Bovine Serum Research Articles

Automated Manufacturing Processes and Platforms for Large-scale Production of Clinical-grade Mesenchymal Stem/ Stromal Cells.

Mesenchymal stem/stromal cells (MSCs) hold immense potential for regenerative medicine due to their remarkable regenerative and immunomodulatory properties. However, their therapeutic application requires large-scale production under stringent regulatory standards and Good Manufacturing Practice (GMP) guidelines, presenting significant challenges. This review comprehensively evaluates automated manufacturing processes and platforms for the scalable production of clinical-grade MSCs. Various large-scale culture vessels, including multilayer flasks and bioreactors, are analyzed for their efficacy in MSCs expansion. Furthermore, automated MSCs production platforms, such as Quantum® Cell Expansion System, CliniMACS Prodigy®, NANT001/ XL, CellQualia™, Cocoon® Platform, and Xuri™ Cell Expansion System W25 are reviewed and compared as well. We also underscore the importance of optimizing culture media specifically emphasizing the shift from fetal bovine serum to humanized or serum-free alternatives to meet GMP standards. Moreover, advances in alternative cryopreservation methods and controlled-rate freezing systems, that offer promising improvements in MSCs preservation, are discussed as well. In conclusion, advancing automated manufacturing processes and platforms is essential for realizing the full potential of MSCs-based regenerative medicine and accomplishing the increasing demand for cell-based therapies. Collaborative initiatives involving industry, academia, and regulatory bodies are emphasized to accelerate the translation of MSCs-based therapies into clinical practice.

On the Non-Catalytic Role of Lytic Polysaccharide Monooxygenases in Boosting the Action of PETases on PET Polymers.

Synthetic polymers are resistant to biological attack, resulting in their long-term accumulation in landfills and in natural aquatic and terrestrial habitats. Lytic polysaccharide monooxygenases (LPMOs) are enzymes which oxidatively cleave the polysaccharide chains in recalcitrant polysaccharides such as cellulose. It has been widely hypothesised that LPMOs could be used to aid in the enzymatic breakdown of synthetic polymers. Herein, through the use of biochemical assays, X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) we show that LPMOs can bind to polyethylene terephthalate (PET), and - in doing so - the hydrophobic surface of PET becomes more hydrophilic such that product release is boosted by subsequent treatment with classical PETases. The boosting effect is however, only observed in reactions when the LPMO and the PETase are added sequentially rather than simultaneously to the PET. Moreover, the same boosting effect is also seen when a catalytically-inactive mutant of LPMO is used, showing that the principal means by which AA9 LPMOs boost the degradation of synthetic polymers is through their role as a "hydrophobin" rather than as an oxygenase. Indeed, in accord with this role of LPMOs, we further show that this effect can be extended to other ostensibly 'non-catalytic' proteins beyond LPMOs, such as bovine serum albumin and lactate dehydrogenase.

Virus susceptibility of a new cell line derived from the muscle of koi (Cyprinus carpio koi).

In this study, a continuous cell line (KM cells) derived from koi (Cyprinus carpio koi) muscle was established and characterized. The KM cells were subcultured for more than 70 passages and showed high viability after long-term cryopreservation. The KM cell line was optimally cultured in medium 199 containing 10% foetal bovine serum at 25°C. A chromosome analysis indicated that the cell line remained diploid, with a mean chromosome count of 100. DNA sequencing and comparative analysis of the 16S rRNA and cytochrome oxidase I gene sequences showed that the KM cell line originated from koi. In transfection experiments using the plasmid pEGFP, KM cells demonstrated a high level of transfection efficiency, suggesting their potential for use in foreign gene expression studies. Inoculation with spring viraemia of carp virus (SVCV) resulted in a substantial cytopathic effect, and the level of production of SVCV in KM cells was higher than that in the epithelioma papulosum cyprinid (EPC) cell line that is normally used to produce the virus. However, no cytopathic effect was observed when these cells were inoculated with koi herpesvirus, carp oedema virus, or grass carp reovirus. These observations suggest that the newly established KM cell line will be a valuable tool for investigating the pathogenesis of infection with spring viraemia of carp virus.

Exactly defined molecular weight poly(ethylene glycol) allows for facile identification of PEGylation sites on proteins.

