Conjugation of bone grafts with NO-delivery dinitrosyl iron complexes promotes synergistic osteogenesis and angiogenesis in rat calvaria bone defects.

Craniofacial/jawbone deformities remain a significant clinical challenge in restoring facial/dental functions and esthetics. Despite the reported therapeutics for clinical bone tissue regeneration, the bioavailability issue of autografts and limited regeneration efficacy of xenografts/synthetic bone substitutes, however, inspire continued efforts towards functional conjugation and improvement of bioactive bone graft materials. Regarding the potential of nitric oxide (NO) in tissue engineering, herein, functional conjugation of NO-delivery dinitrosyl iron complex (DNIC) and osteoconductive bone graft materials was performed to optimize the spatiotemporal control over the delivery of NO and to activate synergistic osteogenesis and angiogenesis in rat calvaria bone defects. Among three types of biomimetic DNICs, [Fe2(µ-SCH2CH2COOH)2(NO)4] (DNIC-COOH) features a steady kinetics for cellular uptake by MC3T3-E1 osteoblast cells followed by intracellular assembly of protein-bound DNICs and release of NO. This steady kinetics for intracellular delivery of NO by DNIC-COOH rationalizes its biocompatibility and wide-spectrum cell proliferation effects on MC3T3-E1 osteoblast cells and human umbilical vein endothelial cells (HUVECs). Moreover, the bridging [SCH2CH2COOH]- thiolate ligands in DNIC-COOH facilitate its chemisorption to deproteinized bovine bone mineral (DBBM) and physisorption onto TCP (ß-tricalcium phosphate), respectively, which provides a mechanism to control the kinetics for the local release of loaded DNIC-COOH. Using rats with calvaria bone defects as an in vivo model, DNIC-DBBM/DNIC-TCP promotes the osteogenic and angiogenic activity ascribed to functional conjugation of osteoconductive bone graft materials and NO-delivery DNIC-COOH. Of importance, the therapeutic efficacy of DNIC-DBBM/DNIC-TCP on enhanced compact bone formation after treatment for 4 and 12 weeks supports the potential for clinical application to regenerative medicine.

Sustained Releasable Copper and Zinc Biogenic Ions Co-Assembled in Metal-Organic Frameworks Reinforced Bacterial Eradication and Wound Mitigation in Diabetic Mice.

The employment of metal-organic framework (MOF)-based nanomaterials has been rapidly increasing in bioapplications owing to their biocompatibility, drug degradation, tunable porosity, and intrinsic biodegradability. This evidence suggests that the multifunctional bimetallic ions can behave as remarkable candidates for infection control and wound healing. In this study, bimetallic MOFs (Zn-HKUST-1 and FolA-Zn-HKUST-1) embedded with and without folic acid were synthesized and used for tissue sealing and repairing incisional wound sites in mice models. For comparison, HKUST-1 and FolA-HKUST-1 were also synthesized. The Brunauer-Emmett-Teller (BET) surface area measured for HKUST-1, FolA-HKUST-1, Zn-HKUST-1, and FolA-Zn-HKUST-1 from N2 isotherms was found to be 1868, 1392, 1706, and 1179 m2/g, respectively. The measurements of contact angle values for Zn-HKUST-1, FolA-HKUST-1, and Zn-FolA-HKUST-1 were identified as 4.95 ± 0.8, 43.6 ± 3.4, and 60.62 ± 2.0°, respectively. For topical application in wound healing, they display a wide range of healing characteristics, including antibacterial and enhanced wound healing rates. In addition, in vitro cell migration and tubulogenic potentials were evaluated. The significant reduction in the wound gap and increased expression levels for CD31, eNOS, VEGF-A, and Ki67 were observed from immunohistological analyses to predict the angiogenesis behavior at the incision wound site. The wound healing rate was analyzed in the excisional dermal wounds of diabetic mice model in vivo. On account of antibacterial potentials and tissue-repairing characteristics of Cu2+ and Zn2+ ions, designing an innovative mixed metal ion-based biomaterial has wide applicability and is expected to modulate the growth of various gradient tissues.

