Induction of an in vitro reversible hypometabolism through chitosan-based nanoparticles

Objective: Chitosan-based nanoparticles (NPs) were prepared to promote intracellular sustained delivery of the synthetic delta opioid D-Ala(2)-D-Leu(5)-enkephalin (DADLE), prolonging peptide activity and inducing a safe and reversible hypometabolic state.

Materials and methods: NPs were prepared by combining ionotropic gelation and ultrasonication treatment. NP uptake studies and the effects of encapsulated DADLE on HeLa cells proliferation were tested by transmission electron microscopy (TEM) analysis, by immuno-fluorescence and immuno-cytochemistry.

Results: DADLE-loaded NPs are produced with suitable characteristics, a satisfactory process yield (55.4%?±?2.4%) and encapsulation efficiency (64.6%?±?2.1%). NPs are effective in inducing a hypometabolic stasis at a 10^?4?M DADLE concentration. Moreover, as seen from the immunofluorescence study, the effect persists through the recovery period (72?h). Indeed, NPs labelled by anti-enkephalin antibody inside cell nucleus reassert that the in vivo release of the peptide can be prolonged with respect to the case of free peptide supply.

Conclusion: The nanoparticulate drug delivery system described seems to be effective in inducing and prolonging a sort of hibernation-like state in the cells.     

Drug delivery strategies using polysaccharidic gels

Hydrogels are hydrophilic polymeric networks, with chemical or physical crosslinks, that are capable of swell and can retain a large amount of water. Among the numerous types of macromolecules that can be used for hydrogel formation, polysaccharides show very attractive advantages in comparison to synthetic polymers. They are widely present in living organisms, are usually abundant and show a number of peculiar physicochemical properties; furthermore, these macromolecules are, in most cases, non-toxic, biocompatible and can be obtained from renewable sources. For these reasons, polysaccharides seem to be particularly suitable for different applications in the wide field of pharmaceutics. As examples of the studies that have been carried out on this topic, this review will focus on two polysaccharides, alginate and xyloglucan. Alginate has been, and still is, extensively investigated and has numerous industrial applications, whereas xyloglucan was chosen because, although it has been much less studied, it shows interesting properties that should find important practical uses in the near future. The possible advantages of physical gels over those that are chemically crosslinked are also discussed.     

Genipin-crosslinked polyvinyl alcohol/silk fibroin/nano-hydroxyapatite hydrogel for fabrication of artificial cornea scaffolds—a novel approach to corneal tissue engineering

Abstract

Design of artificial corneal scaffolds substitute is crucial for replacement of impaired cornea. In this paper, porous polyvinyl alcohol/silk fibroin/nano-hydroxyapatite (PVA/SF/n-HA) composite hydrogel was prepared via the genipin (GP) cross-linking, the pore diameter of the hydrogel ranged from 8.138?nm and 90.269?nm, and the physical and physiological function of hydrogel were investigated. The resulting hydrogel exhibited favourable physical properties. With the GP content increasing, the structural regularity of PVA/SF/n-HA composite hydrogel was enhanced and the thermal stability was improved. The moisture content was slightly decreased and generally maintained at approximately 70%. The tensile strength was heightened up to 0.64?MPa, while the breaking elongation was decreased slightly. Moreover, the biofunction was investigated. The in vitro degradation test demonstrated that with the addition of GP, the stability of the composite hydrogels in protease XIV solution was promoted and the three-dimensional porosity structure of composite hydrogels was maintained as ever. And the human corneal fibroblasts (HCFs) were employed to examine the cells cytotoxicity of the PVA/SF/n-HA composite hydrogels with different GP content by CCK-8 assay. Based on confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM), HCFs had individually commendable adhesion and proliferation on PVA/n-HA/SF composite hydrogel. HCFs proliferated and grew into the pores of composite hydrogel. The results of biocompatibility experiments of composite hydrogel suggested that it was no acute toxicity, in vitro cytotoxicity was 0 or 1 grade. Overall, results from this paper, PVA/n-HA/SF composite hydrogel was a qualified medical material which conformed to the national standard, could be a promising alternative for artificial cornea scaffold material—a novel approach to corneal tissue engineering.     

Anti‐erosive effects of fluoride and phytosphingosine: an in vitro study

A selection of commercially available products containing stannous fluoride (SnF₂)/sodium fluoride (NaF), SnF₂/amorphous calcium phosphate (ACP), SnF₂/NaF/ACP, tin (Sn)/fluorine (F)/chitosan were compared with phytosphingosine (PHS) with respect to their anti‐erosive properties in vitro. One‐hundred and twenty bovine enamel specimens were immersed in the respective product slurries for 2 min, twice daily. The formulations were diluted with either remineralization solution or artificial saliva. After each treatment, an erosive challenge was performed for 10 min, twice daily, using citric acid, pH 3.4. The specimens were stored in remineralization solution or artificial saliva until the next treatment‐erosion challenge. After 10 d, tissue loss was determined using profilometry. Enamel softening was determined through surface microhardness measurements. Tissue‐loss values (measured in μm and expressed as mean ± SD) for PHS, SnF₂/NaF, SnF₂/ACP, SnF₂/ACP/NaF, and Sn/F/chitosan treatment groups and for the negative‐control group, were, respectively, 35.6 ± 2.8, 15.8 ± 1.8, 22.1 ± 2.0, 22.9 ± 1.8, 16.2 ± 1.2, and 51.2 ± 4.4 in the presence of remineralization solution and 31.7 ± 3.3, 15.6 ± 2.9, 16.5 ± 2.7, 16.8 ± 2.1, 13.1 ± 3.0, and 50.7 ± 2.8 in the presence of artificial saliva. There were no significant differences in surface microhardness measurements between the treatment groups. In conclusion, PHS resulted in a significant reduction of tissue loss compared with the negative control, but in comparison, the toothpastes containing Sn²⁺ and F^? ions were significantly more effective compared with PHS.     

