The endothelization of polyhedral oligomeric silsesquioxane nanocomposites

It has been recognized that seeding vascular bypass grafts with endothelial cells is the ideal method of improving their long-term patency rates. The aim of this study was to assess the in vitro cytocompatibility of a novel silica nanocomposite, polyhedral oligomeric silsesquioxane-poly(carbonate-urea)urethane (POSS-PCU) and hence elicit its feasibility at the vascular interface for potential use in cardiovascular devices such as vascular grafts. Using primary human umbilical vein endothelial cells (HUVEC), cell viability and adhesion were studied using AlamarBlue assays, whereas cell proliferation on the polymer was assessed using the PicoGreen dye assay. Cellular confluence and morphology on the nanocomposite were analyzed using light and electron microscopy, respectively. Our results showed that there was no significant difference between cell viability in standard culture media and POSS-PCU. Endothelial cells were capable of adhering to the polymer within 30 min of contact (Student's t-test, p<0.05) with no difference between POSS-PCU and control cell culture plates. POSS-PCU was also capable of sustaining good cell proliferation for up to 14d even from low seeding densities (1.0×10³ cells/cm²) and reaching saturation by 21 d. Microscopic analysis showed evidence of optimal endothelial cell adsorption morphology with the absence of impaired motility and morphogenesis. In conclusion, these results support the application of POSS-PCU as a suitable biomaterial scaffold in bio-hybrid vascular prostheses and biomedical devices.     

Polyhedral oligomeric silsesquioxane (POSS) suppresses enzymatic degradation of PCL-based polyurethanes

In this Article, we studied the enzymatic hydrolytic biodegradation behavior of a novel multiblock thermoplastic polyurethane (TPU) system, which incorporates polyhedral oligomeric silsesquioxane (POSS) into linear biodegradable thermoplastic polyurethanes containing poly(ε-caproactone) (PCL) and polyethylene glycol (PEG) blocks. The biodegradation behavior of POSS-PCL-PEG TPUs was characterized by proton nuclear magnetic resonance spectroscopy ((1)H NMR), differential scanning calorimetry (DSC), tensile tests, scanning electron microscopy (SEM), and wavelength dispersive X-ray spectrometry (WDS) after enduring 22-day accelerated enzymatic hydrolytic degradation tests. POSS incorporation significantly suppressed in vitro enzymatic hydrolytic degradation of PCL-PEG-based multiblock TPUs by a surface passivation mechanism. WDS observations revealed that the covalently bonded POSS moieties developed a near-continuous and robust POSS-layer after initial degradation, which prevented ester bonds of PCL from enzymatic attack, thereby inhibiting further degradation. These striking results provide a new strategy to fabricate the polyester-based biostable thermoplastic polyurethanes (TPUs) of potential use in long-term surgical implants.     

Structure of copper(II) complexes adsorbed on functionalized polyhedral oligomeric silsesquioxane

The 3-amino-1,2,4-triazole-modified silsesquioxane nanoplatforms have been prepared and characterized. The silsesquioxane nanocages readily react with CuX₂ in aqueous solution to form copper complex-substituted silsesquioxanes. Adsorption isotherms of CuX₂ from aqueous solution were studied at 25 °C. The electronic and ESR spectral parameters indicated that the copper ion is in a distorted-tetragonal symmetry field.     

The influence of porosity on the hemocompatibility of polyhedral oligomeric silsesquioxane poly (caprolactone-urea) urethane

BackgroundThe physio-chemical properties of blood contacting biomaterials play an important role in determining their hemocompatibility. It is shown in literature that surface roughness and porosity have significant effect on hemocompatibility. In this study, we use a biocompatible, low thrombogenic nanocomposite polymer called POSS-PCU to test this hypothesis: would porosity compromise the hemocompatibility of POSS-PCU. We compared the hemocompatibility of POSS-PCU films of various pore sizes with PTFE, which is a commercially available material used in most blood contacting devices.MethodsSterilized POSS-PCU films with different size pores were prepared as samples and porous PTFE film were selected as control. And all samples were subjected to SEM for topograpgy, mechanical test for characterization and hemocompatibility tests to evaluate contact activation, platelet adhesion and activation, as well as whole blood clotting response to the samples.ResultsWCA significantly increased with the pore size of POSS-PCU film, whereas both tensile stress and strain decreased significantly as the sizes of pores increased. However, when compared to PTFE film with same size pores, POSS-PCU films showed both higher tensile stress and strain. Pore size had little impact over POSS-PCU's surface chemistry groups as tested by FTIR analysis. Contact activation and platelet adhesion essay also showed no significant difference between different POSS-PCU samples. However, in whole blood reactions, POSS-PCU with pores size around 2–5 μm showed higher BCI than plain films and those with pores size around 35–45 μm. POSS-PCU showed lower thrombogencity and higher hemocompatibility comparing with porous PTFE on the aspects of platelet activation, adhesion and whole blood reaction.Summary and conclusionsPOSS-PCU polymer films as a biomaterial in chronic blood contacting implants show significant lower thrombogencity and higher hemocompatibility than porous PTFE film. It is desirable as a coating or covering material in small diameter stents for treating cardiovascular diseases, cerebral vascular diseases and peripheral arterial diseases.     

