Combining Copper and Zinc into a Biosensor for Anti-Chemoresistance and Achieving Osteosarcoma Therapeutic Efficacy.

Due to its built-up chemoresistance after prolonged usage, the demand for replacing platinum in metal-based drugs (MBD) is rising. The first MBD approved by the FDA for cancer therapy was cisplatin in 1978. Even after nearly four and a half decades of trials, there has been no significant improvement in osteosarcoma (OS) therapy. In fact, many MBD have been developed, but the chemoresistance problem raised by platinum remains unresolved. This motivates us to elucidate the possibilities of the copper and zinc (CuZn) combination to replace platinum in MBD. Thus, the anti-chemoresistance properties of CuZn and their physiological functions for OS therapy are highlighted. Herein, we summarise their chelators, main organic solvents, and ligand functions in their structures that are involved in anti-chemoresistance properties. Through this review, it is rational to discuss their ligands' roles as biosensors in drug delivery systems. Hereafter, an in-depth understanding of their redox and photoactive function relationships is provided. The disadvantage is that the other functions of biosensors cannot be elaborated on here. As a result, this review is being developed, which is expected to intensify OS drugs with higher cure rates. Nonetheless, this advancement intends to solve the major chemoresistance obstacle towards clinical efficacy.

Composing On-Program Triggers and On-Demand Stimuli into Biosensor Drug Carriers in Drug Delivery Systems for Programmable Arthritis Therapy.

Medication in arthritis therapies is complex because the inflammatory progression of rheumatoid arthritis (RA) and osteoarthritis (OA) is intertwined and influenced by one another. To address this problem, drug delivery systems (DDS) are composed of four independent exogenous triggers and four dependent endogenous stimuli that are controlled on program and induced on demand, respectively. However, the relationships between the mechanisms of endogenous stimuli and exogenous triggers with pathological alterations remain unclear, which results in a major obstacle in terms of clinical translation. Thus, the rationale for designing a guidance system for these mechanisms via their key irritant biosensors is in high demand. Many approaches have been applied, although successful clinical translations are still rare. Through this review, the status quo in historical development is highlighted in order to discuss the unsolved clinical difficulties such as infiltration, efficacy, drug clearance, and target localisation. Herein, we summarise and discuss the rational compositions of exogenous triggers and endogenous stimuli for programmable therapy. This advanced active pharmaceutical ingredient (API) implanted dose allows for several releases by remote controls for endogenous stimuli during lesion infections. This solves the multiple implantation and local toxic accumulation problems by using these flexible desired releases at the specified sites for arthritis therapies.

CuZn Complex Used in Electrical Biosensors for Drug Delivery Systems.

This paper is to discuss the potential of using CuZn in an electrical biosensor drug carrier for drug delivery systems. CuZn is the main semiconductor ingredient that has great promise as an electrochemical detector to trigger releases of active pharmaceutical ingredients (API). This CuZn biosensor is produced with a green metal of frameworks, which is an anion node in conductive polymers linked by bioactive ligands using metal-polymerisation technology. The studies of Cu, Zn, and their oxides are highlighted by their electrochemical performance as electrical biosensors to electrically trigger API. The three main problems, which are glucose oxidisation, binding affinity, and toxicity, are highlighted, and their solutions are given. Moreover, their biocompatibilities, therapeutic efficacies, and drug delivery efficiencies are discussed with details given. Our three previous investigations of CuZn found results similar to those of other authors' in terms of multiphases, polymerisation, and structure. This affirms that our research is on the right track, especially that related to green synthesis using plant extract, CuZn as a nanochip electric biosensor, and bioactive ligands to bind API, which are limited to the innermost circle of the non-enzymatic glucose sensor category.

SCOBY-based bacterial cellulose as free standing electrodes for safer, greener and cleaner energy storage technology.

