Maternal-placental-fetal biodistribution of multimodal polymeric nanoparticles in a pregnant rat model in mid and late gestation.

Multimodal polymeric nanoparticles have many exciting diagnostic and therapeutic applications, yet their uptake and passage by the placenta, and applications in the treatment of pregnancy complications have not been thoroughly investigated. In this work, the maternal-fetal-placental biodistribution of anionic and cationic multimodal poly(glycidyl methacrylate) (PGMA) nanoparticles in pregnant rats at mid (ED10) and late (ED20) gestation was examined. Fluorescently-labelled and superparamagnetic PGMA nanoparticles functionalized with/without poly(ethyleneimine) (PEI) were administered to pregnant rats at a clinically-relevant dose and biodistribution and tissue uptake assessed. Quantitative measurement of fluorescence intensity or magnetic resonance relaxometry in tissue homogenates lacked the sensitivity to quantify tissue uptake. Confocal microscopy, however, identified uptake by maternal organs and the decidua (ectoplacental cone) and trophoblast giant cells of conceptuses at ED10. At ED20, preferential accumulation of cationic vs. anionic nanoparticles was observed in the placenta, with PGMA-PEI nanoparticles localised mainly within the chorionic plate. These findings highlight the significant impact of surface charge and gestational age in the biodistribution of nanoparticles in pregnancy, and demonstrate the importance of using highly sensitive measurement techniques to evaluate nanomaterial biodistribution and maternal-fetal exposure.

Experimental assessment of the effects of sublethal salinities on growth performance and stress in cultured tra catfish (Pangasianodon hypophthalmus).

The effects of a range of different sublethal salinities were assessed on physiological processes and growth performance in the freshwater 'tra' catfish (Pangasianodon hypophthalmus) juveniles over an 8-week experiment. Fish were distributed randomly among 6 salinity treatments [2, 6, 10, 14 and 18 g/L of salinity and a control (0 g/L)] with a subsequent 13-day period of acclimation. Low salinity conditions from 2 to 10 g/L provided optimal conditions with high survival and good growth performance, while 0 g/L and salinities >14 g/L gave poorer survival rates (p < 0.05). Salinity levels from freshwater to 10 g/L did not have any negative effects on fish weight gain, daily weight gain, or specific growth rate. Food conversion ratio, however, was lowest in the control treatment (p < 0.05) and highest at the maximum salinities tested (18 g/L treatment). Cortisol levels were elevated in the 14 and 18 g/L treatments after 6 h and reached a peak after 24-h exposure, and this also led to increases in plasma glucose concentration. After 14 days, surviving fish in all treatments appeared to have acclimated to their respective conditions with cortisol levels remaining under 5 ng/mL with glucose concentrations stable. Tra catfish do not appear to be efficient osmoregulators when salinity levels exceed 10 g/L, and at raised salinity levels, growth performance is compromised. In general, results of this study confirm that providing culture environments in the Mekong River Basin do not exceed 10 g/L salinity and that cultured tra catfish can continue to perform well.

Liver hydrolysate assists in the recovery from physical fatigue in a mouse model.

It is reported that liver hydrolysate (LH) enhances liver function. However, the effects of LH on physical fatigue are unknown. The aim of this study was to investigate the effect of LH on alterations in locomotor activity and energy metabolism such as 5'-AMP-activated protein kinase (AMPK), glycogen content, and blood lactic acid, after forced walking. Adult male ddY mice were used. Locomotor activity, AMPK phosphorylation, and glycogen content in the liver and soleus muscle, as well as blood lactic acid were determined following LH treatment before and/or after forced walking. The locomotor activity significantly decreased after forced walking for 3 h. Two administrations of LH (30 or 100 mg/kg) significantly increased the locomotor activity, while a single administration either before or after forced walking did not show any specific effect. Administering LH twice activated AMPK in the liver and soleus muscle. Glycogen levels significantly decreased in both the liver and soleus muscle after forced walking, whereas the blood lactate level significantly increased. In contrast, administering LH twice increased muscle glycogen and decreased blood lactic acid. These findings indicate that LH produced an anti-fatigue effect and that this effect appears to involve the efficient glycogen utilization through activation of AMPK.

