In a biological system, nanoparticles (NPs) may interact with biomolecules. Specifically, the adsorption of proteins on the nanoparticle surface may influence both the nanoparticles' and proteins' ...overall bio-reactivity. Nevertheless, our knowledge of the biocompatibility and risk of exposure to nanomaterials is limited. Here, in vitro and ex ovo biocompatibility of naturally based crosslinked freeze-dried 3D porous collagen/chitosan scaffolds, modified with thermostable fibroblast growth factor 2 (FGF2-STAB
), to enhance healing and selenium nanoparticles (SeNPs) to provide antibacterial activity, were evaluated. Biocompatibility and cytotoxicity were tested in vitro using normal human dermal fibroblasts (NHDF) with scaffolds and SeNPs and FGF2-STAB
solutions. Metabolic activity assays indicated an antagonistic effect of SeNPs and FGF2-STAB
at high concentrations of SeNPs. The half-maximal inhibitory concentration (IC50) of SeNPs for NHDF was 18.9 µg/ml and IC80 was 5.6 µg/ml. The angiogenic properties of the scaffolds were monitored ex ovo using a chick chorioallantoic membrane (CAM) assay and the cytotoxicity of SeNPs over IC80 value was confirmed. Furthermore, the positive effect of FGF2-STAB
at very low concentrations (0.01 µg/ml) on NHDF metabolic activity was observed. Based on detailed in vitro testing, the optimal concentrations of additives in the scaffolds were determined, specifically 1 µg/ml of FGF2-STAB
and 1 µg/ml of SeNPs. The scaffolds were further subjected to antimicrobial tests, where an increase in selenium concentration in the collagen/chitosan scaffolds increased the antibacterial activity. This work highlights the antimicrobial ability and biocompatibility of newly developed crosslinked collagen/chitosan scaffolds involving FGF2-STAB
and SeNPs. Moreover, we suggest that these sponges could be used as scaffolds for growing cells in systems with low mechanical loading in tissue engineering, especially in dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration. Due to their antimicrobial properties, these scaffolds are also highly promising for tissue replacement requiring the prevention of infection.
The current limitations of calcium phosphate cements (CPCs) used in the field of bone regeneration consist of their brittleness, low injectability, disintegration in body fluids and low ...biodegradability. Moreover, no method is currently available to measure the setting time of CPCs in correlation with the evolution of the setting reaction. The study proposes that it is possible to improve and tune the properties of CPCs via the addition of a thermosensitive, biodegradable, thixotropic copolymer based on poly(lactic acid), poly(glycolic acid) and poly(ethylene glycol) (PLGA⁻PEG⁻PLGA) which undergoes gelation under physiological conditions. The setting times of alpha-tricalcium phosphate (α-TCP) mixed with aqueous solutions of PLGA⁻PEG⁻PLGA determined by means of time-sweep curves revealed a lag phase during the dissolution of the α-TCP particles. The magnitude of the storage modulus at lag phase depends on the liquid to powder ratio, the copolymer concentration and temperature. A sharp increase in the storage modulus was observed at the time of the precipitation of calcium deficient hydroxyapatite (CDHA) crystals, representing the loss of paste workability. The PLGA⁻PEG⁻PLGA copolymer demonstrates the desired pseudoplastic rheological behaviour with a small decrease in shear stress and the rapid recovery of the viscous state once the shear is removed, thus preventing CPC phase separation and providing good cohesion. Preliminary cytocompatibility tests performed on human mesenchymal stem cells proved the suitability of the novel copolymer/α-TCP for the purposes of mini-invasive surgery.
Treatment of complete loss of skin thickness requires expensive cellular materials and limited skin grafts used as temporary coverage. This paper presents an acellular bilayer scaffold modified with ...polydopamine (PDA), which is designed to mimic a missing dermis and a basement membrane (BM). The alternate dermis is made from freeze-dried collagen and chitosan (Coll/Chit) or collagen and a calcium salt of oxidized cellulose (Coll/CaOC). Alternate BM is made from electrospun gelatin (Gel), polycaprolactone (PCL), and CaOC. Morphological and mechanical analyzes have shown that PDA significantly improved the elasticity and strength of collagen microfibrils, which favorably affected swelling capacity and porosity. PDA significantly supported and maintained metabolic activity, proliferation, and viability of the murine fibroblast cell lines. The in vivo experiment carried out in a domestic Large white pig model resulted in the expression of pro-inflammatory cytokines in the first 1-2 weeks, giving the idea that PDA and/or CaOC trigger the early stages of inflammation. Otherwise, in later stages, PDA caused a reduction in inflammation with the expression of the anti-inflammatory molecule IL10 and the transforming growth factor β (TGFβ1), which could support the formation of fibroblasts. Similarities in treatment with native porcine skin suggested that the bilayer can be used as an implant for full-thickness skin wounds and thus eliminate the use of skin grafts.