Bacterial Cellulose (BC) derived from local market or symbiotic culture of bacteria and yeast (SCOBY) was employed as the polymer matrix for hydroxyl multi-walled carbon nanotube (MWCNT-OH)-based electrochemical double-layer capacitor (EDLC). Chitosan (CS)-sodium iodide (NaI)-glycerol (Gly) electrolyte systems were used as the polymer electrolyte. CS-NaI-Gly electrolyte possesses conductivity, potential stability and ionic transference number of (1.20 ± 0.26) × 10-3 S cm-2, 2.5 V and 0.99, respectively. For the electrodes, MWCNT-OH was observed to be well dispersed in the matrix of BC which was obtained via FESEM analysis. The inclusion of MWCNT-OH reduced the crystallinity of the BC polymeric structure. From EIS measurement, it was verified that the presence of MWCNT-OH decreased the electron transfer resistance of BC-based electrodes. Cyclic voltammetry (CV) showed that the shape of the CV plots changed to a rectangular-like shape plot as more MWCNT were added, thus verifying the capacitive behavior. Various amount of MWCNT-OH was used in the fabrication of the EDLC where it was discovered that more MWCNT-OH leads to a better EDLC performance. The EDLC was tested for 5000 complete charge-discharge cycles. The optimum performance of this low voltage EDLC was obtained with 0.1 g MWCNT where the average specific capacitance was 8.80 F g-1. The maximum power and energy density of the fabricated EDLC were 300 W kg-1 and 1.6 W h kg-1, respectively.

Multifunction Web-like Polymeric Network Bacterial Cellulose Derived from SCOBY as Both Electrodes and Electrolytes for Pliable and Low-Cost Supercapacitor.

In this work, bacterial cellulose (BC)-based polymer derived from a symbiotic culture of bacteria and yeast (SCOBY) are optimized as both electrodes and electrolytes to fabricate a flexible and free-standing supercapacitor. BC is a multifunction and versatile polymer. Montmorillonite (MMT) and sodium bromide (NaBr) are used to improve mechanical strength and as the ionic source, respectively. From XRD analysis, it is found that the addition of MMT and NaBr has reduced the crystallinity of the electrolyte. Most interaction within the electrolyte happens in the region of the OH band, as verified using FTIR analysis. A maximum room temperature conductivity of (1.09 ± 0.02) × 10-3 S/cm is achieved with 30 wt.% NaBr. The highest conducting SCOBY-based electrolytes have a decompose voltage and ionic transference number of 1.48 V and 0.97, respectively. The multiwalled carbon nanotube is employed as the active material held by the fibrous network of BC. Cyclic voltammetry shows a rectangular shape CV plot with the absence of a redox peak. The supercapacitor is charged and discharged in a zig-zag-shaped Perspex plate for 1000 cycles with a decent performance.

Structural Strength Analyses for Low Brass Filler Biomaterial with Anti-Trauma Effects in Articular Cartilage Scaffold Design.

The existing harder biomaterial does not protect the tissue cells with blunt-force trauma effects, making it a poor choice for the articular cartilage scaffold design. Despite the traditional mechanical strengths, this study aims to discover alternative structural strengths for the scaffold supports. The metallic filler polymer reinforced method was used to fabricate the test specimen, either low brass (Cu80Zn20) or titanium dioxide filler, with composition weight percentages (wt.%) of 0, 2, 5, 15, and 30 in polyester urethane adhesive. The specimens were investigated for tensile, flexural, field emission scanning electron microscopy (FESEM), and X-ray diffraction (XRD) tests. The tensile and flexural test results increased with wt.%, but there were higher values for low brass filler specimens. The tensile strength curves were extended to discover an additional tensile strength occurring before 83% wt.%. The higher flexural stress was because of the Cu solvent and Zn solute substituting each other randomly. The FESEM micrograph showed a cubo-octahedron shaped structure that was similar to the AuCu3 structure class. The XRD pattern showed two prominent peaks of 2θ of 42.6° (110) and 49.7° (200) with d-spacings of 1.138 Å and 1.010 Å, respectively, that indicated the typical face-centred cubic superlattice structure with Cu and Zn atoms. Compared to the copper, zinc, and cart brass, the low brass indicated these superlattice structures had ordered-disordered transitional states. As a result, this additional strength was created by the superlattice structure and ordered-disordered transitional states. This innovative strength has the potential to develop into an anti-trauma biomaterial for osteoarthritic patients.

