Design of Volumetric Nanolayers via Rapid Proteolysis of Silk Fibroin for Tissue Engineering.

Various methods have been studied to make a regenerated silk fibroin solution. However, most of them take too much time and effort to liquefy. Here, we report that a regenerated silk fibroin solution could be prepared within seconds through acid proteolysis for the first time. The solubilized fibroin could be applied to advanced tissue engineering. Our method shortened the production time to one day (more than 10 times) compared to the general fibroin solution preparation method. It was confirmed that the initial protein affinity nearly doubled from 0.028 to 0.076 µg·mm-2 in FF(ac) compared to FF(aq). A fibroin nanofiber layer having a volumetric hierarchical structure was prepared by electrospinning an acid-proteolyzed fibroin solution, followed by gas foaming. In vitro results of cell adhesion and proliferation capacity of the gas-foamed scaffold were not significantly different compared to the two-dimensional (2D) fibroin nanofiber membrane, overcoming the limitations of volumetric nanofiber scaffolds. We are confident that our research will greatly contribute to the development of regenerative engineering using other proteins.

Near-Infrared Responsive Synergistic Chemo-Phototherapy from Surface-Functionalized Poly(ε-caprolactone)-Poly(d,l-lactic-co-glycolic acid) Composite Nanofibers for Postsurgical Cancer Treatment.

The combination of hyperthermia and chemotherapy has attracted significant attention in local cancer treatment following surgical resection. Pyrrole is a potent photothermal agent that can induce a temperature rise at different concentrations in the surrounding medium by absorbing near-infrared radiation (NIR). In this study, poly(ε-caprolactone) (PCL) and poly (d,l-lactic-co-glycolic acid) (PLGA) were used to make nanofibers using the electrospinning process. Then, pyrrole in different concentrations of (0.2, 0.4, and 0.6) M was attached to the surface of PCL-PLGA fiber mats by in situ polymerization, which was confirmed by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction (XRD) analysis. A concentration-dependent local temperature rise was observed using a FLIR camera under near-infrared (NIR) laser irradiation. For the hyperthermia effect, pyrrole concentration (0.06 M) was used for in vitro drug release studies and cell viability assays because under NIR irradiation (2 W/cm2, 3 min), it increased the local temperature to around 45 °C. In vitro drug release studies confirmed that NIR irradiation increased the diffusion rate of doxorubicin (DOX) by increasing the environmental temperature above the glass transition temperature of PLGA. In vitro cytotoxicity experiments further confirmed that PCL-PLGA-DOX/PPy fiber mats showed an enhanced inhibitory effect against CT26 and MCF7 cells by the combination of hyperthermia and chemotherapy.

Silver nanoparticles decorated reduced graphene oxide: Eco-friendly synthesis, characterization, biological activities and embryo toxicity studies.

This study was aimed on the eco-friendly synthesis of silver nanoparticles (AgNPs), reduced graphene oxide (rGO) and AgNPs decorated rGO (rGO/AgNPs) nanocomposite and appraisal of their bioactivities and toxicity. As-prepared nanomaterials were established through high resolution X-ray diffraction (HR-XRD), high resolution transmission electron microscopy (HR-TEM), X-ray photoelectron spectroscopy (XPS), Raman spectroscopy, UV-Vis. spectroscopy and Fourier transform infrared spectroscopy (FT-IR). In this study, leaves extract, graphene oxide (GO) and rGO did not show antibacterial and anticancer activities; no significant embryo toxicity was recorded. On the other hand, AgNPs displayed good antibacterial and anticancer activities; however, higher toxic effects were observed even at the lowest test concentration (0.7 µg/ml). In case of rGO/AgNPs nanocomposite, significant antibacterial activity together with low cytotoxicity was noticed. Interestingly, the embryo toxicity of AgNPs was significantly reduced by rGO, implying the biocompatible nature of as-synthesized nanocomposite. Taken together, these results clearly suggest that rGO/AgNPs nano hybrid composite could be developed as the promising biomaterial for future biomedical applications.

