Normal and electrically stimulated PC12 cell cultures and the implantation of nerve guidance channels were performed to evaluate newly developed electrically conductive biodegradable polymer composites. Polypyrrole (PPy) doped by butane sulfonic acid showed a significantly higher number of viable cells compared with PPy doped by polystyrenesulfonate after a 6-day culture. The PC12 cells were left to proliferate for 6 days, and the PPy-coated membranes, showing less initial cell adherence, recorded the same proliferation rate as did the noncoated membranes. Direct current electricity at various intensities was applied to the PC12 cell-cultured conductive membranes. After 7 days, the greatest number of neurites appeared on the membranes with a current intensity approximating 1.7-8.4 microA/cm. Nerve guidance channels made of conductive biodegradable composite were implanted into rats to replace 8 mm of sciatic nerve. The implants were harvested after 2 months and analyzed with immunohistochemistry and transmission electron microscopy. The regenerated nerve tissue displayed myelinated axons and Schwann cells that were similar to those in the native nerve. Electrical stimulation applied through the electrically conductive biodegradable polymers therefore enhanced neurite outgrowth in a current-dependent fashion. The conductive polymers also supported sciatic nerve regeneration in rats.
This study evaluated the in vivo biocompatibility and biodegradation behavior of a novel polypyrrole (PPy)/poly(D,L-lactide) (PDLLA) composite and PPy-coated poly(D,L-lactide-co-glycolide) membranes. Test membranes were implanted subcutaneously in rats for 3-120 days. The biocompatibility was assessed by quantifying the alkaline and acid phosphatase secretion, the immunohistochemical staining of the ED-2-positive macrophages, and the histology at the tissue/material interface. The degradation was investigated using scanning electron microscopy. Pure PDLLA and poly(D,L-lactide-co-glycolide) membranes were used as references, whereas expanded polytetrafluoroethylene and a commercial styrene-butadiene rubber were used as controls. The enzyme activity of the PPy-containing specimens was shown to be similar to that of the references. The histological findings were consistent with the enzymatic results, showing a mild-to-moderate acute inflammation followed by a resolution of the inflammatory response with a decrease in inflammatory cells for each biodegradable membrane. The tissue reactions to the PPy, which was either in the form of nanoparticles or surface coating, were comparable to the response to the neighboring biodegradable materials. Elevated ED-2-positive macrophage populations appeared as early as day 3 in the loose connective tissue surrounding the implants. The density of these populations was related to the degree of inflammation. Scanning electron microscopy showed that the degradation of the PPy/PDLLA composite was not affected by the presence of PPy.
The electrical stability of a novel polypyrrole (PPy)/poly(D,L-lactide) (PDLLA) composite was studied in vitro and compared with that of PPy-coated polyester fabrics. Specimens were incubated in Ringer's solution at 37 degrees C for up to 8 weeks with or without the circulation of DC current under a constant 100 mV voltage. In situ current variation with incubation time was recorded. The AC volume electrical conductivity of the specimens before and after incubation in phosphate-buffered saline was recorded using a frequency analyzer. Water absorption and weight loss were monitored metrologically. Changes in the oxidation state of incubated PPy were analyzed with X-ray photoelectron spectroscopy. The morphological changes were observed with scanning electron microscopy, and the glass transition temperature of the PDLLA was investigated using differential scanning calorimetry. The PPy/PDLLA composite in Ringer's solution sustained a relatively stable conductivity up to 8 weeks after an initial period of "conditioning." The PPy-coated fabrics experienced a rapid loss of conductivity when subjected to electrical circulation and regained part of it when disconnected. The volume conductivity of the nonincubated PPy/PDLLA membrane behaved as a typical conductor in the low-frequency range. The mechanisms involved in the various electrical behaviours of the PPy/PDLLA composite and PPy-coated fabrics are discussed. In conclusion, the PPy/PDLLA composite was able to deliver a biologically significant electrical current in a simulated biological solution for up to 8 weeks and therefore may be considered as a first-generation synthetic biodegradable bioconductor.