Magnetic exposure using Samarium Cobalt (SmCO5) increased proliferation and stemness of human Umbilical Cord Mesenchymal Stem Cells (hUC-MSCs).

Despite the extensive reports on the potential hazard of magnetic field (MF) exposures on humans, there are also concurrently reported on the improved proliferative property of stem cells at optimum exposure. However, the effect on mesenchymal stem cells (MSCs) remains unknown. Therefore, we aimed to investigate the impact of induced static MF (SMF) on human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) using Samarium Cobalt (SmCO5). At passage 3, hUC-MSCs (1 × 104) were exposed to 21.6 mT SMF by a direct exposure (DE) showed a significantly higher cell count (p < 0.05) in the growth kinetics assays with the shortest population doubling time relative to indirect exposure and negative control. The DE group was committed into the cell cycle with increased S phase (55.18 ± 1.38%) and G2/M phase (21.75 ± 1.38%) relative to the NC group [S-phase (13.54 ± 2.73%); G2/M phase (8.36 ± 0.28%)]. Although no significant changes were observed in the immunophenotype, the DE group showed an elevated expression of pluripotency-associated markers (OCT4, SOX2, NANOG, and REX1). These results suggest that the MFs could potentially induce proliferation of MSCs, a promising approach to promote stem cells propagation for clinical therapy and research without compromising the stemness of hUC-MSCs.

Numerical Simulation Study on Relationship between the Fracture Mechanisms and Residual Membrane Stresses of Metallic Material.

The distribution and dissipation energies in fracture mechanisms were a critical challenge to derive, especially for this ultra-thin sample. The membrane failure, which is the end of the fracture mechanisms, is a result of the cone wave reflections from the backend membrane boundaries. These reflections delay the failure processes due to the shock impacts. To compare these results with the experimental work, a numerical simulation was conducted for these processes. The cylinder-shaped rigid projectile was impacted using a frictionless Lagrange solver. The target was a cartridge brass circle plate clamped at its perimeter, and its zone was refined to a ten-times higher meshing density for better analysis. The erosion and cut-off controls involved a zero-gap interaction condition and an instantaneous geometric erosion strain of 200%. Due to the maximum projectile velocity of 382 m/s having the slowest perforation, the target thickness was found to be 5.5 mm. The fracture mechanism phenomena, such as tensile, compressive, through-thickness, and growth in-plane delamination, propagating delamination, and local punch shear waves were observed. After deducting tensile and flexural strengths from the last experiment, a total residual membrane stress of 650 MPa was found. This result indicated a relationship between the fracture mechanisms and residual membrane stresses of metallic material.

Structural Characterization Analyses of Low Brass Filler Biomaterial for Hard Tissue Implanted Scaffold Applications.

A biomaterial was created for hard tissue implanted scaffolds as a translational therapeutic approach. The existing biomaterials containing titanium dioxide filler posed a risk of oxygen gas vacancy. This will block the canaliculars, leading to a limit on the nutrient fluid supply. To overcome this problem, low brass was used as an alternative filler to eliminate the gas vacancy. Low brass with composition percentages of 0%, 2%, 5%, 15%, and 30% was filled into the polyester urethane liquidusing the metallic filler polymer reinforced method. The structural characterizations of the low brass filler biomaterial were investigated by Field Emission Scanning Electron Microscopy. The results showed the surface membrane strength was higher than the side and cross-section. The composition shapes found were hexagon for polyester urethane and peanut for low brass. Low brass stabilised polyester urethane in biomaterials by the formation of two 5-ringed tetrahedral crystal structures. The average pore diameter was 308.9 nm, which is suitable for articular cartilage cells. The pore distribution was quite dispersed, and its curve had a linear relationship between area and diameter, suggestive of the sphere-shaped pores. The average porosities were different between using FESEM results of 6.04% and the calculated result of 3.28%. In conclusion, this biomaterial had a higher surface membrane strength and rather homogeneous dispersed pore structures.