PEGylation (the covalent attachment of one or more poly(ethylene glycol) (PEG) units to a therapeutic) is a well-established technique in the pharmaceutical industry to increase blood-residence time and decrease immunogenicity. A challenging aspect of PEGylation is the dispersity of PEGylation agents, which results in batch-to-batch variations and analytical limitations. Herein, we present an approach to overcome these limitations by manufacturing a defined molecular weight (dispersity-free) PEGylation agent. We synthesise a defined molecular weight (Mw), linear 5 kDa methoxy-PEG (mPEG) active ester in an efficient and scalable manner using an iterative liquid-phase approach based on Nanostar Sieving. We then perform a comparative study on the random PEGylation and subsequent characterisation of the protein bovine serum albumin (BSA), using both the defined Mw, dispersity-free mPEG active ester, and a commercially available disperse 5 kDa mPEG active ester. We demonstrate that the defined Mw PEG both allows for facile monitoring of chemical modification reactions during the synthesis of the PEGylation agents, and facilitates straightforward identification of the PEGylated fragments within a PEGylated protein via a simple peptide mapping approach using UPLC-MS.

Membranes fouling propensity of PSF/GO hollow fiber mixed matrix membranes for water treatment ultrafiltration application.

The study investigated the fouling propensity of polysulfone (PSF) hollow fiber (HF) mixed matrix membranes modified with 1.0 wt.% graphene oxide (GO). Using scanning electron microscopy (SEM), atomic force microscopy (AFM), water contact angle (WCA), and mechanical assessments, the structural characteristics of both untreated and GO-modified PSF HF membranes were examined. Filtration experiments included pure water and model contaminants such as bovine serum albumin (BSA), humic acid (HA), E. coli, and oil-in-water emulsion. The GO-modified membranes demonstrated a significant enhancement in antifouling performance, recovering over 90% of their initial pure water flux with HA and oil, indicating high resistance to irreversible fouling. Additionally, the GO-modified membranes showed superior oil separation efficiency. However, fouling parameters for BSA were similar for both membrane types, suggesting that GO does not significantly affect membrane-BSA interactions. Both types of membranes displayed high retention capabilities for E. coli, with no noticeable improvement due to GO addition. This study highlights the potential of GO-modified PSF HF membranes in enhancing antifouling performance and oil separation efficiency.

A Multi-Functional Cascade Nanoreactor for Remodeling Tumor Microenvironment to Realize Mitochondria Dysfunction via ROS/Zn2+ Ions Overload.

Combining chemo/photodynamic therapy (CDT/PDT) to generate highly harmful reactive oxygen species and cause mitochondria dysfunction is considered a potential strategy to improve the efficiency of anticancer treatment. However, within tumor, the relatively deficient concentration of H2O2, hypoxic microenvironment, and overexpressed reduced glutathione (GSH)seriously suppress the efficacy of dynamic therapy. Herein, a multi-functional cascade nanoreactor, bovine serum albumin modified ZnO2@CeO2-ICG, is reported for remodeling tumor microenvironment (TME) to boost dynamic therapy and realize mitochondria dysfunction via reactive oxygen species (ROS) storm/Zn2+ ions overload. Within TME, ZnO2 decomposed into exogenous H2O2 and Zn2+ ion. The dual enzyme-like CeO2 catalyzes the increased H2O2 into ·OH and oxygen molecules respectively, and then the oxygen molecules are translated into 1O2 by indocyanine green (ICG) under 808 nm light irradiation to boost PDT. The effective consumption of GSH through the reduction of Ce(IV) ions not only regenerates Ce(III) ions to enhance the efficiency of CDT but also efficaciously alleviates the elimination of ROS generated by dynamic therapy to further improve dynamic therapeutic efficiency. So the improved ROS level under remodeling TME and Zn2+ ions acutely lead to mitochondria dysfunction to boost the efficiency of antitumor treatment. Thus, developing functional nanoreactors that enable remodeling TME provides a potential strategy to enhance the efficiency of dynamic therapy.

Translational implications of a novel combination of iPRF and collagen scaffold for proliferation of gingival mesenchymal stem cells.

