O-CO(2) system is described. Good coating results on FA surfaces can be achieved under suitable operating parameters. The characteristics of CFA are discussed and analyzed based on data from X-ray diffraction, scanning electron microscopy (SEM), infrared spectra, and BET multiple-point nitrogen adsorption method. Feedstocks with less than 45 microm grain size, 2.86 m(2) g(-1) specific surface area, and 36.68 whiteness value revealed an increase in specific surface area ranging from 8.69 to 10.01 m(2) g(-1) and an increase in whiteness values ranging from 63.67 to 73.13 after coating. A SEM study allowed a detailed determination of the morphology of the surface roughness. Filling tests also show that a rough surface of the CFA enhances contact opportunities and improves the interface between polymer and CFA blended with polypropylene (PP).
Poly(oxyethylene)-poly(oxypropylene) (PEO-PPO) co-polymers have been used as surfactants to produce resorbable poly(DL-lactide co-glycolide) (PLG) microspheres in the 500 nm-1 micron size range by an emulsification/solvent evaporation technique based on acetone-dichloromethane mixtures. The high polydispersity of microspheres could be reduced by using low PLG concentrations of 1% (w/v). Surface analysis by static secondary ion mass spectroscopy revealed the presence of PEO-PPO at the microsphere surface after cleaning by centrifuging and resuspension in water, and after further cleaning by dialysis. Physical entrapment of PEO-PPO chains in the particle surface is indicated due to rapid collapse of the solvent swollen PLG network as acetone is extracted from the suspended droplets. Opportunities are presented for simultaneous manufacture and surface modification of microspheres.
Restenosis is the reobstruction of an artery following interventional procedures such as balloon angioplasty or stenting. Local pharmacotherapeutic approaches using controlled release systems are under investigation to inhibit the regional pathophysiologic process of restenosis. We have been investigating biodegradable nanoparticles (100 +/- 39 nm in diameter, mean +/- sd) for the local intra-arterial drug delivery. The purpose of this study was to investigate nanoparticle surface modifications (see Table 1) to enhance their arterial uptake. The PLGA (polylactic polyglycolic acid copolymer) nanoparticles were formulated by an oil-in-water emulsion solvent evaporation technique using a 2-aminochromone (U-86983, Upjohn and Pharmacia) (U-86) as a model antiproliferative agent. The various formulations of nanoparticles were evaluated for the arterial wall uptake by using an ex-vivo dog femoral artery model. The selected formulations were then tested in vivo in acute dog femoral artery and pig coronary artery models. The nanoparticles surface modified with a cationic compound, didodecyldimethylammonium bromide (DMAB), demonstrated 7-10-fold greater arterial U-86 levels compared to the unmodified nanoparticles in different ex-vivo and in-vivo studies. The mean U-86 levels were 10.7 +/- 1.7 microg/10 mg (dog) and 6.6 +/- 0.6 microg/10 mg (pig) in the artery segments ( approximately 2 cm) which were infused with the nanoparticles. The pig coronary studies further demonstrated that the infusion of nanoparticles with higher U-86 loading reduced the arterial U-86 levels, whereas increasing the nanoparticle concentration in the infusion solutions increased the arterial U-86 levels. The biodistribution studies in pigs following coronary arterial administration of nanoparticles demonstrated disposition of U-86 in the myocardium and distally in the liver and the lung. The mechanism of enhanced arterial uptake of the DMAB surface modified nanoparticles seems to be due to the alteration in the nanoparticle surface charge. The unmodified nanoparticles had a zeta potential of -27.8 +/- 0.5 mV (mean +/- sem, n = 5), whereas the DMAB modified nanoparticles demonstrated a zeta potential of +22.1 +/- 3.2 mV (mean +/- sem, n = 5). The adsorption of DMAB to the nanoparticle surface followed the Freundlich isotherm with binding capacity k = 28.1 microg/mg and affinity constant p = 2. 33. In conclusion, surface modified nanoparticles have potential applications for intra-arterial drug delivery to localize therapeutic agents in the arterial wall to inhibit restenosis.