Electromagnetic field exposure as a plausible approach to enhance the proliferation and differentiation of mesenchymal stem cells in clinically relevant scenarios.

Mesenchymal stem/stromal cell (MSC)|-based therapy has been regarded as one of the most revolutionary breakthroughs in the history of modern medicine owing to its myriad of immunoregulatory and regenerative properties. With the rapid progress in the fields of osteo- and musculoskeletal therapies, the demand for MSC-based treatment modalities is becoming increasingly prominent. In this endeavor, researchers around the world have devised new and innovative techniques to support the proliferation of MSCs while minimizing the loss of hallmark features of stem cells. One such example is electromagnetic field (EMF) exposure, which is an alternative approach with promising potential. In this review, we present a critical discourse on the efficiency, practicability, and limitations of some of the relevant methods, with insurmountable evidence backing the implementation of EMF as a feasible strategy for the clinically relevant expansion of MSCs.

An electrochemical sensor based on gold nanoparticles-functionalized reduced graphene oxide screen printed electrode for the detection of pyocyanin biomarker in Pseudomonas aeruginosa infection.

Multidrug resistant Pseudomonas aeruginosa (P. aeruginosa) is known to be a problematic bacterium for being a major cause of opportunistic and nosocomial infections. In this study, reduced graphene oxide decorated with gold nanoparticles (AuNPs/rGO) was utilized as a new sensing material for a fast and direct electrochemical detection of pyocyanin as a biomarker of P. aeruginosa infections. Under optimal condition, the developed electrochemical pyocyanin sensor exhibited a good linear range for the determination of pyocyanin in phosphate-buffered saline (PBS), human saliva and urine at a clinically relevant concentration range of 1-100 µM, achieving a detection limit of 0.27 µM, 1.34 µM, and 2.3 µM, respectively. Our developed sensor demonstrated good selectivity towards pyocyanin in the presence of interfering molecule such as ascorbic acid, uric acid, NADH, glucose, and acetylsalicylic acid, which are commonly found in human fluids. Furthermore, the developed sensor was able to discriminate the signal with and without the presence of pyocyanin directly in P. aeruginosa culture. This proposed technique demonstrates its potential application in monitoring the presence of P. aeruginosa infection in patients.

Improved Drug Delivery System for Cancer Treatment by D-Glucose Conjugation with Eugenol From Natural Product.

INTRODUCTION: Bioconjugations are swiftly progressing and are being applied to solve several limitations of conventional Drug Delivery Systems (DDS) such as lack of water solubility, non-specific, and poor bioavailability. The main goals of DDS are to achieve greater drug effectiveness and minimize toxicity to the healthy tissues. OBJECTIVES AND METHODS: In this study, D-glucose was conjugated with eugenol to target the cancer cells. To identify the implication of the anticancer effect, osteosarcoma (K7M2) cells were cultured and the anti-proliferative effect was performed using MTT [3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide assay] test in order to evaluate the viability and toxicity on cells with various concentrations of eugenol and D-glucose-eugenol conjugate in 24-hour incubation. RESULTS: It was found that, the successful confirmation of the conjugation between D-glucose and eugenol was obtained by 1H NMR spectroscopy. MTT assay showed inhibitory concentration (IC50 value) of D-glucose-eugenol was at 96.2 µg/ml and the decreased of osteosarcoma cell survival was 48%. Conclucion: These findings strongly indicate that K7M2 cells would be affected by toxicity of Dglucose- eugenol. Therefore, the present study suggests that D-glucose-eugenol has high potential to act as an anti-proliferative agent who may promise a new modality or approach as the drug delivery treatment for cancer or chemotherapeutic agent.

Boron Nitride Doped Polyhydroxyalkanoate/Chitosan Nanocomposite for Antibacterial and Biological Applications.