"Tissue engineering," is a concept that involves the use of scaffolds, cells, and growth factors/signaling molecules in the field of regenerative medicine. The present study was carried out to evaluate the attachment and depth of penetration of the Gingival Mesenchymal Stem Cells (GMSCs) onto the collagen scaffold preconditioned with Fetal Bovine Serum (FBS), Injectable Platelet Rich Fibrin (i-PRF) and a combination of FBS with i-PRF using histological analysis and environmental scanning electron microscopy (ESEM) respectively. In the present study, commercially available collagen membranes were used as scaffolds and divided into 3 groups where Group I was preconditioned with FBS, Group II was preconditioned with i-PRF, and Group III with a combination of FBS and i-PRF. Scaffolds were then seeded with GMSCs and incubated in a CO2 incubator at 37 0C for 7 days. Both histological and ESEM analysis showed proliferation, attachment, and depth of penetration along with the combination of cell morphological changes in all the three groups. In group I cells showed mild depth of penetration along with a round morphological appearance. In group II the cells showed moderate depth of penetration and rounded morphology. In group III maximum depth of penetration was seen along with the round and spheroidal morphology of cells. The combination of the growth media showed the best effect on cell attachment, depth of penetration, and cell morphology. This is the first report demonstrating the effect of the combination of collagen and i-PRF supports the proliferation of GMSCs. Since FBS alone and i-PRF alone exhibited similar cell proliferation it is recommended that i-PRF could be used in clinical scenarios making it xenofree.

PH and Redox Dual-Responsive Nanoparticle with Enhanced Dendritic Cell Maturation for Cancer Therapy.

Type I interferons (IFNs) are essential for activating dendritic cells (DCs) and presenting tumor-associated antigens to T cells. IFNs are primarily produced from DCs among immune cells. A combination of chemotherapy and metalloimmunotherapy induces IFN production by activating the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway. However, chemotherapeutic agents deplete DC populations, suppressing immunostimulatory activities, despite their potent anticancer activities. Furthermore, an optimal ratio between chemotherapeutic agents and metal for activating DCs at the highest level has not been reported, and evidence for ensuring DC survival is lacking. In this study, we hypothesized that there is an optimal ratio to yield the highest DC maturation and anticancer activity with minimal DC depletion. To demonstrate it, we have designed a pH and redox dual-responsive nanoparticle, MnO2@BSA@DOX (MD), to prevent DCs from depleting and activate the cGAS-STING pathway both in cancer cells and DCs, inducing considerable levels of IFNs and maturation. MD consists of a core-layer structure, a manganese dioxide (MnO2) core, and a cross-linked layer with bovine serum albumin (BSA) and doxorubicin (DOX), with a specific ratio of DOX to manganese. MD exhibits structure-based selectivity between cancer cells and DCs by targeting the extracellular pH of the tumor microenvironment and intracellular redox reactions in cancer cells. Among various formulations, the 1:1 ratio shows the highest maturation with no significant depletion. Moreover, it induces distinct cytotoxicity in cancer cells through apoptosis and cGAS-STING activation, leading to increased calreticulin expression and enhanced DC phagocytosis. Consequently, it results in superior tumor suppression and prolonged survival with the high accumulation of MD in the tumor and no observed systemic toxicities, highlighting its potential as a therapeutic agent in cancer treatments.

Nanotherapeutics-mediated restoration of pancreatic homeostasis and intestinal barrier for the treatment of severe acute pancreatitis.

Severe acute pancreatitis (SAP) is an inflammatory disease of the pancreas accompanied with intestinal injury, and effective therapeutic modalities are still highly lacking. Herein, a facile and effective nanotherapeutics (pHA@IBNCs) is developed to alleviate pancreatic inflammation and restore intestinal barrier for SAP treatment. Epigallocatechin gallate (EGCG, an anti-oxidant), interleukin-22 (IL-22, an anti-inflammatory and epithelial barrier-protecting cytokine), and bovine serum albumin (a framework protein), are assembled via non-covalent interactions to form nanocomplexes (IBNCs). Then, phenylboronic acid-modified hyaluronic acid (pHA) is synthesized and coated onto IBNCs via formation of the reversible boronate ester bonds to obtain pHA@IBNCs. Upon intravenous injection, pHA@IBNCs could efficiently accumulate at the lesion sites of sodium taurocholate (STC)-induced SAP mice, based on their prolonged blood circulation time and pHA-mediated targeting of activated intestinal epithelial cells and macrophages. Inside the inflammatory microenvironment, over-produced reactive oxygen species (ROS) trigger the shedding of the pHA layer and release of the drug payloads. Thereby, EGCG cooperates with IL-22 to attenuate pancreatitis and restore the intestinal barrier by scavenging ROS, suppressing pro-inflammatory cytokines secretion, and promoting the repair of intestinal epithelia. Such a nano-therapeutic approach targeting multiple pathological events may serve as a promising paradigm for the effective management of SAP.

PH-Responsive Modified HAMA Microspheres Regulate the Inflammatory Microenvironment of Intervertebral Discs.

