High-yield fabrication of monodisperse multilayer nanofibrous microparticles for advanced oral drug delivery applications.

Recent advances in the use of nano- and microparticles in drug delivery, cell therapy, and tissue engineering have led to increasing attention towards nanostructured microparticulate formulations for maximum benefit from both nano- and micron sized features. Scalable manufacturing of monodisperse nanostructured microparticles with tunable size, shape, content, and release rate remains a big challenge. Current technology, mainly comprises complex multi-step chemical procedures with limited control over these aspects. Here, we demonstrate a novel technique for high-yield fabrication of monodisperse monolayer and multilayer nanofibrous microparticles (MoNami and MuNaMi respectively). The fabrication procedure includes sequential electrospinning followed by micro-cutting at room temperature and transfer of particles for collection. The big advantage of the introduced technique is the potential to apply several polymer-drug combinations forming multilayer microparticles enjoying extracellular matrix (ECM)-mimicking architecture with tunable release profile. We demonstrate the fabrication and study the factors affecting the final three-dimensional structure. A model drug is encapsulated into a three-layer sheet (PLGA-pullulan-PLGA), and we demonstrate how the release profile changes from burst to sustain by simply cutting particles out of the electrospun sheet. We believe our fabrication method offers a unique and facile platform for realizing advanced microparticles for oral drug delivery applications.

Advancing autologous urothelial micrografting and composite tubular grafts for future single-staged urogenital reconstructions.

Urogenital reconstructive surgery can be impeded by lack of tissue. Further developments within the discipline of tissue engineering may be part of a solution to improve clinical outcomes. In this study, we aimed to design an accessible and easily assembled tubular graft with autologous tissue, which could be constructed and implanted as a single-staged surgical procedure within the premises of an ordinary operating room. The ultimate goals would be to optimize current treatment-options for long-term urinary diversion. Therefore, we evaluated the optimal composition of a collagen-based scaffold with urothelial micrografts in vitro, and followingly implanted the construct in vivo as a bladder conduit. The scaffold was evaluated in relation to cell regeneration, permeability, and biomechanical properties. After establishing an optimized scaffold in vitro, consisting of high-density collagen with submerged autologous micrografts and reinforced with a mesh and stent, the construct was successfully implanted in an in vivo minipig model. The construct assemblance and surgical implantation proved feasible within the timeframe of a routine surgical intervention, and the animal quickly recovered postoperatively. Three weeks post-implantation, the conduit demonstrated good host-integration with a multilayered luminal urothelium. Our findings have encouraged us to support its use in more extensive preclinical large-animal studies.

Gelatin from Saithe (Pollachius virens) Skin: Biochemical Characterization and Oxidative Stability in O/W Emulsions.

This study performed the extraction of gelatin from saithe (Pollachius virens) skin and compared it to commercial marine gelatin. As a first stage, we investigated the physicochemical and biochemical properties of the gelatin. SDS-PAGE analysis revealed the presence of α-chains, ß-chains, and other high-molecular-weight aggregates. DSC thermograms showed typical gelatin behavior, while the FTIR spectra were mainly situated in the amide band region (amide A, amide B, amide I, amide II, and amide III). In the second stage, we produced O/W emulsions and analyzed their physical and oxidative stability over 9 days. Oil droplets stabilized with the gelatins obtained from saithe fish skin had a size of ~500 nm and a ζ-potential ~+25 mV, which is comparable to oil droplets stabilized with commercial gelatin products. Moreover, the oxidative stability of the emulsions stabilized with gelatin from saithe fish skin showed promising results in terms of preventing the formation of some volatile compounds towards the end of the storage period compared to when using the commercial gelatins. This study indicates the potential application of fish skin gelatin in the fields of food and cosmetics, as well as suggesting that further investigations of their techno-functional properties.

Delivery of streptomycin to the rat colon by use of electrospun nanofibers.