A Study on the Effectiveness of Spatial Filters on Thermal Image Pre-Processing and Correlation Technique for Quantifying Defect Size.

Thermal imaging plays a vital role in structural health monitoring of various materials and provides insight into the defect present due to aging, deterioration, and fault during construction. This study investigated the effectiveness of spatial filters during pre-processing of thermal images and a correlation technique in post-processing, as well as exploited its application in non-destructive testing and evaluation of defects in steel structures. Two linear filters (i.e., Gaussian and Window Averaging) and a non-linear filter (i.e., Median) were implemented during pre-processing of a pulsed thermography image sequence. The effectiveness of implemented filters was then assessed using signal to noise ratio as a quality metric. The result of pre-processing revealed that each implemented filter is capable of reducing impulse noise and producing high-quality images; additionally, when comparing the signal to noise ratio, the Gaussian filter dominated both Window Averaging and Median filters. Defect size was determined using a correlation technique on a sequence of pulsed thermography images that had been pre-processed with a Gaussian filter. Finally, it is concluded that the correlation technique could be applied to the fast measurement of defect size, even though the accuracy may depend on the detection limit of thermography and defect size to depth ratio.

Simple Colorimetric and Fluorescence Chemosensing Probe for Selective Detection of Sn2+ Ions in an Aqueous Solution: Evaluation of the Novel Sensing Mechanism and Its Bioimaging Applications.

An easily accessible colorimetric and fluorescence probe 4-((3-chloro-1,4-dioxo-1,4-dihydronaphthalen-2-yl)amino)benzenesulfonamide (4CBS) was successfully developed for the selective and sensitive detection of Sn2+ in an aqueous solution. The sensing mechanism involves reduction of -C═O into -C-OH groups in 4CBS upon the addition of Sn2+, which initiates the fluorescence turn-on mode. A better linear relationship was achieved between fluorescence intensity and Sn2+ concentration in the range of 0-62.5 µM, with a detection limit (LOD) of 0.115 µM. The binding mechanism of 4CBS for Sn2+ was confirmed by Fourier transform infrared analysis, NMR titrations, and mass (electrospray ionization) spectral analysis. Likewise, the proposed sensing mechanism was supported by quantum chemical calculations. Moreover, bioimaging studies demonstrated that the chemosensing probe 4CBS is an effective fluorescent marker for the detection of Sn2+ in living cells and zebrafish. Significantly, 4CBS was able to discriminate between Sn2+ in human cancer cells and Sn2+ in normal live cells.

Merging 3D printing with electrospun biodegradable small-caliber vascular grafts immobilized with VEGF.

The major challenge of commercially available vascular substitutes comes from their limitations in terms of hydrophobic surface, which is hostile to cell growth. To date, tissue-engineered and synthetic grafts have not translated well to clinical trials when looking at small diameters. We conceptualized a cell-free structurally reinforced biodegradable vascular graft recapitulating the anisotropic feature of a native blood vessel. The nanofibrous scaffold is designed in such a way that it will gradually degrade systematically to yield a neo-vessel, facilitated by an immobilized bioactive molecule-vascular endothelial growth factor (VEGF). The nano-topographic cue of the device is capable of direct host cell infiltration. We evaluated the burst pressure, histology, hemocompatibility, compression test, and mechanical analysis of the new graft. The graft implanted into the carotid artery of a porcine model demonstrated a good patency rate as early as two week post-implantation. This graft reinforced design approach when employed in vascular tissue engineering might strongly influencing regenerative medicine.

Structural Packaging Technique Using Biocompatible Nanofiber with Essential Oil to Prolong the Shelf-Life of Fruit.