Currently, intervertebral disc (IVD) degeneration is believed to lead to local accumulation of lactic acid in the IVD, a decrease in pH, activation of the inflammatory pathway, and continued destruction of homeostasis of the IVD. To address these issues, the intelligent and accurate release of drugs is particularly important. In this study, acid-sensitive release methacrylated hyaluronic acid (HAMA) microspheres were constructed by using microfluidic technology, which can be used as a targeted drug delivery system for intervertebral disc degeneration (IVDD) through Schiff base chemical bonding on the surface of the microspheres to achieve intelligent drug release. Interleukin-1 receptor antagonist (IL-1 Ra) is a naturally occurring anti-inflammatory antagonist of the interleukin-1 family of pro-inflammatory cytokines. Despite its outstanding broad-spectrum anti-inflammatory effects, IL-1 Ra has a short biological half-life (4-6 h). The slow-release performance of IL-1 Ra can be greatly improved using bovine serum albumin nanoparticles (BNP). In addition, the modified HAMA microspheres exhibited good injectability and porosity, and efficient and uniform loading of nanoparticles was achieved via the Schiff base bond. The inflammatory microenvironment can be significantly reversed by transporting the modified HAMA microspheres-BNPs (Modified MS) to the degenerative nucleus pulposus.

Impact of coverage and guest residue on polyproline II helix peptide antifouling

AbstractPolyproline II (PPII) peptide sequences are recognized as promising biomaterials because of their attractive antifouling properties. However, the mechanisms behind their antifouling behavior have not been fully characterized. In this work we show that PPII peptide coverage, controlled by adsorption time, significantly reduces the fouling of bovine serum albumin (BSA, a model foulant). In addition, guest residues introduced into the PPII sequence are shown to significantly impact BSA adsorption as well as human mesenchymal stem cell (hMSC) spreading. This research will help guide future PPII peptide designs for incorporation into novel biomaterials. Graphical abstract

Albumin-Chaperoned Deep-NIR Triarylmethane Dyes for High-Contrast In Vivo Imaging and Photothermal Therapy.

Fluorophores absorbing/emitting in the deep near-infrared (deep NIR) spectral region, that is, 800 nm and beyond, hold great promise for in vivo bioimaging, diagnosis, and phototherapy due to deeper tissue penetration. The bottleneck is the lack of bright, stable, and readily synthesized deep NIR fluorophores. Here, it is reported that the albumin-chaperon strategy is a viable one-for-all strategy to address these difficulties. A focused library of deep-NIR absorbing dyes (EA5) is easily synthesized via a two-step cascade. They are neither very stable nor bright in phosphate bufferdue to a propeller-type flexible scaffold. Through screening, EA5_c3 is found to exhibit a high affinity toward bovine serum albumin (BSA). Binding-associated structural rigidification resulted in a gigantic 26-fold fluorescence enhancement. The albumin chaperone also greatly improved the stability of EA5_c3 by shielding the bisbenzannulated triarylmethane core from nucleophilic or oxidative species. The resulting EA5_c3@BSA exhibits high biocompatibility. It offered high-resolution vasculature, lymph systems, tumors, and other tissue imaging with its bright deep NIR emission. At the same time, it exhibits prominent potential in photoacoustic imaging and photothermal treatment of subcutaneous and orthotopic breast tumors. These findings provide insights into robust and high-performance fluorophores with deep NIR regions for theranostic against aggressive cancers.

Combined Effects of Surface Roughness, Solubility Parameters, and Hydrophilicity on Biofouling of Reverse Osmosis Membranes

The fouling of protein on the surface of reverse osmosis (RO) membranes is a surface phenomenon strongly dependent on the physical and chemical characteristics of both the membrane surface and the foulant molecule. Much of the focus on fouling mitigation is on the synthesis of more hydrophilic membrane materials. However, hydrophilicity is only one of several factors affecting foulant attachment. A more systematic and rationalized methodology is needed to screen the membrane materials for the synthesis of fouling-resistant materials, which will ensure the prevention of the accumulation of foulants on the membrane surfaces, avoiding the trial and error methodology used in most membrane synthesis in the literature. If a clear correlation is found between various membrane surface properties, in combination or singly, and the amount of fouling, this will facilitate the establishment of a systematic strategy of screening materials and enhance the selection of membrane materials and therefore will reflect on the efficiency of the membrane process. In this work, eight commercial reverse osmosis membranes were tested for bovine serum albumin (BSA) protein fouling. The work here focused on three surface membrane properties: the surface roughness, the water contact angle (hydrophilicity), and finally the Hansen solubility parameter (HSP) distance between the foulant understudy (BSA protein) and the membrane surface. The HSP distance was investigated as it represented the affinities of materials to each other, and therefore, it was believed to have an important contribution to the tendency of foulant to stick to the surface of the membrane. The results showed that the surface roughness and the HSP distance contributed to membrane fouling more than the hydrophilicity. We recommend taking into account the HSP distance between the membrane material and foulants when selecting membrane materials.