Drug-loaded electrospun nanofibers are potential drug carrier systems that may optimize disease treatment while reducing the impact on commensal microbes. The feasibility of streptomycin-loaded pullulan nanofibers fabricated from a green electrospinning procedure using water as the solvent was assessed. We conducted a rat study including a group treated with streptomycin-loaded nanofibers (STR-F, n = 5), a group treated with similar concentrations of streptomycin in the drinking water (STR-W, n = 5), and a non-treated control group (CTR, n = 5). Streptomycin was successfully loaded into nanofibers and delivered by this vehicle, which minimized the quantity of the drug released in the ileal compartment of the gut. Ingested streptomycin-resistant E. coli colonized of up to 106 CFU/g feces, revealing a selective effect of streptomycin even when given in the low amounts allowed by the nanofiber-based delivery. 16S amplicon sequencing of the indigenous microbiota revealed differential effects in the three groups. An increase of Peptostreptococcaceae in the cecum of STR-F animals may indicate that the fermentation of nanofibers directly or indirectly promoted growth of bacteria within this family. Our results elucidate relevant properties of electrospun nanofibers as a novel vehicle for delivery of antimicrobials to the large intestine.

Electrospun composite nanofibers as novel high-performance and visible-light photocatalysts for removal of environmental pollutants: A review.

Environmental pollution caused by industries and human manipulations is coming a serious global challenge. On the other hand, the world is facing an energy crisis caused by population growth. Designing solar-driven photocatalysts which are inspired by the photosynthesis of plant leaves is a fantastic solution to use solar energy as green, available, and unlimited energy containing ∼50% visible light for the removal of environmental pollutants. The polymeric and non-polymeric-based electrospun composite nanofibers (NFs) are as innovative photocatalytic candidates which increase photocatalytic activity and transition from UV light to visible light and overcome the aggregation, photocorrosion, toxicity, and hard recycling and separation of the nanosized powder form of photocatalysts. The composite NFs are fabricated easily by either embedding the photocatalytic agents into the NFs during electrospinning or via their decorating on the surface of NFs post-electrospinning. Polyacrylonitrile-based, tungsten trioxide-based, zinc oxide-based, and titanium dioxide-based composite NFs were revealed as the most reported composite NFs. All the lately investigated electrospun composite NFs indicated long-term stability, high photocatalytic efficiency (∼> 80%) within a short time of light radiation (10-430 min), and high stability after several cycles of use. They were applied in various applications including degradation of dyes/antibiotics, water splitting, wastewater treatment, antibacterial usage, etc. The photogenerated species especially holes, O2∙-, and .OH were mostly responsible for the photocatalytic mechanism and pathway. The electrospun composite NFs have the potential to use in large-scale productions in condition that their thickness and recycling conditions are optimized, and their toxicity and detaching are resolved.

Physical and Oxidative Stability of Low-Fat Fish Oil-in-Water Emulsions Stabilized with Black Soldier Fly (Hermetia illucens) Larvae Protein Concentrate.

The physical and oxidative stability of fish oil-in-water (O/W) emulsions were investigated using black soldier fly larvae (BSFL) (Hermetia illucens) protein concentrate as an emulsifier. To improve the protein extraction and the techno-functionality, defatted BSFL powder was treated with ohmic heating (BSFL-OH) and a combination of ohmic heating and ultrasound (BSFL-UOH). Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) were performed in order to characterize the secondary structure and thermal stability of all protein concentrate samples. The interfacial properties were evaluated by the pendant drop technique. The lowest interfacial tension (12.95 mN/m) after 30 min was observed for BSFL-OH. Dynamic light scattering, ζ-potential and turbiscan stability index (TSI) were used to evaluate the physical stability of emulsions. BSFL-OH showed the smallest droplet size (0.68 µm) and the best emulsion stability (TSI = 8.89). The formation of primary and secondary volatile oxidation products and consumption of tocopherols were evaluated for all emulsions, revealing that OH and ultrasound treatment did not improve oxidative stability compared to the emulsion with untreated BSFL. The results revealed the promising application of BSFL proteins as emulsifiers and the ability of ohmic heating to improve the emulsifying properties of BSFL proteins.