We have studied the technique for efficient prolong the shelf-life of fruit. To prolong the shelf-life of fruit, one of the most important factor is the concentration of oxygen (O2) and carbon dioxide (CO2) produced by the fruits inside the packaging. Therefore, to create the right environment, it recommended creating pores which O2 and CO2 can be exchanged. We utilize electrospinning technique for easy electrohydrodynamic fabrication of nanofibers by electric force to fabricate nano-sized fibers with high porosity. Additionally, use of essential oil is very appropriate to reduce postharvest disease cause of their antibacterial and like-pesticide properties. In this study, we fabricated different structural packaging types using nanofibers and essential oil to compare the efficacy. After that, we observed the percentage of corruption is significantly reduced by using peach as a kind of fruits. These packaging techniques will overcome the limitation of fruits life with postharvest disease not only field of fruit packaging techniques.

Fabrication of a Micro/Nano-Net Membrane Using Cellulose Nanocrystals Derived from Seaweed.

We have successfully extracted cellulose nanocrystals (CNCs) from seaweed by removing unwanted materials via our modified process. The prepared CNCs were mixed with two biocompatible polymers (polyethylene oxide (PEO)/Eudragit S100). We used the most popular electrospinning method to fabricate a micro/nano-net membrane. The formation of nano-webs between fundamental micro/nanofibers was observed via SEM and TEM, according to the mixing ratio of the solution (PEO:Eudragit: 1:1, 2:1, 3:1 wt:wt%) with 0, 5, 10% CNCs per polymer weight. We found the optimal condition to fabricate nano-net in the membrane and expect it to be applicable for wound healing, tissue engineering, and various filter applications.

QCN-Based Analysis for Predicting the Quality of Resulting Electrospun Nanofiber: Effect of Real-Time Transient Rheological Properties of Precursor Solution on Electrospinning.

There is a large margin between the mechanical properties and morphology of electrospun fibers required in each area. The produced fibers show a large difference depending on the external environment such as temperature, humidity, and season even in optimum concentration and same electrospinning set-up. The properties of polymer solution among the parameters are the largest determinant of the mechanical strength and diameter of electrospun fibers. Herein, the accurate predicting system in advance to electrospinning is required. In this study, we conduct a comparative study on the viscosity (measured by Brookfield rheometer) and the transient mass change and evaporation speed by our lab-made QCN in order to establish a predicting system for the quality of fiber. It was possible to measure the change of mass of the polymer solution in real-time using the lab-made QCN as well as calculating the volatility, the evaporation time of the polymer solution. The volatility of the polymer solution has a significant impact on the quality of the electrospun fiber including the diameter, uniformity, and surface topography. We compare the mass changes, viscosimetric viscosities, and the quality of corresponding fiber, and reveal the potential of QCN as a tool predicting pre-electrospinning fibers.

Investigation of Composite Nano Air Filter for Improving Antimicrobial Activity and Reducing VOCs Using a High Speed Upward Electrospinning System.

A new nano air filter for fine dust filtration with antibacterial and volatile organic compounds (VOCs) adsorption properties was fabricated using a bottom-up, high-speed electrospinning system. To optimize production, polyurethane fibers were electrospun at various voltages on polypropylene nonwoven fabrics, and results show that fiber diameter decreased as voltage increased. Silver nanoparticles (AgNPs) and Activated Carbon (AC) were used as antimicrobials and VOC-reducing agents. FTIR, SEM, and EDS were performed to analyze the resulting filter fabricated by electrospinning. FTIR and EDS results show that the AgNPs and activated carbon added to the PU fibers were successfully integrated into the PP nonwoven fabric.

Preliminary Study for Measurement of Shear Stress and Hemocompatibility Using Commercialized Lab on a Chip.

We have investigated the effect of flow rate on shear stress and in turn thrombus formation on a lab-on-a-chip with a microchannel that is suitable for cell culture and growth. Using a combination of Arduino UNO, Arduino Motor Shield, and a SERVO stepper motor, we created a pump system that closely mimics the in vivo conditions of the human body. With this system, we achieved continuous flow of blood and observed attached platelets at the bottom of the collagen coated microslide, confirming that with shear stress, thrombus formation increases.

