BMSCs-laden mechanically reinforced bioactive sodium alginate composite hydrogel microspheres for minimally invasive bone repair.

Minimally invasive, efficient, and satisfactory treatment for irregular and lacunar bone defects is still a challenge. Alginate hydrogels serve as promising stem cell (SC) delivery systems for bone regeneration but are limited by low cellular viability, poor osteogenic differentiation, and insufficient mechanical support. Herein, we developed a BMSCs-laden mechanically reinforced bioactive sodium alginate composite hydrogel microspheres (BCHMs) system via a microfluidic method that possesses 1) a uniform size and good injectability to meet clinical bone defects with complex shapes, 2) high cellular viability maintenance and further osteogenic induction capacity, and 3) improved mechanical properties. As the main matrix, the sodium alginate hydrogel maintains the high viability of encapsulated BMSCs and efficient substance exchange. Enhanced mechanical properties and osteogenic differentiation of the BCHMs in vitro were observed with xonotlite (Ca6Si6O17(OH)2, CSH) nanowires incorporated. Furthermore, BCHMs with 12.5 % CSH were injected into rat femoral bone defects, and satisfactory in situ regeneration outcomes were observed. Overall, it is believed that BCHMs expand the application of polysaccharide science and provide a promising injectable bone substitute for minimally invasive bone repair.

Preparation and physicochemical characterization of whitlockite/PVA/Gelatin composite for bone tissue regeneration.

This work used a straightforward solvent casting approach to synthesize bone whitlockite (WH) based PVA/Gelatin composites. WH nanoparticles (NPs) were synthesized using the wet precipitation method, followed by their addition into the PVA/Gelatin matrix at concentrations from 1% to 10%. The physicochemical characterization of the prepared PVA/Gelatin/WH composite was carried out using ATR-FTIR, Optical profilometry, a Goniometer, a Universal tensile testing machine (UTM), and scanning electron microscopy (SEM) techniques. The ATR-FTIR analysis confirmed the formation of noncovalent interactions between polymeric chains and WH NPs and the incorporation of WH NPs into the polymer cavities. SEM analysis demonstrated increased surface roughness with the addition of WH NPs, supporting the results obtained through optical profilometry analysis. The mechanical properties of the prepared composite showed an increase in the tensile strength with the addition of WH filler up to 7% loading. The prepared composite has demonstrated an excellent swelling ability and surface wettability. The reported results demonstrate the exceptional potential of the prepared composite for bone tissue regeneration.

Novel cytotoxicity study of strontium (Sr) doped iron oxide (Fe3O4) nanoparticles aided with ibuprofen for drug delivery applications.

This work is aimed at studying the drug delivery applications of iron oxide (Fe3O4) nanoparticles with strontium (Sr) doping with varying molar ratios prepared by the co-precipitation route. The impact of increased strontium content on the particle size and magnetic properties was investigated. The impending of these nanoparticles for drug loading, drug release, and their respective cytotoxicity was also inspected.First, iron oxide nanoparticles were doped with various amounts of strontium, from 0.25, 0.50, and 0.75, to 1 mol using co-precipitation method. These synthesized nanoparticles were characterized by XRD, SEM, EDX, VSM, and FTIR for evaluating crystal structure, phase purity, morphology, composition, magnetic properties, and functional groups, respectively. Drug loading and drug release properties were determined using UV-vis spectroscopy, whereas MTT assay evaluated cytotoxicity. Colloidal stability was assessed using zeta potential in PBS solution.The findings confirmed the successful doping of iron oxide with strontium via XRD and EDX. SEM results confirmed spherical morphology for all and needle-like structure for 1 mol strontium doped sample. For VSM results, a single domain structure was established. It was also observed that the drug encapsulation efficiency increases with increased strontium content. Cytotoxicity results by MTT assay revealed increased cytotoxicity with increasing nanoparticle concentration, and ibuprofen-loaded nanoparticles showed higher cytotoxicity than un-loaded nanoparticles at the same concentration. Zeta potential results showed colloidal stability of iron oxide nanoparticles increased by the addition of strontium.This study provided predominantly comparison of the cytotoxicity of ibuprofen-loaded and non-loaded nanoparticles on Hep-2 cancer cells at similar concentrations for the first time for both Fe3O4 particles and Sr-doped Fe3O4 nanoparticles and enclosed the impact of increasing Sr doping content on Fe3O4 nanoparticles.