An in vitro release study of indomethacin from nanoparticles based on methyl methacrylate/glycidyl methacrylate copolymers.

Indomethacin was coupled onto some macromolecular nanostructures based on methyl methacrylate copolymers with glycidyl methacrylate and tested as a model drug. The polymeric matrices were synthesized by radical emulsion copolymerization with and without the presence of a continuous external magnetic field of 1500 Gs intensity. Mathematical analysis of the release data was performed using Higuchi, Peppas-Korsmeyer equations. NIR chemical imaging (NIR-CI) was used to provide information about the spatial distribution of the components in the studied nanostructures. This opportunity was used to visualize the spatial distribution of bioactive substances (indomethacin) into the polymeric matrix, as well as to evaluate the degree of chemical and/or physical heterogeneity of the bioactive samples. The release rate dependence on the synthesis conditions as well as on the chemical compositions of the tested polymeric systems, it was also evidenced.

Immobilization of type-I collagen and basic fibroblast growth factor (bFGF) onto poly (HEMA-co-MMA) hydrogel surface and its cytotoxicity study.

Type-I collagen and bFGF were immobilized onto the surface of poly (HEMA-co-MMA) hydrogel by grafting and coating methods to improve its cytotoxicity. The multi-layered structure of the biocompatible layer was confirmed by FTIR, AFM and static water contact angles. The layers were stable in body-like environment (pH 7.4). Human skin fibroblast cells (HSFC) were seeded onto Col/bFGF-poly (HEMA-co-MMA), Col-poly (HEMA-co-MMA) and poly (HEMA-co-MMA) films for 1, 3 and 5 day. MTT assay was performed to evaluate the extraction toxicity of the materials. Results showed that the cell attachment, proliferation and differentiation on Col/bFGF-poly (HEMA-co-MMA) film were higher than those of the control group, which indicated the improvement of cell-material interaction. The extraction toxicity of the modified materials was also lower than that of the unmodified group. The protein and bFGF immobilized poly (HEMA-co-MMA) hydrogel might hold great promise to be a biocompatible material.

Ultrastructural evaluation of in vitro mineralized calcium phosphate phase on surface phosphorylated poly(hydroxy ethyl methacrylate-co-methyl methacrylate).

The in vitro functionality of surface phosphorylated poly(hydroxy ethyl methacrylate-co-methyl methacrylate), poly(HEMA-co-MMA) to induce bioinspired mineralization of calcium phosphate phase is evaluated. The primary nucleation of calcium phosphate on the surface phosphorylated copolymer occurs within 3 days of immersion when immersed in 1.5x simulated body fluid and the degree of mineralization is proportional to the hydroxy ethyl methacrylate content in the copolymer. The calcium phosphate phase is identified as hydroxyapatite by X-Ray diffraction analysis. The transmission electron microscopic evaluation combined with selected area diffraction pattern and energy dispersive analysis exemplified that the primary nuclei of amorphous calcium phosphate transforms to crystalline needle like calcium rich apatite, within a period of 3 days immersion in simulated body fluid. The atomic force microscopic results corroborate the c-axis growth of the crystals within 3 days immersion in SBF.

Adsorption of human salivary mucin MG1 onto glow-discharge plasma treated acrylic resin surfaces.

It has been suggested that altering the surface properties of acrylic resin material may change the nature of the adsorbed pellicle affecting denture retention and microbial adherence. This study aimed at evaluating the adsorption of salivary high molecular-weight mucins, a major component of denture pellicle, onto modified acrylic resin surfaces. (Poly) methylmethacrylate specimens were treated by glow discharge plasma technique, using hydrophilic 2-Hydroxyethylmethacrylate monomer or oxygen (O(2)) gas and hydrophobic Hexamethyldisiloxane monomer, at different discharge powers. Acrylic samples were incubated with high-molecular weight mucin, MG1 purified from saliva, the adsorbed fractions were transferred to nitrocellulose membranes by slot-blot technique, stained by periodic acid-Schiff and colour intensities were analysed by a colour densitometer. Higher amounts of mucins were adsorbed on all the surfaces modified by glow-discharge plasma treatment. Within the limitations of this study, it was concluded that glow-discharge plasma altered the surfaces of acrylic resin denture base materials and significantly increased the adsorption of high molecular-weight mucins at varying levels depending on plasma parameters.