Nano-Nets Covered Composite Nanofibers with Enhanced Biocompatibility and Mechanical Properties for Bone Tissue Engineering.

Enhancing the biocompatibility profiles including cell attachment, growth, and viability and mechanical properties of designed synthetic scaffolds have an essential role in tissue engineering applications. Polymer blending is one of the most effective methods for providing new anticipated biomaterials for tissue scaffolds. Here, the blend solution of the different mass weight ratio of polycaprolactone (PCL) to human serum albumin (HSA) was subjected to fabricate nanocomposite spider-web-like membranes using electrospinning process. The physicochemical aspects of fabricated membranes had been characterized by a different state of techniques like that of scanning electron microscopy (FE-SEM), Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), contact angle meter and universal testing machine. FE-SEM images revealed that all PCL/HSA mats were composed of interlinked nano-nets along with conventional electrospun fibers while nano-nets were not found for pristine PCL mat. Moreover, composite membranes exhibited improved water absorbability, enhanced biodegradation compared to pristine PCL membrane and had much better mechanical properties (tensile strength increased by up to 3-fold, Young's modulus by 2-fold). The cell attachment and proliferation tests were carried by culturing Mc3T3-E1 (pre-osteoblasts) with the designated nanofibrous membranes. The hybrid nanofibers exhibited extraordinary support for the adhesion and proliferation of cells when compared to the pristine PCL membrane. These results indicate that the nano-nets supported PCL/HSA scaffolds can be promising for tissue engineering applications.

Development of Nanofiber Reinforced Double Layered Cabin Air Filter Using Novel Upward Mass Production Electrospinning Set Up.

Recently, numerous researchers are interested in the development of new air filter because of air pollution caused by rapid industrialization and urbanization. The major concerns in developing air filters are: pressure drop and filtration efficiency which are considered significant. As the pressure drop increases, the energy consumption becomes high. In this study, we developed a novel air filter (polyurethane fiber mat) for nano size filtration using a mass production electrospinning, which is expected to enhance filtration efficiency and pressure drop effects. To determine the optimal electrospinning conditions for filter efficiency, various concentrations (8, 10, 12 wt/wt%) of thermoplastic polyurethane were prepared and employed. Scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FT-IR) were used for fiber characterization, and finally, efficiency test was conducted to evaluate the filter performance of developed nanofiber-based air filter. From this study, it could be concluded that optimization by adjusting the polymer concentration and electrospinning operating condition was the best efficient alternative method to fabricate nano-fibrous air filter system with improved filtration performance.

Harnessing Nanotopography of Electrospun Nanofibrous Nerve Guide Conduits (NGCs) for Neural Tissue Engineering.

The anatomical recovery of nerve defects with their neurological functions after an injury caused by diseases or accidents is an important clinical issue. The most efficient surgical technique so far to the nerve defects, which are unrepairable by direct end-to-end suture, can be autograft transplantation. The autograft transplantation, however, has disadvantages including multiple rounds of surgery, a shortage of nerve donor, and function loss at the donor site. Tissue-engineered nerve guide conduits (TENGCs) have emerged as a potential alternative to autologous nerve grafts for nerve regeneration and functional recovery. Various TENGCs researches are being carried out to improve characteristics and enhance functionality such as material selection, biomimetic, topography, and enhancement by the biomolecules additions. Among them, the customizable surface nanotopography of aligned fibrous TENGCs foster neural repair by providing a cell-friendly environment, permissiveness, guidance cues, and directional growth of the cells. Fibrous nerve guide conduits (NGCs) made of longitudinally ordered fibers is a promising candidate for nerve tissue engineering.

Optimization of Electropolishing on NiTi Alloy Stents and Its Influence on Corrosion Behavior.