Advances in Gelatin Bioinks to Optimize Bioprinted Cell Functions.

Gelatin is a widely utilized bioprinting biomaterial due to its cell-adhesive and enzymatically cleavable properties, which improve cell adhesion and growth. Gelatin is often covalently cross-linked to stabilize bioprinted structures, yet the covalently cross-linked matrix is unable to recapitulate the dynamic microenvironment of the natural extracellular matrix (ECM), thereby limiting the functions of bioprinted cells. To some extent, a double network bioink can provide a more ECM-mimetic, bioprinted niche for cell growth. More recently, gelatin matrices are being designed using reversible cross-linking methods that can emulate the dynamic mechanical properties of the ECM. This review analyzes the progress in developing gelatin bioink formulations for 3D cell culture, and critically analyzes the bioprinting and cross-linking techniques, with a focus on strategies to optimize the functions of bioprinted cells. This review discusses new cross-linking chemistries that recapitulate the viscoelastic, stress-relaxing microenvironment of the ECM, and enable advanced cell functions, yet are less explored in engineering the gelatin bioink. Finally, this work presents the perspective on the areas of future research and argues that the next generation of gelatin bioinks should be designed by considering cell-matrix interactions, and bioprinted constructs should be validated against currently established 3D cell culture standards to achieve improved therapeutic outcomes.

Emerging Methods in Biosensing of Immunoglobin G-A Review.

Human antibodies are produced due to the activation of immune system components upon exposure to an external agent or antigen. Human antibody G, or immunoglobin G (IgG), accounts for 75% of total serum antibody content. IgG controls several infections by eradicating disease-causing pathogens from the body through complementary interactions with toxins. Additionally, IgG is an important diagnostic tool for certain pathological conditions, such as autoimmune hepatitis, hepatitis B virus (HBV), chickenpox and MMR (measles, mumps, and rubella), and coronavirus-induced disease 19 (COVID-19). As an important biomarker, IgG has sparked interest in conducting research to produce robust, sensitive, selective, and economical biosensors for its detection. To date, researchers have used different strategies and explored various materials from macro- to nanoscale to be used in IgG biosensing. In this review, emerging biosensors for IgG detection have been reviewed along with their detection limits, especially electrochemical biosensors that, when coupled with nanomaterials, can help to achieve the characteristics of a reliable IgG biosensor. Furthermore, this review can assist scientists in developing strategies for future research not only for IgG biosensors but also for the development of other biosensing systems for diverse targets.

Novel Method for the Production of Titanium Foams to Reduce Stress Shielding in Implants.

Titanium foams have potential applications in orthopedic and dental implants because of their low elastic modulus and good bone in-growth properties. In the present study, a novel method for the preparation of three-dimensional interconnected microporous titanium foams has been developed. This method is based on the insertion of a filler metal into the titanium metal by arc melting, followed by its removal by an electrochemical dealloying process for the development of foams. Complete removal of the filler metal by the electrochemical dealloying process was confirmed by an X-ray diffractometry (XRD) analysis, whereas scanning electron microscopy (SEM) analysis of the developed foams showed the development of interconnected porosity. Ti foams with different levels of porosities were successfully developed by varying the amount of the filler metal. Mechanical and thermal characterizations of the developed foams were carried out using compression testing and laser flash apparatus, respectively. The yield strength and elastic modulus of the developed foams were found to decrease by increasing the volume fraction of pores. The elastic modulus of the developed titanium foams (15.5-36 GPa) was found to be closer to that of human bones, whereas their yield strength (147-170 MPa) remained higher than that of human bones. It is therefore believed that the developed Ti foams can help in reducing the problem of stress shielding observed in orthopedic implants. The thermal diffusivity of the developed foams (4.3-0.69 mm2/s) was found to be very close to that of human dentine.

Effects of L-valine in layer diets containing 0.72% isoleucine.