Probing the S100 protein family through genomic and functional analysis.

The EF-hand superfamily of calcium binding proteins includes the S100, calcium binding protein, and troponin subfamilies. This study represents a genome, structure, and expression analysis of the S100 protein family, in mouse, human, and rat. We confirm the high level of conservation between mammalian sequences but show that four members, including S100A12, are present only in the human genome. We describe three new members of the S100 family in the three species and their locations within the S100 genomic clusters and propose a revised nomenclature and phylogenetic relationship between members of the EF-hand superfamily. Two of the three new genes were induced in bone-marrow-derived macrophages activated with bacterial lipopolysaccharide, suggesting a role in inflammation. Normal human and murine tissue distribution profiles indicate that some members of the family are expressed in a specific manner, whereas others are more ubiquitous. Structure-function analysis of the chemotactic properties of murine S100A8 and human S100A12, particularly within the active hinge domain, suggests that the human protein is the functional homolog of the murine protein. Strong similarities between the promoter regions of human S100A12 and murine S100A8 support this possibility. This study provides insights into the possible processes of evolution of the EF-hand protein superfamily. Evolution of the S100 proteins appears to have occurred in a modular fashion, also seen in other protein families such as the C2H2-type zinc-finger family.

Tailoring the swelling pressure of degrading dextran hydroxyethyl methacrylate hydrogels.

Swelling pressure measurements were performed on degrading dextran hydroxyethyl methacrylate (dex-HEMA) hydrogels. In these networks, the cross-links are hydrolyzable carbonate ester bonds formed between methacrylate groups and dextran molecules. It is demonstrated that dex-HEMA gels made in the presence of a known amount of free dextran chains exhibit osmotic properties similar to those of partially degraded dex-HEMA gels. The swelling pressure, Pi(sw), of degrading dex-HEMA gels is controlled primarily by the cross-linked dex-HEMA polymer and the free dextran molecules, while the contribution of short poly-HEMA fragments (produced in the degradation process) is negligible. It is found that Pi(sw) only slightly changes during the first 15 days of degradation. Close to the end of the degradation process, however, a much faster increase in Pi(sw) is observed. The swelling pressure profile of these gels strongly depends on the concentration of the cross-linked dex-HEMA and its chemical composition (amount of HEMA groups per 100 glucose units).

Enhanced anaerobic degradation of polymeric azo compounds by Escherichia coli in the presence of low-molecular-weight redox mediators.

The effects of the redox mediator lawsone (2-hydroxy-1,4-naphthoquinone) on the ability of Escherichia coli to reduce anaerobically polymeric azo compounds were analysed. Two types of polymeric azo compounds were tested, that have been proposed as putative tools for the site-specific targeting of drugs to the colon. The first group of polymers consisted basically of linear chains of polymethacrylic acid or polymethylmethacrylate which were interrupted by subunits of 4,4'-bis(methacryloylamino)azobenzene. These polymers differed significantly in their hydrophilicity according to the relative proportion of polymethacrylic acid used for the polymerization procedure. The second group of polymers consisted of almost water-insoluble poly(ether-ester)azo polymers that were composed of 4-(6-hydroxyhexyl)oxy-phenylazobenzoate and 16-hydroxyhexadecanoate. The addition of lawsone to the anaerobically incubated cultures of E. coli resulted in a pronounced increase in the reduction rates of the water-soluble poly(methacrylate-co-4,4'-bis(methacryloylamino)azobenzene) and in a much smaller, but significant, increase in the reduction rates of the hydrophobic poly(ether-ester)azo polymers. An increase in the amount of azo groups resulted, for the hydrophobic poly(ether-ester)azo polymers, in an increased reduction rate in the presence of the redox mediator lawsone.

Biocompatibility of NDGA-polymerized collagen fibers. II. Attachment, proliferation, and migration of tendon fibroblasts in vitro.

