Wound healing is a process regulated by a complex interaction of multiple growth factors including fibroblast growth factor 2 (FGF2). Although FGF2 appears in several tissue engineered studies, its applications are limited due to its low stability both in vitro and in vivo. Here, this shortcoming is overcome by a unique nine-point mutant of the low molecular weight isoform FGF2 retaining full biological activity even after twenty days at 37 °C. Crosslinked freeze-dried 3D porous collagen/chitosan scaffolds enriched with this hyper stable recombinant human protein named FGF2-STAB® were tested for in vitro biocompatibility and cytotoxicity using murine 3T3-A31 fibroblasts, for angiogenic potential using an ex ovo chick chorioallantoic membrane assay and for wound healing in vivo with 3-month old white New Zealand rabbits. Metabolic activity assays indicated the positive effect of FGF2-STAB® already at very low concentrations (0.01 µg/mL). The angiogenic properties examined ex ovo showed enhanced vascularization of the tested scaffolds. Histological evaluation and gene expression analysis by RT-qPCR proved newly formed granulation tissue at the place of a previous skin defect without significant inflammation infiltration in vivo. This work highlights the safety and biocompatibility of newly developed crosslinked collagen/chitosan scaffolds involving FGF2-STAB® protein. Moreover, these sponges could be used as scaffolds for growing cells for dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration.
Patients with critical thermal trauma belong to one of the most high-risk groups for development of infectious complications. Fungal infections are not among frequent complications during therapy of patients with thermal trauma, yet their incidence dramatically aggravates the prognosis for patients with this disorder. In the case report, we present the case of a young man with a critical burn, where Westerdykella dispersa was isolated. Identification of the pathogen was provided with a combination of cultivation and molecular biological confirmation. In this case, the distinction between infection and colonization was very complicated. Histopathological examination for definitive diagnosis of infection was not performed because the material from unburned soft tissue sampling could further compromise the function of the hand. Repeated cultivation and molecular identification W. dispersa before and after the necrectomy is indicative of infection rather than colonization. It is the second documented case of positive cultivation with this pathogen in humans and the first such case in a non-neutropenic host.
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.
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.
Toxic epidermal necrolysis (TEN) is a rare disease, which predominantly manifests as damage to the skin and mucosa. Antibiotics count among the most common triggers of this hypersensitive reaction. Patients with TEN are highly susceptible to infectious complications due to the loss of protective barriers and immunosuppressant therapy. The aim of this study was to investigate the potential relationship between antibiotics used before the development of TEN and early and late-onset infectious complications in TEN patients. In this European multicentric retrospective study (Central European Lyell syndrome: therapeutic evaluation (CELESTE)), records showed that 18 patients with TEN used antibiotics (mostly aminopenicillins) before the disease development (group 1), while in 21 patients, TEN was triggered by another factor (group 2). The incidence of late-onset infectious complications (5 or more days after the transfer to the hospital) caused by Gram-positive bacteria (especially by Enterococcus faecalis/faecium) was significantly higher in group 1 than in group 2 (82.4% vs. 35.0%, p = 0.007/pcorr = 0.014) while no statistically significant difference was observed between groups of patients with infection caused by Gram-negative bacteria, yeasts, and filamentous fungi (p > 0.05). Patients with post-antibiotic development of TEN are critically predisposed to late-onset infectious complications caused by Gram-positive bacteria, which may result from the dissemination of these bacteria from the primary focus.
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