One-step-one-pot hydrothermally derived metal-organic-framework-nanohybrids for integrated point-of-care diagnostics of SARS-CoV-2 viral antigen/pseudovirus utilizing electrochemical biosensor chip.

The COVID-19 pandemic has become a global catastrophe, affecting the health and economy of the human community. It is required to mitigate the impact of pandemics by developing rapid molecular diagnostics for SARS-CoV-2 virus detection. In this context, developing a rapid point-of-care (POC) diagnostic test is a holistic approach to the prevention of COVID-19. In this context, this study aims at presenting a real-time, biosensor chip for improved molecular diagnostics including recombinant SARS-CoV-2 spike glycoprotein and SARS-CoV-2 pseudovirus detection based on one-step-one-pot hydrothermally derived CoFeBDCNH2-CoFe2O4 MOF-nanohybrids. This study was tested on a PalmSens-EmStat Go POC device, showing a limit of detection (LOD) for recombinant SARS-CoV-2 spike glycoprotein of 6.68 fg/mL and 6.20 fg/mL in buffer and 10% serum-containing media, respectively. To validate virus detection in the POC platform, an electrochemical instrument (CHI6116E) was used to perform dose dependent studies under similar experimental conditions to the handheld device. The results obtained from these studies were comparable indicating the capability and high detection electrochemical performance of MOF nanocomposite derived from one-step-one-pot hydrothermal synthesis for SARS-CoV-2 detection for the first time. Further, the performance of the sensor was tested in the presence of Omicron BA.2 and wild-type D614G pseudoviruses.

Fabrication of 3D Amino-Functionalized Metal-Organic Framework on Porous Nickel Foam Skeleton to Combinate Follicle Stimulating Hormone Antibody for Specific Recognition of Follicle-Stimulating Hormone.

In this study, a superficial and highly efficient hydrothermal synthesis method was developed for the in situ growth of amine-functionalized iron containing metal-organic frameworks (H2N-Fe-MIL-101 MOFs) on porous nickel foam (NicF) skeletons (H2N-Fe-MIL-101/NicF). The uniform decoration of the H2N-Fe-MIL-101 nanosheets thus generated on NicF was immobilized with follicle-stimulating hormone (FSH) antibody (Ab-FSH) to detect FSH antigen. In the present work, the Ab-FSH tagged H2N-Fe-MIL-101/NicF electrode was first applied as an immunosensor for the recognition of FSH, electrochemically. With all of the special characteristics, this material demonstrated superior specific recognition and sensitivity for FSH with an estimated detection limit (LOD) of 11.6 and 11.5 fg/mL for buffered and serum solutions, respectively. The availability of specific functional groups on MOFs makes them an interesting choice for exploring molecular sensing applications utilizing Ab-FSH tagged biomolecules.

Utilization of silicon nanowire field-effect transistors for the detection of a cardiac biomarker, cardiac troponin I and their applications involving animal models.

This study develops an ultrasensitive electrical device, the silicon nanowire-field effect transistor (SiNW-FET) for detection of cardiac troponin I (cTnI) in obesity induced myocardial injury. The biosensor device utilizes metal-oxide-semiconductor (MOS) compatible top-down methodology for the fabrication process. After fabrication, the surface of the SiNW is modified with the cTnI monoclonal antibody (Mab-cTnI) upon covalent immobilization to capture cTnI antigen. The sensitivity of the device is also examined using cTnI at different concentrations with the lowest detection limit of 0.016 ng/mL. The electrocardiogram (ECG), magnetic resonance imaging (MRI), and superior vena cave (SVC) provide more information about cardiac responses in a mouse model of acute myocardial infarction (AMI). Further, magnetic resonance imaging helps to evaluate the cardiac output of an obesity induced myocardial injury mouse model. These methods play an essential role in monitoring the obesity based cardiac injury and hence, these studies were carried out. This is the first report to use the ECG, MRI, and SVC sampling methods to study the obesity based cardiac injury involving Syrian hamsters as animal models. The proposed SiNW-FET in this study shows greater sensitivity than the previously developed devices and demonstrates great potential for future applications in point-of-care (POC) diagnosis.