Biomimetic hydrogel made of poly(ethylene glycol) and soy protein with a water content of 96% has been developed for moist wound dressing applications. In this study, such hybrid hydrogels were investigated by both tensile and unconfined compression measurements in order to understand the relationships between structural parameters of the network, its mechanical properties and protein absorption in vitro. Elastic moduli were found to vary from 1 to 17 kPa depending on the composition, while the Poisson's ratio (approximately 0.18) and deformation at break (approximately 300%) showed no dependence on this parameter. Further calculations yielded the crosslinking concentration, the average molecular weight between crosslinks (M(C)) and the mesh size. The results show that reactions between PEG and protein create polymeric chains comprising molecules of PEG and protein fragments between crosslinks. M(C) is three times higher than that expected for a "theoretical network." On the basis of this data, we propose a model for the 3D network of the hydrogel, which is found to be useful for understanding drug release properties and biomedical potential of the studied material.
The paper describes preparation and biological characterization of the solid hybrid biomaterial that was designed for cell-targeted lipid delivery in healing tissues. The material referred to as 'solid emulsion gel' combines a protein-stabilized lipid emulsion and a hydrogel structure in a single compartment. The potential of the omega-3 (n-3)-fatty acids rich solid emulsion gel for tissue repair applications was investigated at the macro-, micro-, molecular and gene expression levels, using human fibroblasts and endothelial cells and a porcine model of full-thickness wounds. Being non-cytotoxic in vitro and in vivo, the biomaterial was found to affect cell metabolism, modulate expression of certain genes, stimulate early angiogenesis and promote wound repair in vivo. The neovascular response in vivo was correlated with upregulated expression of the genes involved in lipid transport (e.g. adipophilin), anti-apoptosis (e.g. heat shock proteins, haem oxygenase 1) and angiogenesis (vascular endothelial growth factor, placental growth factor). Collectively, the results of this study provide first evidence that the angiogenic response provided by solid emulsion gel-mediated delivery of n-3 fatty acids is an alternative to the topical administration of exogenous growth factors or gene therapy, and can be advantageously used for the stimulation of tissue repair in complex wounds.
The availability of a safe macrofilaricidal drug would help to accelerate onchocerciasis elimination. A trial was conducted in Cameroon to evaluate the effects of a subcutaneous injectable long-acting formulation of ivermectin (LAFI) on the microfilariae (mf) and adult stages of Onchocerca ochengi. Ten zebu cattle naturally infected with the parasite were injected subcutaneously with either 500 mg (group A, N = 4), or 1000 mg long-acting ivermectin (group B, N = 4) or the vehicle (group C, N = 2). Skin samples were collected from each animal before, and 6, 12, and 24 months after treatment to measure microfilarial densities (MFDs). Nodules excised before, and 6 and 12 months after treatment were examined histologically to assess the adult worms’ viability and reproductive status. Blood samples were collected at pre-determined time-points to obtain pharmacokinetic data. Before treatment, the average O. ochengi MFDs were similar in the three groups. Six months after treatment, all animals in groups A and B were free of skin mf, whereas those in group C still showed high MFDs (mean = 324.5 mf/g). Only one ivermectin-treated animal (belonging to group A) had skin mf 12 months after treatment (0.9 mf/g). At 24 months, another animal in group A showed skin mf (10.0 mf/g). The histologic examination of nodules at 6 and 12 months showed that LAFI was not macrofilaricidal but had a strong effect on embryogenesis. The new LAFI regimen might be an additional tool to accelerate the elimination of human onchocerciasis in specific settings.
A series of the solid emulsion gels with the oil volume fraction in the range of 0-50% were synthesized through a polycondensation reaction between activated p-nitrophenyl carbonate poly(ethylene glycol) and protein-stabilized oil-in-water emulsions. The resultant structures were investigated in terms of swelling behavior, composition, morphology, mechanical and skin hydration properties. Solid emulsions gels share the properties of both hydrogel and emulsion. Similar to the classical hydrogel, the SEG swells in water up to equilibrium swelling degree, which decreases as the oil volume fraction increases, and comprises immobilized drops of protein-stabilized oil. The impregnation of the oil phase is found to reduce tensile stiffness of the material, but improves material's extensibility. The mechanical properties of the constructs (Young moduli in the range of 9-15 kPa and the elongation at break of 120-220%) are interpreted according to the "rule of elasticity mixture" that considers the elasticity of the composite material to be a sum of the contributions from individual components, i.e. hydrogel and dispersed oil drops. An idealized model that takes into account the history of the material preparation has been proposed to explain the improved extensibility of the constructs. The results of the mechanical tests, equilibrium swelling, and the skin hydration effect of the solid emulsion gels in vivo are discussed from the perspective of the biomedical applications of the solid emulsion gels, in particular, for the transdermal delivery of hydrophilic and lipophilic drugs.
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