Preparation and characterization of cellulose hybrids grafted with the polyhedral oligomeric silsesquioxanes (POSS)

The cellulose hybrids with polyhedral oligomeric silsesquioxane (POSS) are synthesized by cross-linking graft reaction. Dimethylol dihydroxy ethylene urea (DDEU) as cross-linking agent is used in the graft reaction. The chemical and surface morphological structures of the am-POSS grafted cellulose hybrids are characterized with micro-FT-IR spectra, silicon element analysis, X-ray diffraction, SEM, AFM, and DSC. The results show that the am-POSS grafted cellulose hybrids form new macromolecular structures containing POSS nano-silica particles. POSS particles are evenly dispersed at the nanometer scale in the cellulose host matrix, bonding to the cellulose through covalent bonds. The thermal properties of the am-POSS grafted cellulose hybrids are improved.     

Enzymatic synthesis of a catechin conjugate of polyhedral oligomeric silsesquioxane and evaluation of its antioxidant activity

The antioxidant activity of catechin was amplified by conjugation with amine-terminated polyhedral oligomeric silsesquioxane (POSS) using horseradish peroxidase as catalyst. Compared to intact catechin, the scavenging activity of the POSS-catechin conjugate against superoxide anion was greatly improved. In addition, the conjugate strongly inhibited xanthine oxidase activity.     

Detection of DNA using cationic polyhedral oligomeric silsesquioxane nanoparticles as the probe by resonance light scattering technique

A novel cationic polyhedral oligomeric silsesquioxane nanoparticle (cationic POSS) was synthesized and successfully used as a new probe for the detection of DNA by resonance light scattering technique (RLS). It was found that the electrostatic interaction of cationic POSS and DNA could obviously enhance the RLS signal, the enhanced RLS intensity at 360 nm was proportional to the concentration of nucleic acids within the range of 0.35-42.82 microg ml-1 for calf thymus DNA, the determination limit (3sigma) was 0.32 ng ml-1. The results showed this method was very sensitive, convenient, rapid and reproducible.     

Development of a new lacrimal drainage conduit using POSS nanocomposite

Lacrimal surgery in cases of severely obstructed or missing canalicular ducts is highly challenging. In these cases, the placement of a bypass tube is currently the only option to restore the drainage of tears into the nose and reduce the symptomatic watery eye. Different approaches to achieve functional drainage have been tried using blood vessels or artificial implants. The implantation of the rigid Lester Jones tube is, since its introduction in the late 1960s, the gold standard. The functional success is satisfactory. However, complication rates are high and remain, even with many modifications of the original design, a major problem. These complications include mainly the displacement and blockage of the tube, requiring regular checkups, as well as irritation of the surrounding tissue including the nose and the eye. The objective of this study was to develop a new lacrimal duct conduit (LDC) to restore structural and functional integrity of the lacrimal drainage system. The conduit is constructed with a novel polymer, polyhedral oligomeric silsesquioxane-poly(carbonate-urea)urethane (POSS-PCU), that offers biocompatibility. We exploit nanotopography to evade the problems associated with current applications. A number of extrusion techniques were investigated for this purpose: ultrasonic atomization spraying, electrohydrodynamic atomization spraying/spinning, extrusion-coagulation, and high-pressure coagulation by autoclave and casting. Finally, the coagulation and cast technique were selected to construct an LDC superior to its predecessors, and its advantages highlighted.     

The oligomeric state sets GABA(B) receptor signalling efficacy

G protein-coupled receptors (GPCRs) have key roles in cell-cell communication. Recent data suggest that these receptors can form large complexes, a possibility expected to expand the complexity of this regulatory system. Among the brain GPCRs, the heterodimeric GABA(B) receptor is one of the most abundant, being distributed in most brain regions, on either pre- or post-synaptic elements. Here, using specific antibodies labelled with time-resolved FRET compatible fluorophores, we provide evidence that the heterodimeric GABA(B) receptor can form higher-ordered oligomers in the brain, as suggested by the close proximity of the GABA(B1) subunits. Destabilizing the oligomers using a competitor or a GABA(B1) mutant revealed different G protein coupling efficiencies depending on the oligomeric state of the receptor. By examining, in heterologous system, the G protein coupling properties of such GABA(B) receptor oligomers composed of a wild-type and a non-functional mutant heterodimer, we provide evidence for a negative functional cooperativity between the GABA(B) heterodimers.     

Preparation and antithrombogenic activities of heparinoid from 6-O-(carboxymethyl)chitin

The highest antithrombogenic activity was achieved by the sulphation of partially N-deacetylated O-(carboxymethyl)chitin among variously modified chitin derivatives. It was also suggested that the distribution of N-sulphate and N-acetyl groups on the C-2 position might be essential to the selective binding by antithrombin-III to inhibit thrombin activity. Kinetic evaluations demonstrated the non-competitive inhibition on direct interaction with thrombin (K_i = 9.26 × 10⁻⁸m) and the competitive inhibition with antithrombin-III (K_i = 3.33 × 10⁻⁸) m as well as with heparin. 6-O-Carboxymethyl groups were found, from the data of intravenous injection in mice, to suppress the toxicity of chitin heparinoids.     