Different topical hemostatic materials are used to achieve effective hemostasis. High hemostatic activity, biocompatibility, bioresorbability, and easy manipulation are to be expected in such a ...developed product. In the surgical world with these specific requirements, finding a proper hemostatic agent is very difficult. The study compared several materials of various construction properties, which were assessed for structural and related properties by morphological analyses and assessed in vivo for their efficiency and behaviour using a model of rat partial nephrectomy. New sodium salt of carboxymethyl cellulose (CMC) sponge with the lowest porosity and free swell absorptive capacity contained the highest amount of hydroxyl and carboxyl groups. Results revealed that this CMC material in the form of a bioresorbable sponge may ensure the necessary hemostatic effects, while also providing a positive influence on the reaction of the local tissue. The CMC material also needed significantly less time to achieve hemostasis (
p
< 0.001). Moreover, the sponge reached satisfactory results in the histopathological evaluation with the lowest destruction score and favorable healing reaction. This modified product proved itself to be a promising bioresorbable hemostat, which, according to its design, matches with its surgical applications. In general, the obtained data elucidated the dependency of the total effect on its structure and composition.
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Polar and apolar moieties of humic acids are spatially separated forming domains of different polarity. In this work, we tested the procedures to crosslink functional groups in polar domains of humic ...acids by using carbodiimide coupling and analyzed to which extent influenced the modification their hydration properties and stability. For this reason, we prepared eight derivatives of lignite humic acids using either water-soluble N-Ethyl-N′-(3-dimethylaminopropyl)carbodiimide (EDC) or water-insoluble N,N′-dicyclohexylcarbodiimide (DCC) under various conditions. Characterization of prepared derivatives showed that both methods lead to formation of crosslinked humic structures. Using of EDC resulted in lower degree of crosslinking, but better hydration properties. Higher moisture uptake and water holding capacity were observed in humic acids, which were pre-wetted prior to crosslinking for at least 24h. Although the EDC derivatives of humic acids contained only between 60 and 85% of original free carboxylic groups, they showed similar moisture uptake as parental humic acids by equilibration at relative humidities of both 94% and 76%. Under water-saturated conditions, the EDC derivatives showed faster swelling kinetics and reached almost the same water holding capacity as the original sample after 18days. However, both the EDC and DCC derivatives began to degrade already after 3–9days during swelling tests, which subsequently decreased their hydration. The results suggested that water holding capacity, swelling kinetics and moisture uptake of humic acids were not influenced significantly by the amount of polar groups, but also by their spatial arrangement and distribution.
•Carbodiimide crosslinking improved structural rigidity and hydration of humic acids.•Under water-saturated conditions, all derivatives partly degraded within several days.•Hydration of humic acids depends on both distribution and content of polar groups.
The newest trends in wound healing management and the development of the next generation of dressings are pointing toward natural polymeric materials with important beneficial properties such as ...antimicrobial effects, renewability, easier process of preparation, and biological activity. Here, we present the preparation and in vitro evaluation of a unique biopolymeric blend composed of natural polymers based on the positively charged polysaccharide chitosan and negatively charged gum karaya. A plate lysis assay of gum karaya and chitosan solution mixtures proved the synergistic antimicrobial effect against specific strains of both Gram-positive and Gram-negative bacteria and yeast. This polymeric mixture was used for hydrogel film preparation and determination of the composition effect on physical properties (swelling behavior in different solvents, pH, diffusion mechanism, hydrolytic stability, mechanical and optical properties). While the pure gum karaya with poly(vinyl alcohol) exhibited the highest hydrolytic degradation (68%), the mixture of poly(vinyl alcohol) and gum karaya with chitosan (in the 25:75 ratio) exhibited the lowest degradation value (41%) due to the strong physical interactions. Cytotoxicity tests performed with hydrogel extracts using two different in vitro models, adherent fibroblasts (NIH3T3) and non-adherent suspension B-lymphocytes (BaF3), exhibited excellent biocompatibility and no cytotoxicity. As expected, the antimicrobial activity of 3-day film extracts showed a significantly improved antimicrobial effect of mixtures involving a chitosan biopolymer. The physical and biological properties of prepared biopolymer-based hydrogels meet the requirements of modern wound dressings.
The aim of this study was to answer the question whether our newly developed injectable biodegradable “self-setting” polymer-composite as a bone adhesive is a good “bone-glue” candidate to ...efficiently fix comminuted fractures of pig femoral bones used as an ex-vivo experimental model.
Mechanical properties of adhesive prepared from α-tricalcium phosphate (TCP) powder and thermogelling copolymer were optimized by selecting the appropriate composition with adhesion enhancers based on dopamine and sodium iodinate. Setting time and injectability were controlled by rheology. Ex-vivo experiments of fixed pig bones were provided in terms of either the three-point bending test of bending wedge type fractured pig femurs (with LCP) or the axial compression test of 45° oblique fractured femurs (without LCP) in physiological saline solution at 37 °C. Fractured bones treated with optimized adhesive before and after bending tests were imaged by X-ray microtomography (μCT).