Physico-chemical and colloidal properties of protein extracted from black soldier fly (Hermetia illucens) larvae.

The black soldier fly larvae (BSFL), Hermetia illucens (Linnaeus), has been largely utilized for animal feed. Due to its interesting composition, BSFL has great potential to be further implemented in the human diet. Herein we compared the flour and protein extract composition based on their moisture, ash, amino acids, mineral, and protein content. To have wide knowledge on protein profile and behavior, SDS-page electrophoresis, Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) were used to give information about protein structure and thermal stability, respectively. The flour and protein extract contained respectively 37.3% and 61.1% of protein. DSC graph reported a glass transition temperature around 30 °C, recognizable by a shift in the curve, and an endothermic peak for solid melting at around 200 °C. FTIR analysis showed the main amide bands (A, B, I, II, III) for the flour and protein extract. The foam properties of BSFL protein extract were explored under different temperatures treatment, and the best foam stability was reached at 85 °C with 15 min of treatment. The data highlight the promising techno-functional properties of BSFL protein extract, and that the nutritional composition might be suitable for further use of BSFL as food fortification system.

The determinant role of fabrication technique in final characteristics of scaffolds for tissue engineering applications: A focus on silk fibroin-based scaffolds.

3D scaffolds are in the center of attention for tissue engineering applications. Whilst many studies have focused on the biological properties of scaffolds, less attention has been paid to meeting the biomechanics of the target tissues. In this work, we show how using the same original biomaterial, but different fabrication techniques can lead to a broad range of structural, mechanical, and biological characteristics. Starting with silk fibroin filament as our base biomaterial, we employed electrospinning, film casting, and weft knitting as different scaffold fabrication techniques. Among these three, the weft knit scaffold showed outstanding cell-scaffold interaction including full 3D cell attachment, complete cell coverage around individual filaments, and in-depth cell infiltration. Post-fabrication degumming of silk filament yarns resulted in more bulky and less open pores for the silk fibroin knit scaffold. The decreased pore size after degumming of knit scaffold alleviated the need to in-advance pore filling (a requisite for increasing cell adhesion in a typical knit scaffold having big pores). From a mechanical viewpoint, the weft knit scaffold shows the highest mechanical strength alongside with far better extensibility. Interestingly, the silk filament weft knit scaffold (in the course direction) was 100 and 1000 times more compliant than silk fibroin film and electrospun web, respectively. The observed effect of material type and fabrication technique highlights the suitability of silk fibroin weft-knit scaffolds for the regeneration of load-bearing soft tissues such as urine bladder.

Polymeric carriers for enhanced delivery of probiotics.

Probiotics are live microorganisms (usually bacteria), which are defined by their ability to confer health benefits to the host, if administered adequately. Probiotics are not only used as health supplements but have also been applied in various attempts to prevent and treat gastrointestinal (GI) and non-gastrointestinal diseases such as diarrhea, colon cancer, obesity, diabetes, and inflammation. One of the challenges in the use of probiotics is putative loss of viability by the time of administration. It can be due to procedures that the probiotic products go through during fabrication, storage, or administration. Biocompatible and biodegradable polymers with specific moieties or pH/enzyme sensitivity have shown great potential as carriers of the bacteria for 1) better viability, 2) longer storage times, 3) preservation from the aggressive environment in the stomach and 4) topographically targeted delivery of probiotics. In this review, we focus on polymeric carriers and the procedures applied for encapsulation of the probiotics into them. At the end, some novel methods for specific probiotic delivery, possibilities to improve the targeted delivery of probiotics and some challenges are discussed.

Single particles as resonators for thermomechanical analysis.

Thermal methods are indispensable for the characterization of most materials. However, the existing methods require bulk amounts for analysis and give an averaged response of a material. This can be especially challenging in a biomedical setting, where only very limited amounts of material are initially available. Nano- and microelectromechanical systems (NEMS/MEMS) offer the possibility of conducting thermal analysis on small amounts of materials in the nano-microgram range, but cleanroom fabricated resonators are required. Here, we report the use of single drug and collagen particles as micro mechanical resonators, thereby eliminating the need for cleanroom fabrication. Furthermore, the proposed method reveals additional thermal transitions that are undetected by standard thermal methods and provide the possibility of understanding fundamental changes in the mechanical properties of the materials during thermal cycling. This method is applicable to a variety of different materials and opens the door to fundamental mechanistic insights.