Novel electrochemical xanthine biosensor based on chitosan–polypyrrole–gold nanoparticles hybrid bio-nanocomposite platform

The aim of this study was the electrochemical detection of the adenosine-3-phosphate degradation product, xanthine, using a new xanthine biosensor based on a hybrid bio-nanocomposite platform which has been successfully employed in the evaluation of meat freshness. In the design of the amperometric xanthine biosensor, chitosan–polypyrrole–gold nanoparticles fabricated by an in situ chemical synthesis method on a glassy carbon electrode surface was used to enhance electron transfer and to provide good enzyme affinity. Electrochemical studies were carried out by the modified electrode with immobilized xanthine oxidase on it, after which the biosensor was tested to ascertain the optimization parameters. The Biosensor exhibited a very good linear range of 1–200 μM, low detection limit of 0.25 μM, average response time of 8 seconds, and was not prone to significant interference from uric acid, ascorbic acid, glucose, and sodium benzoate. The resulting bio-nanocomposite xanthine biosensor was tested with fish, beef, and chicken real-sample measurements.     

Ultrasound Phantom Using Sodium Alginate as a Gelling Agent

For medical workers, ultrasound phantoms for human soft tissue are used not only for accuracy management of ultrasound diagnosis but also to aid ultrasound‐guided needle and blind catheter insertion training without risk to real patients. For the phantoms, ultrasound characteristics and a texture are required to mimic the human soft tissue. The proposed phantom was composed of sodium alginate, calcium sulfate dihydrate, trisodium phosphate 12‐hydrate, glycerol, and water. The propagation speed, attenuation coefficient, acoustic impedance, and texture of the proposed phantom were almost the same as those of human soft tissue. Expensive chemicals and special equipment are not required.     

A temperature-induced and shear-reversible assembly of latanoprost-loaded amphiphilic chitosan colloids: Characterization and in vivo glaucoma treatment

Hydrogels composed of assembled colloids is a material class that is currently receiving much interest and shows great promise for use in biomedical applications. This emerging material class presents unique properties derived from the combination of nanosized domains in the form of colloidal particles with a continuous gel network and an interspersed liquid phase. Here we developed an amphiphilic chitosan-based, thermogelling, shear-reversible colloidal gel system for improved glaucoma treatment and addressed how preparation procedures and loading with the anti-glaucoma drug latanoprost and commonly used preservative benzalkonium chloride influenced the mechanical properties of and drug release from the colloidal gels. The results highlight that incorporated substances and preparation procedures have effects both on mechanical properties and drug release, but that the release of drug loaded in the colloidal carriers is mainly limited by transport out of the carriers, rather than by diffusion within the gel. The developed colloidal chitosan based gels hold outstanding biomedical potential, as confirmed by the ease of preparation and administration, low cytotoxicity in MTT assay, excellent biocompatibility and lowering of intraocular pressure for 40 days in a rabbit glaucoma model. The findings clearly justify further investigations towards clinical use in the treatment of glaucoma. Furthermore, the use of this shear-reversible colloidal gel could easily be extended to localized treatment of a number of critical conditions, from chronic disorders to cancer, potentially resulting in a number of new therapeutics with improved clinical performance.     

In vitro and in vivo evaluation of polymer hydrogels for hemorrhage control

In vitro and in vivo experimentation of various synthetic polymer hydrogels was conducted to establish some of the integral material properties that influence hemostasis. In vitro swelling experiments suggested that positive electrostatic charge was a key determinant of the ability of a polymer hydrogel to absorb physiological fluids, e.g. human plasma and blood. In vitro testing using unadulterated sheep blood suggested positive electrostatic charge and crosslink density were key determinants of the ability of a material to induce or enhance clot formation. Hydrogel formulations composed of higher amounts of positive electrostatic charge and lower crosslink density were able to effectively induce and enhance clot formation in the presence of a coagulation cascade activator. In vivo experimentation confirmed that hydrogels containing higher electrostatic charge and low crosslink density are more effective at fostering the formation of a robust hemostatic plug to control blood loss.     

Allan Hoffman: A 70th Birthday Celebration

Water sorption which is not classically Fickian has been observed in a variety of polymers. Deviation from Fickian kinetics is widely assumed to be caused by rate-limiting polymer relaxation, despite minimal proof of this. To the contrary, the evidence accumulated in this work indicates that water transport in initially glassy poly(2-hydroxyethyl methacrylate) (PHEMA), an important water-swellable biomedical polymer, is controlled by Fickian diffusion. First of all, the fractional water uptake is initially linear and independent of sample thickness when plotted against the square root of time over initial thickness, as expected for a Fickian process. Furthermore, the moving solvent front also advanced with the square root of time. Temperature, polymer thermal history and initial solvent concentration all affected the sorption kinetics of PHEMA in manners consistent with a Fickian process. The invariably Fickian sorption mechanism is believed to be the consequence of the water molecule's small size and affinity for hydrophilic, swellable polymers.