Synthesis, structural characterization, and biological studies of ATBS-M complexes (M(II) = Cu, Co, Ni, and Mn): Access for promising antibiotics and anticancer agents.

A new bidentate Schiff base ligand (ATBS [4-bromo-2-(thiazole-2-yliminomethyl)phenol]) was synthesized via the condensation reaction of 2-aminothiazole with 5-bromosalicylaldehyde in ethanol. The reaction of ATBS with transition metal salts of Cu(II), Co(II), Ni(II), and Mn(II) afforded the corresponding ATBS-M complexes. Results from physicochemical and spectral analyses, such as elemental analysis, infrared, UV-Vis spectroscopy, magnetic susceptibility, and molar conductance, revealed a nonelectrolytic nature with octahedral (Oh ) geometry and a metal/ligand ratio of 1:2 for Cu(II), Co(II), and Ni(II), but 1:1 for the Mn(II) complex. The density functional theory (DFT) calculations are correlated very well with the proposed structure and molecular geometry of the complexes as [M(ATBS)2 ] (M = Cu, Co, and Ni) and [Mn(ATBS)(H2 O)2 ]. Significantly, the prepared compounds showed strong inhibition activity for a wide spectrum of bacteria (Escherichia coli, Bacillus subtilis, and Staphylococcus aureus) and fungi (Candida albicans, Aspergillus flavus, and Trichophyton rubrum), with the ATBS-Ni complex being the most promising antibiotic agent. Molecular docking studies of the binding interaction between the title complexes with the bacterial protein receptor CYP51 revealed clear insights about the inhibition nature against the studied microorganisms, with the following order: ATBS-Cu > ATBS-Mn > ATBS-Ni > ATBS-Co for complex stability. Moreover, the cytotoxicity measurements of all prepared metal complexes against the colon carcinoma (HCT-116) and hepatocellular carcinoma (Hep-G2) cell lines showed exceptional anticancer efficacy of the complexes as compared with the free ATBS Schiff base ligand. Significantly, the results attested that ATBS-Cu is the most effective complex against HCT-116 cells, whereas ATBS-Mn has the highest cytotoxic efficiency against Hep-G2 cells. Furthermore, electronic spectra, viscosity measurements, and gel electrophoresis techniques were employed to probe the interaction of all prepared ATBS-metal complexes with calf thymus (CT)-DNA. Results confirmed that all complexes are strongly bound to CT-DNA via intercalation mode, with the ATBS-Co complex having the highest binding ability.

Thiacloprid impairs reproductive functions of male Wistar rats.

Global male infertility correlated to the rise of endocrine-disrupting chemicals, including insecticides, has grown into a pressing problem. Thiacloprid is one of the most commonly used neonicotinoids that accounts for more than 25% of the global pesticide industry. However, its impact on the reproductive system and male fertility has not been fully elucidated. The object of this study was to explore the adverse effects of thiacloprid on male Wistar rats' reproductive system. Thirty healthy male rats were separated into one of three groups: control group, and two groups that were orally administered with low (22.5 mg/kg) and high dose (62.1 mg/kg) of thiacloprid for 56 days. Thiacloprid significantly (p<0.05) reduced body weight and relative testicular weight, as well as sperm quality (count, motility, viability, and morphology), in a dose-dependent manner. THIA-treated groups revealed a large effect (d > 0.8) on semen quality with Cohen's d of (6.57, 8.82), (20.14, 23.54), and (2.81, 9.10) for count, motility, and viability respectively. Meanwhile, the serum testosterone level dropped while the levels of luteinizing and follicle-stimulating hormones increased. 17ꞵ-hydroxy steroid dehydrogenase and 3ꞵ-hydroxy steroid dehydrogenase levels were significantly decreased in a dose-dependent manner. The activity of the tested antioxidant enzymes catalase (CAT), glutathione reduced (GSH), and superoxide dismutase (SOD) exhibited a considerable decrease compared to the control group with a significant elevation in the lipid peroxidation activity as indicated by malondialdehyde (MDA) level. The testicular histology revealed degenerative changes in spermatogenic cells and interstitial tissue. Comet assay revealed DNA fragmentation in treated groups' testicular tissue. Thiacloprid exposure interferes with reproductive function and impairs male Wistar rat fertility. Such harmful consequences may also develop in humans frequently exposed to thiacloprid.