Nitinol or NiTi alloys are well-known as an attractive biomedical material due to their unique properties such as the shape memory effect, super-elasticity and biocompatibility. These characteristics enable them to be best candidates for implant materials such as stent. One of the major factors that strongly affect the performance of nitinol stent is its unique surface properties. In this study, the influence of electropolishing on nitinol stents and its corrosion behavior were observed. Electropolishing is an effective method for surface treatment, which not only controls the surface state but also helps to produce uniform surface layers. Therefore, to improve the surface quality of nitinol stents, we conducted an electropolishing under various conditions from 30­40 V and 10­30 s as a post heat treatment for nitinol stent manufacturing process. In order to find the optimal surface state of NiTi stents, various electropolished samples were explored using various characterization techniques. Furthermore, the potentiodynamic polarization tests were also performed to determine the corrosion resistance. The electropolished nitinol stents under the condition of 40 V for 10 s exhibited the best corrosion performance as well as surface quality.

On-demand drug release and hyperthermia therapy applications of thermoresponsive poly-(NIPAAm-co-HMAAm)/polyurethane core-shell nanofiber mat on non-vascular nitinol stents.

A functional cover made up of core-shell nanofibers with a unique combination of thermoresponsive polymeric shell and stretchable polymeric core for non-vascular nitinol stents that uses an alternating magnetic field (AMF) to induce heat in the stent for hyperthermia therapy and simultaneously release 5-fluorouracil and/or paclitaxel was designed. Varying the ratios of NIPAAm to HMAAm monomer resulted in different LCST properties for the synthesized copolymer and further utilized for an on-demand drug release. Biocompatibility test using NIH-3T3 fibroblast cells indicates that the composite with drug content is biocompatible and the in-vitro cancer cytotoxicity test using ESO26 and OE21 cancer cells proved that the material shows cancer cytotoxic properties via combination of dual drug and hyperthermia therapy. With this functional material, we propose a tailorable and on-demand drug release with more control that can be employed for a combination drug therapy/single drug therapy combined with hyperthermia therapy for cancer cytotoxicity effect.

Electrospinning Directly Synthesized Porous TiO2 Nanofibers Modified by Graphitic Carbon Nitride Sheets for Enhanced Photocatalytic Degradation Activity under Solar Light Irradiation.

We report a direct approach to the fabrication of a composite made of porous TiO2 nanofibers (NFs) and graphitic carbon nitride (g-C3N4) sheets, by means of an angled two-nozzle electrospinning combined with calcination process. Different wt % amounts of g-C3N4 particles in a polymer solution from one nozzle and TiO2 precursors containing the same polymer solution from another nozzle were electrospun and deposited on the collector. Structural characterizations confirm a well-defined morphology of the TiO2/g-C3N4 composite in which the TiO2 NFs are uniformly attached on the g-C3N4 sheet. This proper attachment of TiO2 NFs on the g-C3N4 sheets occurred during calcination. The prepared composites showed the enhanced photocatalytic activity over the photodegradation of rhodamine B and reactive black 5 under natural sunlight. Here, the synergistic effect between the g-C3N4 sheets and the TiO2 NFs having anisotropic properties enhanced the photogenerated electron-hole pair separation and migration, which was confirmed by the measurement of photoluminescence spectra, cyclic voltammograms, and electrochemical impedance spectra. The direct synthesis approach that is established here for such kinds of sheetlike structure and porous NFs composites could provide new insights for the design of high-performance energy conversion catalysts.

In Situ Synthesis of Antimicrobial Silver Nanoparticles within Antifouling Zwitterionic Hydrogels by Catecholic Redox Chemistry for Wound Healing Application.