Label-Free Bimetallic In Situ-Grown 3D Nickel-Foam-Supported NH2-MIL-88B(Fe2Co)-MOF-based Impedimetric Immunosensor for the Detection of Cardiac Troponin I.

In this study, a simple and competent metal-organic framework (MOF)-based nickel foam (NF)-supported three-dimensional (3D) immunosensor (Ab-NH2-MIL-88B(Fe2Co)-MOF/NF) was constructed and utilized for the specific recognition of the biomarker cardiac troponin (I) (cTnI). In the present work, biosensor fabrication was progressed through the modification of the NF substrate with the MOF material (NH2-MIL-88B(Fe2Co)-MOF) to enable an amine-functionalized electrode. This amine-functionalized NF electrodes (NH2-MIL-88B(Fe2Co)-MOF/NF) were then biointerfaced with anti-cTnI antibodies, which ended up as Ab-NH2-MIL-88B(Fe2Co)-MOF/NF electrodes. Analytical executions of the constructed bioelectrode were investigated for the quantitative analysis of cTnI in both buffered and serum solutions. Then, the electrochemical studies were carried out using the electrochemical impedance spectroscopy (EIS) method by monitoring changes concerning the charge transfer resistance (Rct) characteristics. The limit of detection (LOD) of the Ab-NH2-MIL-88B(Fe2Co)-MOF/NF immunosensor was achieved to be 13 fg/mL with great specificity. This kind of immunosensor imparts a new platform for the construction and application of MOF-hybrid 3D electrode materials with enhanced electrochemical behavior in cTnI sensing for the first time.

Superparamagnetic iron oxide nanoparticulate system: synthesis, targeting, drug delivery and therapy in cancer.

Cancer is a global epidemic and is considered a leading cause of death. Various cancer treatments such as chemotherapy, surgery, and radiotherapy are available for the cure but those are generally associated with poor long-term survival rates. Consequently, more advanced and selective methods that have better outcomes, fewer side effects, and high efficacies are highly in demand. Among these is the use of superparamagnetic iron oxide nanoparticles (SPIONs) which act as an innovative kit for battling cancer. Low cost, magnetic properties and toxicity properties enable SPIONs to be widely utilized in biomedical applications. For example, magnetite and maghemite (Fe3O4 and γ-Fe2O3) exhibit superparamagnetic properties and are widely used in drug delivery, diagnosis, and therapy. These materials are termed SPIONs when their size is smaller than 20 nm. This review article aims to provide a brief introduction on SPIONs, focusing on their fundamental magnetism and biological applications. The quality and surface chemistry of SPIONs are crucial in biomedical applications; therefore an in-depth survey of synthetic approaches and surface modifications of SPIONs is provided along with their biological applications such as targeting, site-specific drug delivery and therapy.

In Vitro and in Vivo Imaging of Nitroxyl with Copper Fluorescent Probe in Living Cells and Zebrafish.

Nitroxyl (HNO) plays a critical role in many physiological processes which includes vasorelaxation in heart failure, neuroregulation, and myocardial contractility. Powerful imaging tools are required to obtain information for understanding the mechanisms involved in these in vivo processes. In order to develop a rapid and high sensitive probe for HNO detection in living cells and the zebrafish model organism, 2-((2-(benzothiazole-2yl)benzylidene) amino)benzoic acid (AbTCA) as a ligand, and its corresponding copper(II) complex Cu(II)-AbTCA were synthesized. The reaction results of Cu(II)-AbTCA with Angeli's salt showed that Cu(II)-AbTCA could detect HNO quantitatively in a range of 40⁻360 µM with a detection limit of 9.05 µM. Furthermore, Cu(II)-AbTCA is more selective towards HNO over other biological species including thiols, reactive nitrogen, and reactive oxygen species. Importantly, Cu(II)-AbTCA was successfully applied to detect HNO in living cells and zebrafish. The collective data reveals that Cu(II)-AbTCA could be used as a potential probe for HNO detection in living systems.