Thread-Like Radical-Polymerization via Autonomously Propelled (TRAP) Bots.

Micromotor-mediated synthesis of thread-like hydrogel microstructures in an aqueous environment is presented. The study utilizes a catalytic micromotor assembly (owing to the presence of a Pt layer), with an on-board chemical reservoir (i.e., polymerization mixture), toward thread-like radical-polymerization via autonomously propelled bots (i.e., TRAP bots). Synergistic coupling of catalytically active Pt layer, together with radical initiators (H2 O2 and FeCl3 (III)), and PEGDA monomers preloaded into the TRAP bot, results in the polymerization of monomeric units into elongated thread-like hydrogel polymers coupled with self-propulsion. Interestingly, polymer generation via TRAP bots can also be triggered in the absence of hydrogen peroxide for cellular/biomedical application. The resulting polymeric hydrogel microstructures are able to entrap living cells (NIH 3T3 fibroblast cells), and are easily separable via a centrifugation or magnetic separation (owing to the presence of a Ni layer). The cellular biocompatibility of TRAP bots is established via a LIVE/DEAD assay and MTS cell proliferation assay (7 days observation). This is the first study demonstrating real-time in situ hydrogel polymerization via an artificial microswimmer, capable of enmeshing biotic/abiotic microobjects in its reaction environment, and lays a strong foundation for advanced applications in cell/tissue engineering, drug delivery, and cleaner technologies.

Naturally-derived electrospun wound dressings for target delivery of bio-active agents.

Electrospun nanofibers are known as the advanced means for wound dressing. They have represented remarkable potency to encapsulate and deliver biomolecules promoting the wound healing process. Compared to synthetic polymers, naturally derived polymers (NDP) are more qualified candidates for fabrication of biomedical electrospun scaffolds. Not only nanofibers of NDP illustrate higher biocompatibility and biodegradability rates, but also they mimic the native extracellular matrix more closely, which leads to the wound closure acceleration by enhancing tissue regeneration. Aside, incorporation of bioactive molecules and therapeutic agents into the nanofibers can generate innovative bioactive wound dressings with significantly improved healing potentials. This paper starts with a brief discussion on the steps and factors influencing the wound healing process. Then, the recent applications of electrospun nanofibers as wound dressing with healing accelerating properties are reviewed. Further, the various healing agents and alternative strategies for modification and functionalization of bioactive naturally-derived electrospun nanofibers are discussed.

In vitro permeability enhancement of curcumin across Caco-2 cells monolayers using electrospun xanthan-chitosan nanofibers.

Xanthan-Chitosan (X-Ch) polysaccharides nanofibers were prepared using electrospinning processing as an encapsulation and delivery system of curcumin (Cu). The X-Ch-Cu nanofibers remained stable in aqueous HBSS medium at pH 6.5 and pH 7.4, mainly due to the ability of oppositely charged xanthan-chitosan polyelectrolytes to form ionically associated electrospun nanofibers. The xanthan-chitosan-curcumin nanofibers were incubated with Caco-2 cells, and the cell viability, transepithelial transport and permeability properties across cell monolayers were investigated. After 24 h of incubation, the exposure of Caco-2 cell monolayers to X-Ch-Cu nanofibers resulted in a cell viability of ∼80%. A 3.4-fold increase of curcumin permeability was observed when the polyphenol was loaded into X-Ch nanofibers, compared to the free curcumin. This increased in vitro transepithelial permeation of curcumin without compromising cellular viability was induced by interactions upon contact between the nanofibers and the Caco-2 cells, leading to the opening of the tight junctions. The results obtained revealed that X-Ch nanofibers can be used for oral delivery applications of poorly water-soluble compounds at the gastrointestinal tract.