Based on the rheological measurement, the adhesive modified with both dopamine and sodium iodinate exhibited optimal thixotropic properties required for injection via thin 22 G needle. This optimal adhesive composition showed an 8 min lag phase (processing time) followed by fast increase in storage modulus at 37 °C up to 1 GPa within 110 min. Self-setting of dopamine/iodinate modified adhesive was completed in 48 h exhibiting the maximum strength at compression of 7.98 MPa ± 1.39 MPa. Whereas unmodified adhesive failed in glue-to-bone adhesion, dopamine and dopamine/iodinate modified adhesive used for 45° oblique fracture fixation showed good and similar strength at compression (3.05 and 2.79 MPa, respectively). However, significantly higher elasticity of about 250% exhibited adhesive with iodinate enhancer. Moreover, mechanical properties of B2 fractures fixed with both LCP and dopamine/iodinate adhesive were approaching closely to the properties of original bone. Excellent adhesion between the adhesive and the bone fragments was proved by μCT.
The polymer-composite bone adhesive modified with dopamine/iodinate exhibited very good fixation ability of femoral artificial comminuted fractures in an experimental model.
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Modification of thermogelling biodegradable copolymers with functional groups enables further covalent crosslinking of physical micelle‐based hydrogels formed at specific temperature in water. The ...resulting hybrid hydrogel network exhibits an increase in stiffness and degradation stability. In this work, synthesized well‐defined thermoresponsive α,ω‐itaconyl‐poly(d,l‐lactide‐co‐glycolide)‐b‐poly(ethylene glycol)‐b‐poly(d,l‐lactide‐co‐glycolide) (α,ω‐itaconyl‐PLGA‐PEG‐PLGA) macromonomers with a high degree of itaconyl‐substitution providing free double bonds are photocrosslinked in water at both 25 and 37 °C using lithium phenyl‐2,4,6‐trimethylbenzoylphosphinate (LiTPO) acting as water‐soluble non‐toxic photoinitiator. The effect of LiTPO on the thixotropic behavior of macromonomer in water at 25 °C without irradiation is evaluated. With the addition of a low amount of the photoinitiator (0.1 wt%), the degree of copolymer thixotropy increases. However, further increase in the photoinitiator concentration (0.5–3 wt%) leads to a lower degree of thixotropy. The photoinitiator is presumably interfering with the micellar self‐assembly of the copolymer. This trend is also reflected in photocrosslinking efficiency, where the hybrid hydrogel networks with the highest storage moduli are achieved with very low concentrations of the photoinitiator (0.1 wt%) at 25 °C, while this trend is reversed at 37 °C. The hydrolytic stability of hydrogels prepared at 37 °C from 17 wt% macromonomer solution with 1% LiTPO exceeds 22 days.
Additional covalent crosslinking of thermogelling α,ω‐itaconyl‐PLGA‐PEG‐PLGA in water using lithium phenyl‐2,4,6‐trimethylbenzoylphosphinate as a photoinitiator is studied. The photoinitiator influences self‐assembly of the copolymer, that also affects the photocrosslinking efficiency at 25 °C. On the other hand, the photocrosslinking at 37 °C is less affected by the photoinitiator concentration.
Depolarization at high temperatures around the Curie point constitutes an important yet difficultly measurable material property of piezoelectric (PZT) ceramics. The common vibrometric technique (d33 ...meters) is not suitable for the measurement of temperature dependences, and therefore we used the frequency method to perform the desired procedures. The indicator selected to show the depolarization state in the piezoelectric ceramics consisted in the piezoelectric charge coefficient, whose value can be effectively measured via the above-mentioned frequency technique. The accuracy of the method was verified via comparing a d33 meter constructed by the authors (as described in this paper) and also by means of differently sized cylinders of ceramics NCE51, which are designed for longitudinal length modes. Based on the obtained results, we established a measurement methodology to exactly determine the value of the Curie point that corresponded to the phase transition to a cubic crystallographic structure. The experiment also confirmed the applicability of progressively controlled depolarization of PZT ceramics by high temperature in the range of between 350 and 370°C, and it defined the temperature limits at which there occur irreversible changes of the piezoelectric properties of PZT ceramics. In the measured NCE51 material, the limit for irreversible changes was equal to 95% of the Curie temperature (368°C).
Biodegradable thermosensitive triblock copolymers based on poly(ethylene glycol) and poly(lactic‐co‐glycolic acid) (PLGA‐PEG‐PLGA) prepared via ring opening polymerization were modified by itaconic ...anhydride (ITA), which gives copolymer both reactive double bonds and functional carboxylic acid groups essential for the reaction with biological active material. Functionalization conditions comprising ITA purification, temperature, time and presence of solvent were optimized with the respect to amount of end‐capped ITA. Maximum of 76.6 mol. % of bonded ITA were reached via “one pot” reaction in a bulk at 110 °C after 1.5 h. ITA functionalization thermally stabilized the original copolymer by increasing the initial degradation temperature Td from 284 °C to 294 °C and changing the negative glass transition temperature (Tg = ‐1.8 °C) to positive one up to 2.4 °C. The novel functionalized macromonomer can be cross‐linked either chemically or physically in order to produce new functionalized hydrogel network applicable as biomedical material in tissue engineering.
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