X-Ray Visible Protein Scaffolds by Bulk Iodination.

Protein-based biomaterial use is expanding within medicine, together with the demand to visualize their placement and behavior in vivo. However, current medical imaging techniques struggle to differentiate between protein-based implants and surrounding tissue. Here a fast, simple, and translational solution for tracking transplanted protein-based scaffolds is presented using X-ray CT-facilitating long-term, non-invasive, and high-resolution imaging. X-ray visible scaffolds are engineered by selectively iodinating tyrosine residues under mild conditions using readily available reagents. To illustrate translatability, a clinically approved hernia repair mesh (based on decellularized porcine dermis) is labeled, preserving morphological and mechanical properties. In a mouse model of mesh implantation, implants retain marked X-ray contrast up to 3 months, together with an unchanged degradation rate and inflammatory response. The technique's compatibility is demonstrated with a range of therapeutically relevant protein formats including bovine, porcine, and jellyfish collagen, as well as silk sutures, enabling a wide range of surgical and regenerative medicine uses. This solution tackles the challenge of visualizing implanted protein-based biomaterials, which conventional imaging methods fail to differentiate from endogenous tissue. This will address previously unanswered questions regarding the accuracy of implantation, degradation rate, migration, and structural integrity, thereby accelerating optimization and safe translation of therapeutic biomaterials.

Recellularization of Native Tissue Derived Acellular Scaffolds with Mesenchymal Stem Cells.

The functionalization of decellularized scaffolds is still challenging because of the recellularization-related limitations, including the finding of the most optimal kind of cell(s) and the best way to control their distribution within the scaffolds to generate native mimicking tissues. That is why researchers have been encouraged to study stem cells, in particular, mesenchymal stem cells (MSCs), as alternative cells to repopulate and functionalize the scaffolds properly. MSCs could be obtained from various sources and have therapeutic effects on a wide range of inflammatory/degenerative diseases. Therefore, in this mini-review, we will discuss the benefits using of MSCs for recellularization, the factors affecting their efficiency, and the drawbacks that may need to be overcome to generate bioengineered transplantable organs.

Decellularized extracellular matrix-rich hydrogel-silver nanoparticle mixture as a potential treatment for acute liver failure model.

Acute liver failure (ALF) occurs due to severe liver damage that triggers rapid loss of normal liver function. Here, we investigate the usefulness of an injectable liver extracellular matrix (LECM)-rich hydrogel generated from an optimized decellularization protocol incorporated with silver nanoparticles (AgNPs) as a promising therapy for ALF. First, we optimized a non-destructive protocol for rat liver decellularization to obtain ECM-rich well-preserved scaffold. Then, LECM hydrogel generated from two commonly used decellularization protocols were compared by LECM hydrogel obtained from our optimized protocol. The ALF model was induced by an intraperitoneal (IP) thioacetamide (TAA) injection followed by the IP injection of LECM hydrogel, collagen-AgNP mixture, or LECM hydrogel-AgNP mixture. LECM-rich scaffold and hydrogel were successfully obtained using our optimized decellularization protocol. Use of the LECM hydrogel-AgNP mixture to treat TAA-induced ALF greatly improved liver injury and histological liver regeneration. Interleukin-6 and transforming growth factor-beta expressions were significantly reduced, while albumin, hepatocyte growth factor, and Ki67-positive cells were highly expressed. Moreover, aspartate transaminase and alanine transaminase plasma levels and liver homogenate nitric oxide level were significantly lowered. In conclusion, the LECM hydrogel-AgNP mixture has potential efficient therapeutic and regenerative effects on TAA-induced liver injury.