The present research focused on the fabrication of biocompatible polyhydroxyalkanoate, chitosan, and hexagonal boron nitride incorporated (PHA/Ch-hBN) nanocomposites through a simple solvent casting technique. The fabricated nanocomposites were comprehensively characterized by Fourier transform infrared spectroscope (FT-IR), field emission scanning electroscope (FESEM), and elemental mapping and thermogravimetric analysis (TGA). The antibacterial activity of nanocomposites were investigated through time-kill method against multi drug resistant (MDR) microbes such as methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli) K1 strains. In addition, nanocomposites have examined for their host cytotoxicity abilities using a Lactate dehydrogenase (LDH) assay against spontaneously immortalized human keratinocytes (HaCaT) cell lines. The results demonstrated highly significant antibacterial activity against MDR organisms and also significant cell viability as compared to the positive control. The fabricated PHA/Ch-hBN nanocomposite demonstrated effective antimicrobial and biocompatibility properties that would feasibly suit antibacterial and biomedical applications.

Application of stem cells in tissue engineering for defense medicine.

The dynamic nature of modern warfare, including threats and injuries faced by soldiers, necessitates the development of countermeasures that address a wide variety of injuries. Tissue engineering has emerged as a field with the potential to provide contemporary solutions. In this review, discussions focus on the applications of stem cells in tissue engineering to address health risks frequently faced by combatants at war. Human development depends intimately on stem cells, the mysterious precursor to every kind of cell in the body that, with proper instruction, can grow and differentiate into any new tissue or organ. Recent reports have suggested the greater therapeutic effects of the anti-inflammatory, trophic, paracrine and immune-modulatory functions associated with these cells, which induce them to restore normal healing and tissue regeneration by modulating immune reactions, regulating inflammation, and suppressing fibrosis. Therefore, the use of stem cells holds significant promise for the treatment of many battlefield injuries and their complications. These applications include the treatment of injuries to the skin, sensory organs, nervous system tissues, the musculoskeletal system, circulatory/pulmonary tissues and genitals/testicles and of acute radiation syndrome and the development of novel biosensors. The new research developments in these areas suggest that solutions are being developed to reduce critical consequences of wounds and exposures suffered in warfare. Current military applications of stem cell-based therapies are already saving the lives of soldiers who would have died in previous conflicts. Injuries that would have resulted in deaths previously now result in wounds today; similarly, today's permanent wounds may be reduced to tomorrow's bad memories with further advances in stem cell-based therapies.

Individual evaluation of cardiac marker expression and self-beating during cardiac differentiation of P19CL6 cells on different culture substrates.

Cell-based therapies using self-beating cardiomyocytes have been attracting great attention for use in cardiac regeneration, although an effective procedure to improve cardiac differentiation and self-beating induction is required. The purpose of this study is to clarify the effect of the culture substrate on cardiac maturation by separately evaluating the cardiac differentiation step and the beating induction step in vitro. To this end, the well-studied cardiomyocyte-like progenitor cell line P19CL6 and neonatal cardiomyocytes (NCMs) were selected and cultured on substrates coated with collagen type I (Col-I), gelatin (Gel), fibronectin (FN), or poly-l-lysine (PLL). It was found that the cardiac differentiation step, which was assessed using cardiac marker gene expression (GATA-binding protein 4 (GATA4), myocyte-specific enhancer factor 2D (MEF2D), and hyperpolarization-activated cyclic nucleotide-gated potassium channel 4 (HCN4)) in the P19CL6 embryonal carcinoma cells, was greatly enhanced on Col-I, Gel, and PLL. In contrast, the spontaneous beating step, which was directly assessed by counting the beating colonies and measuring contractile protein gene expression (α-myosin heavy chain (α-MHC), troponin C type 1 (TnC1), and troponin T type 2 (TnT2)) in the rat NCMs, was enhanced on the FN and PLL surfaces. In the present study, for the first time, it was found that PLL enhances both the cardiac differentiation and the beating induction steps of cardiac maturation, which can aid in preparing beating cardiomyocytes for regenerative medicine. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1166-1174, 2017.