Synthesis and therapeutic potential of silver nanomaterials derived from plant extracts.

Silver nanoparticles (AgNPs) have attracted a great deal of attention in the recent years. It is mostly due to their availability, chemical stability, catalytic activity, conductivity, biocompatibility, antimicrobial activity and intrinsic therapeutic properties. There are three major approaches for AgNPs synthesis; i.e., chemical, physical, and biological methods. Today, many of chemical and physical methods have become less popular due to using hazardous chemicals or their high costs, respectively. The biological method has introduced an appropriate substitute synthesis strategy for the traditional physical and chemical approaches. The utilization of the plant extracts as reducing, stabilizing and coating agent of AgNPs is an interesting eco-friendly approach leading to high efficiency. The antimicrobial and anticancer synergistic effects among the AgNPs and phytochemicals will enhance their therapeutic potentials. Surprisingly, although many studies have demonstrated the significant enhancement in cytotoxic activities of plant-mediated AgNPs toward cancerous cells, these nanoparticles have been found nontoxic to normal human cells in their therapeutic concentrations. This review provides a comprehensive insight into the mechanism of plant-mediated AgNPs synthesis, their antimicrobial and cytotoxic activities as well as their applications.

Bladder biomechanics and the use of scaffolds for regenerative medicine in the urinary bladder.

The urinary bladder is a complex organ with the primary functions of storing urine under low and stable pressure and micturition. Many clinical conditions can cause poor bladder compliance, reduced capacity, and incontinence, requiring bladder augmentation or use of regenerative techniques and scaffolds. To replicate an organ that is under frequent mechanical loading and unloading, special attention towards fulfilling its biomechanical requirements is necessary. Several biological and synthetic scaffolds are available, with various characteristics that qualify them for use in bladder regeneration in vitro and in vivo, including in the treatment of clinical conditions. The biomechanical properties of the native bladder can be investigated using a range of mechanical tests for standardized assessments, as well as mathematical and computational bladder biomechanics. Despite a large body of research into tissue engineering of the bladder wall, some features of the native bladder and the scaffolds used to mimic it need further elucidation. Collection of comparable reference data from different animal models would be a helpful tool for researchers and will enable comparison of different scaffolds in order to optimize characteristics before entering preclinical and clinical trials.

Bladder wall biomechanics: A comprehensive study on fresh porcine urinary bladder.

Regenerative medicine for reconstructive urogenital surgery has been widely studied during the last two decades. One of the key factors affecting the quality of bladder regeneration is the mechanical properties of the bladder scaffold. Insight into the biomechanics of this organ is expected to assist researchers with functional regeneration of the bladder wall. Due to extensive similarities between human bladder and porcine bladder, and with regard to lack of comprehensive biomechanical data from the porcine bladder wall (BW), our main goal here was to provide a thorough evaluation on viscoelastic properties of fresh porcine urinary BW. Three testing modes including Uniaxial tensile, ball-burst (BB) and Dynamic Mechanical Analysis (DMA) were applied in parallel. Uniaxial tests were applied to study how different circumferential and longitudinal cut-outs of lateral region of BW behave under load. DMA was used to measure the viscoelastic properties of the bladder tissue (storage and loss modulus) in a frequency range of 0.1-3Hz. BB was selected as a different technique, replicating normal physiological conditions where the BW is studied in whole. According to uniaxial tests, the anisotropic behavior of bladder is evident at strain loads higher than 200%. According to DMA, storage modulus is consistently higher than loss modulus in both directions, revealing the elasticity of the BW. The stress-strain curves of both uniaxial and BB tests showed similar trends. However, the ultimate stress measured from BB was found to be around 5 times of the relevant stress from uniaxial loading. The ultimate strain in BB (389.9 ± 59.8) was interestingly an approximate average of rupture strains in longitudinal (358 ± 21) and circumferential (435 ± 69) directions. Considering that each testing mode applied here reveals distinct information, outcomes from the combination of the three can be considered as a helpful data-base to refer to for researchers aiming to regenerate the bladder.