Conjugating homogenized liver-extracellular matrix into decellularized hepatic scaffold for liver tissue engineering.

The generation of a transplantable liver scaffold is crucial for the treatment of end-stage liver failure. Unfortunately, decellularized liver scaffolds suffer from lack of bioactive molecules and functionality. In this study, we conjugated homogenized liver-extracellular matrix (ECM) into a decellularized liver in a rat model to improve its structural and functional properties. The homogenized ECM was prepared, characterized, and subsequently perfused into ethyl carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) activated liver scaffolds. Various techniques were performed to confirm the improvements that were accomplished through the conjugation process; these included micro/ultra-structural analyses, biochemical analysis of ECM components, DNA quantification, swelling ratio, structural stability, calcification properties, platelet activation study, static and dynamic seeding with EAhy926 endothelial cells and HepG2 hepatocarcinoma cells, subcutaneous implantation and intrahepatic transplantation. The results showed that the conjugated scaffolds have superior micro- and ultrastructural and biochemical characteristics. In addition, DNA contents, swelling ratios, calcification properties, platelet reactions, and host inflammatory reactions were not altered with the conjugation process. The conjugated scaffolds revealed better cellular spreading and popularity compared to the non-conjugated scaffolds. Intrahepatic transplantation showed that the conjugated scaffold had higher popularity of hepatic regenerative cells with better angiogenesis. The conjugation of the decellularized liver scaffold with homogenized liver-ECM is a promising tool to improve the quality of the generated scaffold for further transplantation.

Micro and ultrastructural changes monitoring during decellularization for the generation of a biocompatible liver.

Decellularization of a whole organ is an attractive process that has been used to create 3D scaffolds structurally and micro-architecturally similar to the native one. Currently used decellularization protocols exhibit disrupted extracellular matrix (ECM) structure and denatured ECM proteins. Therefore, maintaining a balance between ECM preservation and cellular removal is a major challenge. The aim of this study was to optimize a multistep Triton X-100 based protocol (either using Triton X-100/ammonium hydroxide mixture alone or after its modification with DNase, sodium dodecyl sulfate or trypsin) that could achieve maximum decellularization with minimal liver ECM destruction suitable for subsequent organ implantation without immune rejection. Based on our findings, Triton X-100 multistep protocol was insufficient for whole liver decellularization and needed to be modified with other detergents. Among all Triton X-100 modified protocols, a Triton X-100/DNase-based one was considered the most suitable. It maintains a gradual but sufficient removal of cells to generate decellularized biocompatible liver scaffolds without any significant alteration to ECM micro- and ultra-structure.

Characterization of silver nanoparticle-modified decellularized rat esophagus for esophageal tissue engineering: Structural properties and biocompatibility.

Decellularized esophageal matrices are ideal scaffolds for esophageal tissue engineering. Unfortunately, in order to improve transplantation possibilities, they require modification to reduce their degradation rate and immunogenicity. To date, no modifying agent has been approved to overcome these limitations. The objective of this study was to evaluate the ability of silver nanoparticles (AgNPs) to improve the structural stability and biocompatibility of decellularized rat esophagi. AgNPs have the advantage over currently used agents in that they bind with collagen fibers in a highly ordered manner, via non-covalent binding mechanisms forming multiple binding sites, while other agents provide only two-point connections between collagen molecules. Rat esophagi were decellularized, loaded with 5 µg/mL of AgNPs (100 nm), and then treated with an immobilization-complex buffer composed of ethyl carbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS). Then, they were evaluated in terms of ultra-structural morphology, water uptake, in vitro resistance to enzymatic and thermal degradation, indentation strength, in vitro anti-calcification, cytocompatibility with rat bone marrow derived stromal cells (rat-BMSCs), angiogenic properties, and in vivo biocompatibility, and compared to scaffolds modified using glutaraldehyde and EDC/NHS complex buffer alone. AgNP-modified scaffolds showed an improved ultrastructure, good water uptake, and considerable resistance against in vitro degradation and indentation, and a high resistance against in vitro calcification. Moreover, they were cytocompatible for allogeneic rat-BMSCs. Additionally, AgNPs did not alter the angiogenic properties of the modified scaffolds and decreased host immune responses after their subcutaneous implantation. The structural properties and biocompatibility of decellularized esophageal matrices could be improved by conjugation with AgNPs.