A multifunctional hydrogel that combines the dual functionality of both antifouling and antimicrobial capacities holds great potential for many bioapplications. Many approaches and different materials have been employed to synthesize such a material. However, a systematic study, including in vitro and in vivo evaluation, on such a material as wound dressings is highly scarce at present. Herein, we report on a new strategy that uses catecholic chemistry to synthesize antimicrobial silver nanoparticles impregnated into antifouling zwitterionic hydrogels. For this purpose, hydrophobic dopamine methacrylamide monomer (DMA) was mixed in an aqueous solution of sodium tetraborate decahydrate and DMA monomer became soluble after increasing pH to 9 due to the complexation between catechol groups and boron. Then, cross-linking polymerization of zwitterionic monomer was carried out with the solution of the protected dopamine monomer to produce a new hydrogel. When this new hydrogel comes in contact with a silver nitrate solution, silver nanoparticles (AgNPs) are formed in its structure as a result of the redox property of the catechol groups and in the absence of any other external reducing agent. The results obtained from TEM and XRD measurements indicate that AgNPs with diameters of around 20 nm had formed within the networks. FESEM images confirmed that the silver nanoparticles were homogeneously incorporated throughout the hydrogel network, and FTIR spectroscopy demonstrated that the catechol moiety in the polymeric backbone of the hydrogel is responsible for the reduction of silver ions into the AgNPs. Finally, the in vitro and in vivo experiments suggest that these mussel-inspired, antifouling, antibacterial hydrogels have great potential for use in wound healing applications.

Development of cell-laden photopolymerized constructs with bioactive amorphous calcium magnesium phosphate for bone tissue regeneration via 3D bioprinting.

The synthesis of ideal bioceramics to guide the fate of cells and subsequent bone regeneration within the chemical, biological, and physical microenvironment is a challenging long-term task. This study developed amorphous calcium magnesium phosphate (ACMP) bioceramics via a simple co-precipitation method. The role of Mg2+ in the formation of ACMP is investigated using physicochemical and biological characterization at different Ca/Mg molar ratio of the initial reaction solution. Additionally, ACMP bioceramics show superior cytocompatibility and improved osteogenic differentiation of co-cultured MC3T3-E1 cells. Regulation of the microenvironment with Mg2+ can promote early-stage bone regeneration. For this, bioprinting technology is employed to prepare ACMP-modified 3D porous structures. Our hypothesis is that the incorporation of ACMP into methacrylated gelatin (GelMA) bioink can trigger the osteogenic differentiation of encapsulated preosteoblast and stimulate bone regeneration. The cell-laden ACMP composite structures display stable printability and superior cell viability and cell proliferation. Also, constructs loading the appropriate amount of ACMP bioceramic showed significant osteogenic differentiation activity compared to the pure GelMA. We demonstrate that the dissolved Mg2+ cation microenvironment in ACMP-modified composite constructs plays an effective biochemical role, and can regulate cell fate. Our results predict that GelMA/ACMP bioink has significant potential in patient-specific bone tissue regeneration.

Electrochemical technique to develop surface-controlled polyaniline nano-tulips (PANINTs) on PCL-reinforced chitosan functionalized (CS-f-Fe2O3) scaffolds for stimulating osteoporotic bone regeneration.

Bone defects pose significant challenges in orthopedic surgery, often leading to suboptimal outcomes and complications. Addressing these challenges, we employed a three-electrode electrochemical system to fabricate surface-controlled polyaniline nano-tulips (PANINTs) decorated polycaprolactone (PCL) reinforced chitosan functionalized iron oxide nanoparticles (CS-f-Fe2O3) scaffolds. These structures were designed to emulate the natural extracellular matrix (ECM) and promote enhanced osseointegration by establishing a continuous interface between host bone and graft, thereby improving both biological processes and mechanical stability. In vitro experiments demonstrated that PANINTs-PCL/CS-f-Fe2O3 substrates significantly promoted the proliferation, differentiation, and spontaneous outgrowth and extension of MC3T3-E1 cell activity. The nanomaterials exhibited increased cell viability and osteogenic differentiation, as evidenced by elevated expression of bone-related markers such as ALP, ARS, COL-I, RUNX2, and SPP-I, as determined by qRT-PCR. Our findings underscore the regenerative potential of in situ cell culture systems for bone defects, emphasizing the targeted stimulation of essential cell subpopulations to facilitate rapid bone tissue regeneration.