A suspension induction for myocardial differentiation of rat mesenchymal stem cells on various extracellular matrix proteins.

The microenvironment of bone marrow-derived mesenchymal stem cells (MSCs) strictly regulates their differentiation. In this study, we have developed a new suspension induction method for myocardial differentiation of bone marrow-derived rat MSCs (rMSCs) in vitro on various extracellular matrix (ECM) proteins. Myocardial differentiation of rMSCs was induced with a conventional monolayer method and our suspension method. In our suspension induction, a cell suspension was treated with the medium in the presence of an inducer, incubated for 2 h under a suspension conditions, and moved to a monolayer culture on gelatin-coated, collagen type I-coated, fibronectin-coated, or polystyrene dishes until the total induction time was 24 h. We evaluated the myocardial differentiation by counting the number of colonies of beating cells, performing immunohistochemical staining, and measuring the expression of cardiac-specific gene mRNA using real-time quantitative polymerase chain reaction. We found that rMSCs induced with the conventional monolayer method did not differentiate efficiently, whereas beating cell colonies were found on ECM-coated dishes of suspension-induced cells, after 3 weeks of culture, especially on gelatin-coated dishes. The beating cells were positively stained with anti-troponin T-C antibody and expressed specific cardiac markers. In conclusion, these results demonstrated that the suspension induction followed by subsequent culture on gelatin ECM substrates is a promising method for differentiating rMSCs into cardiomyocytes in vitro.

Beating behavior of primary neonatal cardiomyocytes and cardiac-differentiated P19.CL6 cells on different extracellular matrix components.

Stem cell-based therapy in cardiac tissue engineering is an emerging field that shows great potential for treating heart diseases. However, even preliminary issues, such as the ideal niche for cardiomyocytes, have not been clarified yet. In the present study, the effects of extracellular matrix (ECM) components on the beating duration of neonatal rat cardiomyocytes (RCMs) and on the cardiac differentiation of P19.CL6 carcinoma stem cells were studied. RCMs were cultured on gelatin-, fibronectin-, and collagen type I-coated dishes and on noncoated polystyrene dishes, and their beating rate, beating duration, and cardiac gene expression were evaluated. The beating period and the expression of troponin T type-2 (TNNT2) and troponin C type-1 (TNNC1) of cardiomyocytes cultured on gelatin-coated dishes were longer and higher than for those on dishes with other coatings. For the cardiac differentiation of P19.CL6 cells, troponin T type-2 expression on gelatin- and fibronectin-coated dishes was five times that on collagen type I-coated dishes or polystyrene dishes 11 days after induction. These results indicate that a gelatin-coated surface has a high ability not only to maintain the cardiac phenotype but also to enhance cardiac differentiation.

Preservation of porcine hepatocytes in three-dimensional bioreactor at room temperature using epigallocatechin-3-gallate.

A bioartificial liver (BAL) assist system employing a three-dimensional (3D) bioreactor has been studied as a temporary support in liver failure. In the present study, a novel preservation method of primary cultured porcine hepatocytes in monolayer and 3D culture systems was studied. Epigallocatechin-3-gallate (EGCG), which has recently been found to have various bioactivities, was selected as a key compound for hepatocyte preservation. Hepatocytes isolated from porcine liver using the collagenase perfusion method were pre-cultured for 6 days, preserved at room temperature in the presence of EGCG at various concentrations for 4 days, and post-cultured in normal medium for another 6 days. In the monolayer culture, only albumin production rate was fully recovered after preservation when EGCG concentration was high (0.25mg/mL). In contrast, albumin production and ammonium metabolism in the 3D bioreactor under the same condition recovered to 72+/-16% and 98+/-32%, respectively, of levels before preservation. These results indicate that hepatocytes can be preserved in the presence of 0.25mg/mL of EGCG at room temperature, especially in a 3D culture system, which is promising technology for BAL preparation.