In a previous study with LSL-LITE layers (-23 to 30-week-old), isoleucine at 0.72% and 0.84% produced values for FCR at 1.45 and 1.44, respectively and shared significance with 0.78% isoleucine (1.49). Considering that FCR is an important standard in the poultry industry due to the cost for adding feed ingredients such as synthetic amino acids and the low FCR of 1.45, 0.72% isoleucine was chosen for further study with LSL-LITE layers (n = 490 at 33- to 40-week-old) to determine effects on production and egg quality. The study included 7 diets (2730 Kcal kg metabolizable energy and constant isoleucine at 0.72%) containing varying quantities of valine [0.72 (Control), 0.75, 0.78, 0.81, 0.84, 0.87 or 0.90%] x 7 replicates x 10 hens/replicate. Significance at P ≤ 0.05 and P < 0.10 was determined. Level and week were significant for feed intake, egg production, and FCR; the interaction of level x week (L*W) was significant for feed intake and FCR. An isoleucine:valine of 1.233 corresponding to 0.72% isoleucine and 0.87% valine produced the lowest FCR of 1.30 (a 2.26% decrease compared to the Control at 1.33 ± 0.04). All measurements for external egg quality, except shape index and eggshell thickness, were significant for level. Week was significant for all parameters except shell thickness; L*W was significant for external quality measurements except shape index and shell thickness. Level, week, and L*W were significant for internal egg quality measurements. Serum protein and H1 titer were significant for level. Various production, egg quality, and biochemical measurements were significantly different from the control (0.72% isoleucine and 0.72% valine) at 0.81 to 0.87% valine. Findings of this study will aid researchers and commercial producers in narrowing the range of isoleucine, valine, and leucine needed for effects on particular parameters. Knowledge gained from this and others studies will eventually lead to an understanding of synergistic and antagonistic effects of branched chain amino acids in feed for various genetic types of layers throughout their productive lifetime.

Liquid Crystal Based Binding Assay for Detecting HIV-1 Surface Glycoprotein.

Surface protein gp-120 of HIV-1 virus plays an important role in the infection of HIV-1, but detection of gp-120 during the early stage of infection is very difficult. Herein, we report a binding bioassay based on an RNA aptamer B40t77, which binds specifically to gp-120. The bioassay is built upon a hydrophobic glass slide with surface immobilized gp-120. When the glass surface is incubated in a solution containing B40t77, the aptamer is able to bind to gp-120 specifically and remove it from the surface after a short incubation time of 30 min. The result of the binding event can be amplified by using liquid crystal (LC) into optical signals in the final step. By using this bioassay, we are able to detect as low as 1 µg/ml of gp-120 with high specificity within 30 min. No response is obtained when gp-120 is replaced by other protein such as bovine serum albumin (BSA). This is the first qualitative bioassay which provides a simple way for the detection of gp-120 with the naked eye. The assay is robust, low-cost and does not require additional labeling. Thus, the bioassay is potentially useful for the early detection of HIV-1 in resources-limited regions.

Detection of lead in soil implying sample heating and laser-induced breakdown spectroscopy.

The emission line intensities enhancement and sensitivity of laser-induced breakdown spectroscopy (LIBS) has been a subject of great interest for the last several years to improve the detection of the trace elements in soil and other environmental samples. Among several other methodologies, LIBS of the heated targets is emerging as one of the effective techniques to achieve the objective. We have investigated the effect of target heating (room temperature, 100°C, and 200°C) on the emission enhancement and plasma parameters of the laser-produced plasma on the soil sample containing 80 ppm lead. In addition, the limit of detection (LOD) of lead in soil has been determined at a fixed target temperature (200°C) and with varying lead concentration (20 ppm, 80 ppm, and 100 ppm) in the soil samples. With increasing the target temperature, not only do the emission line intensities, the excitation temperature, and electron number density increase, but also the spectral lines of Pb emerge, which were absent in the soil spectra recorded at room temperature. The limit of detection of lead in LIBS of the heated soil target has been determined as 3.8 ppm. This study reveals the potential application of the LIBS of a pre-heated target for the detection of lead with an improved LOD in the environmental sample.

Preparation of PBS/PLLA/HAP Composites by the Solution Casting Method: Mechanical Properties and Biocompatibility.