Biocompatibility and hemocompatibility of efficiently decellularized whole porcine kidney for tissue engineering.

Whole kidney decellularization is a promising approach in regenerative medicine for engineering a functional organ. The reaction of the potential host depends on the biocompatibility of these decellularized constructs. Despite the proven ability of decellularized kidney scaffolds to guide cell attachment and growth, little is known about biocompatibility and hemocompatibility of these scaffolds. Our aim is to prepare decellularized kidneys of a clinically relevant size and evaluate its biocompatibility and hemocompatibility. Porcine kidneys were cannulated via the renal artery, and then perfused with 0.1% sodium dodecyl sulfate solution. Hematoxylin and eosin as well as DAPI staining confirmed cellular clearance from native kidneys in addition to preservation of the microstructure. SEM confirmed the absence of any cellular content within the scaffold, which is maintained in a well-organized 3D architecture. Decellularized kidneys retained the intact renal vasculature upon examination with contrast radiography. The essential structural extracellular matrix molecules were well-preserved. Scaffolds were susceptible to enzymatic degradation upon collagenase treatment. Scaffolds showed a good hemocompatibility when exposed to porcine blood. Decellularization was efficient to remove 97.7% of DNA from native kidneys in addition to the immunogenic and pathogenic antigens. Scaffolds did not induce the human immune response in vitro. Decellularized kidneys were non-cytotoxic to pig kidney cells (PKs). PKs were able to grow and proliferate within the decellularized renal scaffolds with maintaining a higher function than cells grown as monolayers. Thus, we have developed a rapid decellularization technique for generating biocompatible kidney scaffolds that represents a step toward development of a transplantable organ. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2034-2047, 2018.

Incorporation of nanoparticles into transplantable decellularized matrices: Applications and challenges.

Decellularization of tissues can significantly improve regenerative medicine and tissue engineering by producing natural, less immunogenic, three-dimensional, acellular matrices with high biological activity for transplantation. Decellularized matrices retain specific critical components of native tissues such as stem cell niche, various growth factors, and the ability to regenerate in vivo. However, recellularization and functionalization of these matrices remain limited, highlighting the need to improve the characteristics of decellularized matrices. Incorporating nanoparticles into decellularized tissues can overcome these limitations because nanoparticles possess unique properties such as multifunctionality and can modify the surface of decellularized matrices with additional growth factors, which can be loaded onto the nanoparticles. Therefore, in this minireview, we highlight the various approaches used to improve decellularized matrices with incorporation of nanoparticles and the challenges present in these applications.

Silver nanoparticles improve structural stability and biocompatibility of decellularized porcine liver.

No ideal cross-linking agent has been identified for decellularized livers (DLs) yet. In this study, we evaluated structural improvements and biocompatibility of porcine DLs after cross-linking with silver nanoparticles (AgNPs). Porcine liver slices were decellularized and then loaded with AgNPs (100 nm) after optimization of the highest non-toxic concentration (5 µg/mL) using Human hepatocellular carcinoma (HepG2) and EAhy926 human endothelial cell lines. The cross-linking effect of AgNPs was evaluated and compared to that of glutaraldehyde and ethyl carbodiimide hydrochloride and N-hydroxysuccinimide. The results indicated that AgNPs improved the ultra-structure of DLs' collagen fibres with good porosity and increased DLs' resistance against in vitro degradation with good cytocompatibility. AgNPs decreased the host inflammatory reaction against implanted porcine DL slices in vivo and increased the polarization of M2 macrophages. Thus, structural and functional improvements of Porcine DLs could be achieved using AgNPs.