Ultrasensitive Electrical Detection of Follicle-Stimulating Hormone Using a Functionalized Silicon Nanowire Transistor Chemosensor.

The follicle-stimulating hormone (FSH) is a hormone that belongs to a family of glycoprotein hormones. Determination of FSH can help in interpreting various factors that include physiology of the reproductive system, fertility maintenance, and identification or treatment of reproductive disorders. Sialic acids are derivatives of neuraminic acids with negative charges, present at the end of the sugar chains and further linked to the cell surfaces and glycoproteins. The direct measurement of FSH in a human body can be recorded by developing a sensor probe that responds particularly to sialic acids over the other hormones. However, existing diagnostic methods still suffer from many difficulties in terms of complicated handling techniques, expensive instrumentation, etc. Development of accurate, rapid, and low-cost FSH detection chemosensors is important to meet these demands. Herein, we utilized a novel sensing method for accurate and fast FSH detection using a metal-oxide semiconductor silicon nanowire field effect transistor (SiNW-FET) device. This is the first report to demonstrate the boronic acid-functionalized SiNW-FET device in FSH detection. FSH detection has been successfully determined using an assay buffer solution with 0.72 fM detection limit as well as using 20% serum with 1.1. fM detection limit. We also investigated the specificity with other gonadotropins/glycosylated serum proteins. The current measurements on FSH concentrations at different time intervals were also studied. The sensitive, cheap, and miniaturized SiNW-FET device can serve as an effective sensing approach for rapid screening of FSH and menopause diagnosis.

In vitro and in vivo imaging of peroxynitrite by a ratiometric boronate-based fluorescent probe.

Peroxynitrite (ONOO-) is an important species involved in many physiopathological processes. Progresses have been made in developing novel fluorescent probes to detect peroxynitrite with relatively high sensitivity and specificity. Herein, we report the synthesis, characterization and biological applications of a new boronate-based fluorescent probe, 4-MB. The studies showed that 4-MB exhibits a dual ratiometric and calorimetric response toward peroxynitrite due to ONOO--triggered oxidative reaction. A possible mechanism of the oxidation reaction was proposed and the reaction product was isolated and characterized using different spectroscopic methods. We have thoroughly demonstrated the utility of 4-MB for intracellular peroxynitrite imaging. Further, we showed that 4-MB can be potentially employed to visualize exogenous and endogenous peroxynitrite in RAW264.7 macrophages, EAhy926 cells, zebrafish and in live tissues from a high-fat diet-induced obese mouse model.

A water soluble and fast response fluorescent turn-on copper complex probe for H2S detection in zebra fish.

According to the displacement method, herein we reported a water soluble copper complex [Cu(MaT-cyclen)2] as a fluorescent probe for the detection of H2S. For this, 1-((1-((10-methylanthracen-9-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)-1,4,7,10-tetraazacyclododecane (MaT-cyclen) was synthesized first. To improve its solubility in aqueous media, sodium acetate group was introduced into 8-hydroxy-2-quinoline successfully. MaT-cyclen was chelated with Cu(II) to form [Cu(MaT-cyclen)2] complex, which displayed high sensitivity and selectivity for H2S over the other possible competitive substances on the basis of forming CuS. Meanwhile, [Cu(MaT-cyclen)2] displayed rapid response (<1min), well reversibility, lowest detection limit (205nM), and high sensitivity for recognizing H2S in aqueous solution. Furthermore, its potential utility for biological applications was confirmed by fluorescence imaging of H2S in live cells as well as in zebra fish.