The material properties of collagen fibers polymerized with nordihydroguaiaretic acid (NDGA) are equivalent to native tendon, suggesting that NDGA crosslinking may provide a viable approach to stabilizing collagenous materials for use in repairing ruptured, lacerated, or surgically transected fibrous tissues, such as tendons and ligaments (Koob & Hernandez, Biomaterials, in press). The present study evaluated the biocompatibility of these fibers with cultured bovine tendon fibroblasts. Fibroblast attachment, migration, and proliferation on NDGA-crosslinked materials were compared to those on prepolymerized type I tendon collagen constructs as well as on tissue-culture-treated plastic. Fibroblast attachment on NDGA-crosslinked collagen fibrils was equivalent to attachment on plates coated with collagen alone. Over a period of 8 days in culture, attached fibroblasts proliferated on NDGA-crosslinked collagen at a rate identical to that of fibroblasts attached to native collagen. In order for the biomaterial effectively to bridge gaps in fibrous tissues, fibroblasts must be able to migrate and replicate on the bridging fiber. Control and crosslinked fibers were inserted in calf tendon explants, with a portion of the fiber extending out of the sectioned end of the tendon. Explants were cultured for 9 weeks, and the number of cells was measured at weekly intervals. Cells appeared on the fibers after 1 week of culture. By 2 weeks, cells had colonized the entire fiber. The number of cells continued to increase throughout the 9 weeks in culture, forming a layer several cells thick. Histologic analysis indicated that the fibroblasts populating the fibers appeared to originate in the epitenon. There was no difference in the rate of fibroblast migration and replication, nor in the ultimate number of colonizing cells, between control collagen fibers and NDGA-crosslinked fibers. NDGA-crosslinked fibers may provide a means of bridging gaps in ruptured, lacerated, or surgically transected tendons by providing a mechanically competent scaffold on which tendon fibroblasts can migrate, attach, and proliferate.

A hydrophobic interaction site for lysozyme binding to polyethylene glycol and model contact lens polymers.

Proton nuclear magnetic resonance (1H-NMR) spectroscopy is used to identify a preferred binding site for uncharged hydrophilic polymers on the surface of hen egg-white lysozyme. Chemical shift titrations show that exchangeable proton signals from amino acids Arg-61, Trp-62, Trp-63, Arg-73, Lys-96 and Asp-101 are selectively perturbed upon binding of poly(ethylene oxide), poly(ethylene glycol) and poly(ethylene-co-propylene oxide). The greatest binding-induced chemical shift changes are observed for Trp-62, Arg-61 and Arg-73 at the edge of the active site cleft of the protein, consistent with a predominantly hydrophobic interaction mode involving the polymer ethylene moieties. The more hydrophilic species poly(dihydroxypropyl methacrylate) causes similar but substantially smaller chemical shift effects than the other polymers, confirming the nature of the interaction. A dissociation constant of 76+/-5 mM is determined for the poly(ethylene glycol)-lysozyme complex. The relatively low affinity of the protein-polymer interactions compared to oligosaccharide substrate binding suggests that lysozyme activity is minimally affected by these materials.

[Heat of polymerization of bone cement can induce cardiac arrest]. / Knoglecementens haerdningsvarme kan udløse hjertestop.

Total hip arthroplasty is associated with cardiopulmonary complications including cardiac arrest. We present one of four cases of cardiac arrest, two of the cases were fatal. The pathogenesis suggested to explain these complications is venous air embolism, generated by the methylmethacrylate bone cement polymerization causing thermal blood damage. To prevent this happening cortical bone allotransplantation around the prosthesis and bone cement with a low temperature of polymerization may be used.

Embedding of bone samples in methylmethacrylate: an improved method suitable for bone histomorphometry, histochemistry, and immunohistochemistry.

Methylmethacrylate (MMA) embedding of undecalcified bone biopsies is a technique widely used for bone histomorphometry. However, conventional MMA embedding causes almost complete loss of enzyme activity and protein antigenicity in the tissues. Recently, an MMA embedding technique has been reported that preserves enzyme activity and antigenic determinants in bone tissue. We describe here a modification of this embedding method. For our modified MMA embedding process, commercially available methacrylates can be used without purification, and the histologic quality of bone sections is comparable to that of conventionally MMA-embedded bone specimens. The technique reported here can be employed for embedding of larger bone samples and is suitable for histochemical and immunohistological applications as well as for routine bone histomorphometry. By addition of methylbenzoate during infiltration and polymerization of the plastic, the antigenicity of the tissue was improved. As applications of this novel technique, demonstration of alkaline phosphatase and tartrate-resistant acid phosphatase as well as positive labeling of Kupffer cells and osteoclasts with the monoclonal antibody ED1 in sections of liver, tibiae, and vertebrae of 3-month-old rats was demonstrated. The method described here might be useful for the inclusion of histochemical and immunohistological methods into bone histomorphometry.