The use of biodegradable polymeric scaffolds for tissue regeneration is becoming a common practice in the clinic. Therefore, an inclined trend is developing with regards to improving the mechanical properties of these scaffolds. Here, we aim to improve the mechanical properties of poly (butylene succinate) (PBS)/poly (l-lactic acid) (PLLA) blends by incorporating hydroxyapatite nanoparticles (HAP) in the blends to form composites. PBS/PLLA = 100/0, 95/5, 90/10, 85/15, and 0/100 wt% blends, along-with the loadings of a few mg of HAPs, were prepared using the solution casting method. A scanning electron microscope showed the voids and droplets, indicating the immiscibility of blends. Due to this immiscibility, the tensile strength values of the blends were found to be in between that of pure PBS (42.85 MPa) and pure PLLA (31.39 MPa). HAPs act as a compatibilizer by incorporating themselves in the voids and spaces caused by the immiscibility, thus increasing the overall tensile strength of the resulting composite to a certain extent, e.g., the tensile strength of PBS/PLLA = 95/5 loaded with 50 mg HAPs was found to be 51.16 MPa. The structural analysis employing the X-ray diffraction (XRD) patterns confirmed the formation of polymer blends and composites. The contact angle analysis showed that the addition of HAPs increased the hydrophilicity of the resulting composites. Selective samples were investigated based on mechanical properties to see if the blends and composites are biocompatible. The obtained results showed that all of the samples with better mechanical properties demonstrated good biocompatibility. This indicates the effectiveness of scaffolds for tissue regeneration.

Synthesis, Characterization and Process Optimization of Bone Whitlockite.

Whitlockite, being the second most abundant bio-mineral in living bone, finds huge applications in tissue regeneration and implants and its synthesis into its pure form has remained a challenge. Although precipitation of whitlockite phase has been reported recently in many publications, effects of various parameters to control such phase as well as conditions for the bulk preparation of this extremely important bio-mineral have not been investigated so far. In this work, we report the precipitation of pure whitlockite phase using common precursors. As reported in the literature, whitlockite is stable in a narrow pH range, therefore; optimization of pH for the stabilization of whitlockite phase has been investigated. Additionally, in order to narrow down the optimum conditions for the whitlockite precipitation, effect of temperature and heating conditions has also been studied. The obtained solids were characterized using powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, scanning electron microscopy (SEM) and thermogravimetric analysis (TGA). From PXRD analysis, it was observed that heating the precursor's mixture at 100 °C with subsequent aging at the optimized pH resulted in the precipitation of pure whitlockite phase. These results were further confirmed by TGA, SEM and Raman spectroscopy analysis and it was confirmed that the conditions reported here favor whitlockite precipitation without formation of any secondary phase. These reaction conditions were further confirmed by changing all the parameters like aging, heating time, feed rate of precursors one by one. From PXRD analysis of these samples, it was concluded that not only pH but temperature, heating time, aging time and feed rate effect simultaneously on the precipitation of pure whitlockite phase and a subtle change in any of these parameters could lead to the formation of undesired stable secondary calcium phosphate phases.

Elemental composition analysis of granite rocks using LIBS and LA-TOF-MS.

Qualitative and quantitative analysis of the raw granite rocks acquired from deposits in the Hunza district, Gilgit area of Pakistan were studied using laser-induced breakdown spectroscopy (LIBS) and laser ablation time-of-flight mass spectrometry (LA-TOF-MS). The optical emission spectra of the granite rocks used in artificial jewelry and for flooring tiles show the emission lines of (Si, Ca, K, Fe, Mg, Al, Na, and Li) and (Si, Ca, K, Fe, Mg, Al, Ti, Na, Ba, and Li), respectively. The mass spectra of these granite rocks were also studied with the LA-TOF-MS, revealing analogous elemental compositions. The results obtained using LIBS and LA-TOF-MS for the raw granite samples display their ability as powerful and complementary tools for the compositional analysis of the geological samples.

UV-irradiated parylene surfaces for proliferation and differentiation of PC-12 cells.

PC-12 cells originate from neuroblastic cells, which have an ability to differentiate into neuronlike cells. In this work, the purpose was to estimate the influence of microenvironments on cell attachment and neuritogenesis capacity of PC-12 cells on parylene-N and parylene-C films with and without ultraviolet (UV) light treatment. The estimate of total cell number after incubation for 72h, the ratio of adherent to suspended cells, counting of neurite outgrowths on parylene-N or parylene-C films after UV exposure suggested that these films were suitable for proliferation as well as differentiation of PC-12 cells. The differences in surface properties of parylene-N and parylene-C films with and without UV exposure were analyzed by contact angle measurement, Fourier-transform infrared spectrometry, and X-ray photoelectron spectroscopy. According to these analyses, introduction of oxygen-related chemical functional groups was presumed to result in increased hydrophilicity and efficiency of protein immobilization on parylene-N and parylene-C films after UV treatment. According to fluorescent staining, western blotting, and cell cycle analysis, UV-treated parylene-C and parylene-N films appear to effectively facilitate simultaneous proliferation and differentiation of PC-12 cells with neurite outgrowth.