The oligomeric Rep protein of Mungbean yellow mosaic India virus (MYMIV) is a likely replicative helicase

Geminiviruses replicate by rolling circle mode of replication (RCR) and the viral Rep protein initiates RCR by the site-specific nicking at a conserved nonamer (TAATATT downward arrow AC) sequence. The mechanism of subsequent steps of the replication process, e.g. helicase activity to drive fork-elongation, etc. has largely remained obscure. Here we show that Rep of a geminivirus, namely, Mungbean yellow mosaic India virus (MYMIV), acts as a replicative helicase. The Rep-helicase, requiring > or =6 nt space for its efficient activity, translocates in the 3'-->5' direction, and the presence of forked junction in the substrate does not influence the activity to any great extent. Rep forms a large oligomeric complex and the helicase activity is dependent on the oligomeric conformation ( approximately 24mer). The role of Rep as a replicative helicase has been demonstrated through ex vivo studies in Saccharomyces cerevisiae and in planta analyses in Nicotiana tabacum. We also establish that such helicase activity is not confined to the MYMIV system alone, but is also true with at least two other begomoviruses, viz., Mungbean yellow mosaic virus (MYMV) and Indian cassava mosaic virus (ICMV).     

3,N(4)-ethano-2'-deoxycytidine: chemistry of incorporation into oligomeric DNA and reassessment of miscoding potential

3,N(4)-Ethano-2'-deoxycytidine (ethano-dC) may be incorporated successfully into synthetic oligodeoxynucleotides by omitting the capping procedure used in the automated DNA synthetic protocols immediately after inserting the lesion and in all iterations thereafter. Ethano-dC is sensitive to acetic anhydride found in the capping reagent, and multiple oligomeric products are formed. These products were identified by examining the reaction of ethano-dC with the capping reagent, and several acetylated, ring-opened products were characterized by electrospray mass spectrometry and collision induced dissociation experiments on a tandem quadrupole mass spectrometer. A scheme for the formation of the acetylated products is proposed. In addition, the mutagenic profile of ethano-dC was re-examined and compared to that for etheno-dC. Ethano-dC is principally a blocking lesion; however, when encountered by the exo(-)Klenow fragment of DNA polymerase, dAMP (22%), TMP (16%), dGMP (5.3%) and dCMP (1.2%) were all incorporated opposite ethano-dC, along with an oligomer containing a one-base deletion (0.6%).     

Equilibrium unfolding of an oligomeric protein involves formation of a multimeric intermediate state(s)

Superoxide dismutases (SODs) are important metalloenzymes which protect cells against oxidative stress by scavenging reactive superoxides. Missense mutations in SODs are known to lead to some familial cases of amyotrophic lateral sclerosis and several forms of cancers. In the present study, we investigate the guanidinium hydrochloride (GdnHCl)-induced equilibrium unfolding of apo-manganese superoxide dismutase (apo-MnSOD) isolated from Vibrio alginolyticus using a variety of biophysical techniques. GdnHCl-induced equilibrium unfolding of apo-MnSOD is non-cooperative and involves the accumulation of stable intermediate state(s). Results of 1-anilino-8-naphthalene sulfonate binding experiments suggest that the equilibrium intermediate state(s) accumulates maximally in 1.5 M GdnHCl. The intermediate state(s) appears to be obligatory and occurs both in the unfolding and refolding pathways. Size-exclusion chromatography and sedimentation velocity data reveal that the equilibrium intermediate state(s) is multimeric. To our knowledge, this is the first report of the identification of a multimeric intermediate in the unfolding pathway(s) of oligomeric proteins. The formation and dissociation of the multimeric intermediate state(s) appears to dictate the fate of the protein either to refold to its native conformation or misfold and form aggregates as observed in amyotrophic lateral sclerosis.     

Flow behaviour of a POSS biopolymer solution

A non-biodegradable polyhedral oligomeric silsesquioxane (POSS) nanocomposite biopolymer has been developed for fabrication of medical devices and for tissue engineering human organs. The polymer in solution, containing 2 wt% of POSS, has been synthesized, characterized and investigated to determine its key rheological properties. Thus, the variation of shear stress and viscosity as a function of shear rate has been determined at ambient temperature to estimate yield stress and the index of pseudoplasticity, respectively. The temperature dependence of viscosity and the effect of ageing on the viscosity of the polymer have also been investigated. Results are compared with those of a conventional polycarbonate urethane (PCU) polymer solution. The POSS-PCU polymer solution shows near-Newtonian behaviour in the shear rate range to 1000 s(-1), having an apparent viscosity of approximately 3000 mPa s and a pseudoplasticity index of 0.90, decreasing slightly as the polymer solution is aged over 9 months. The temperature dependence of viscosity of the POSS polymer is extremely low and does not change with ageing but the yield strength increases from 2.7 Pa to 8.3 Pa.