The Microsponge Delivery System (MDS): a topical delivery system with reduced irritancy incorporating multiple triggering mechanisms for the release of actives.

The Microsponge Delivery System (MDS) is a unique technology for the controlled release of topical agents and consists of macroporous beads, typically 10-25 microns in diameter, loaded with active agent. When applied to the skin, the MDS releases its active ingredient on a time mode and also in response to other stimuli (rubbing, temperature, pH, etc). MDS technology is being used currently in cosmetics, over-the-counter (OTC) skin care, sunscreens and prescription products. By delivering the active gradually to the skin, MDS-benzoyl peroxide formulations, for example, have excellent efficacy with minimal irritation. These are typical benefits from the use of the MDS.

Protein A carrying monosize PMMA microbeads for the removal of HIgG from human plasma.

Protein A-incorporated polymethylmethacrylate (PMMA) microbeads were investigated for specific removal of HIgG from human plasma. The microbeads were prepared by a phase inversion polymerization, and activated by periodate oxidation. Protein A was then incorporated by covalent binding onto these microbeads through hydroxyl groups coming from the stabilizer. The amount of incorporated protein A was controlled by the initial concentrations of protein A in the immobilization medium and pH. The maximum protein A immobilization of 0.615 mg protein A/g PMMA, was observed at a pH of 9.5 corresponding to an initial protein A concentration of 0.1 mg/ml. There was no HIgG adsorption onto the plain PMMA microbeads, while high HIgG adsorptions of up to 32 mg HIgG/g PMMA were achieved with human plasma.

In vitro bacterial adherence to hydrogel and poly(methyl methacrylate) intraocular lenses.

PURPOSE: To compare the in vitro adherence of Staphylococcus epidermidis to poly-(hydroxyethyl methacrylate) (polyHEMA) or hydrogel intraocular lenses (IOLs) and poly(methyl methacrylate) (PMMA) IOLs. SETTING: Lions Eye Institute, Perth, Western Australia. METHODS: One-piece hydrogel lenses and one-piece PMMA lenses were suspended for 60 minutes in standardized suspensions of a well-characterized strain of S. epidermidis and then sonicated in a known quantity of balanced salt solution to remove the adherent bacteria. Quantitative cultures of the sonicates were performed and the results analyzed statistically. RESULTS: The mean bacterial adherence of S. epidermidis to the PMMA IOLs (58,400 CFU) was more than 20 times greater than that to the hydrogel IOLs (1953 CFU). The difference was statistically significant (P < .001). CONCLUSIONS: Adherence of S. epidermidis to hydrogel IOLs is significantly lower than to PMMA IOLs. This suggests that the risk of postoperative endophthalmitis after cataract extraction and IOL implantation may be lower with the use of hydrogel IOLs.

Annexin V-coated intraocular lenses.

PURPOSE: To assess whether annexin V-coated poly(methyl methacrylate) (PMMA) intraocular lenses (IOLs) prevent postoperative inflammation in rabbit eyes. SETTING: Department of Ophthalmology, Hôpital Purpan, Toulouse, France. METHODS: Thirteen IOLs coated with annexin V were implanted in 13 rabbit eyes; the contralateral eyes received uncoated IOLs. Postoperative fibrin was quantitated by daily slitlamp examination until the anterior chamber was completely clear. Results were analyzed using a Wilcoxon test. Ocular toxicity was evaluated by light and electron microscopy. RESULTS: Eyes with the annexin V-coated IOLs had less severe inflammation on the first postoperative day, and the inflammation resolved more quickly than in eyes with uncoated IOLs (P < .05). No annexin V was released postoperatively, nor were there signs of ocular toxicity. CONCLUSION: Annexin V-coated lenses effectively reduced postoperative inflammation in rabbit eyes.