Polyethersulfone mixed matrix membranes modified with pore formers and Ag-titanate nanotubes: physicochemical characteristics and (bio)fouling study.

In the presented studies it was hypothesized that the modification of a polymeric membrane with a pore former and a hybrid nanomaterial composed of titanate nanotubes with deposited Ag nanoparticles (Ag-TNTs NPs) can protect the membrane from the microbial growth, and thus enhance its resistance to biofouling. Polyethersulfone (PES) membranes were prepared by the wet phase inversion, and polyvinylpyrrolidone (PVP) and poly(ethylene glycol) (PEG) were used as pore formers. The membranes were characterized in terms of morphology, topography, permeability, separation characteristics, and anti-(bio)fouling properties as well as antibacterial activity. The membranes modified with porogens and Ag-TNTs revealed improved hydrophilicity and water permeability compared to the unmodified membrane, from 58 to 66%. Moreover, the improvement in rejection of model dextrans and PEG upon application of the NPs was found. However, the use of PVP or PEG had a negative influence on the resistance to fouling by bovine serum albumin, i.e., ca. 35% of decline of permeate flux was noticed after 2h of ultrafiltration of BSA. On the contrary, both porogens and NPs contributed to biofouling mitigation. The introduction of pore formers had a positive effect on the inhibition of Escherichia coli growth by the membrane containing Ag-TNTs. The log reduction of bacteria varied from 3.17 to 3.3 in case of stirred and filtration system.

Porphyrin-Polymer as a Photosensitizer Prodrug for Antimicrobial Photodynamic Therapy and Biomolecule Binding Ability.

This study presents the development and characterization of a novel porphyrin-Jeffamine polymer conjugate designed to function as a photosensitizer prodrug for antimicrobial photodynamic therapy (aPDT). The conjugate features a photosensitive porphyrin unit covalently attached to a biocompatible polymer backbone, with enhanced solubility, stability, and bioavailability compared to those of the free porphyrin derivatives. The photophysical properties were studied using transient absorption spectroscopy spanning the fs-μs time scales in addition to emission studies. The production of reactive oxygen species upon photoactivation enabled effective bacterial cell killing. Spectroscopic studies confirmed strong binding of the conjugate to DNA through intercalation, likely disrupting DNA replication and transcription. Interaction studies with bovine serum albumin demonstrated substantial serum protein binding, which may positively impact the pharmacokinetics and biodistribution. Overall, this porphyrin-polymer conjugate offers a multifunctional theranostic platform, combining antimicrobial action with DNA and protein binding potential, positioning it as a promising candidate for aPDT and bioimaging applications.

Amyloid-Like Protein-Modified Carbon Nitride as a Bioinspired Material for Enhanced Photocatalytic CO2 Reduction.

Modification of g-C3N4 with metal-free biomaterials through an environmentally friendly, low-energy, facile, and rapid single-step method is desired for the preparation of photocatalysts with efficient activity and high selectivity of CO2 reduction but remains a great challenge. Herein, we develop a phase-transitioned protein modification strategy for photocatalysts through superfast amyloid-like protein assembly on surfaces using a one-step sequential coating method. Metal-free carbon nitride/protein heterojunction composite photocatalysts (the phase-transitioned lysozyme (PTL), phase-transitioned bovine serum albumin (PTB), and phase-transitioned ovalbumin (PTO)-coated carbon nitride@SiO2 (CN@SiO2) and bioinspired carbon nitride hollow nanospheres (CN-HS) obtained by etching of CN@SiO2) are prepared using lysozyme, bovine serum albumin, and ovalbumin. The insulator-semiconductor heterojunctions formed at the protein-carbon nitride interface promote the migration and separation of photogenerated charges. The exposed hydrophobic alkyl and aryl groups of the surface-modified protein enable the formation of a CO2-aqueous solution-photocatalyst three-phase interface on the catalyst surface and the exposed -NH2 groups provide sites for CO2 adsorption, which effectively increases CO2 mass transfer and its adsorption as well as hydrophobicity, promoting CO2 reduction and inhibiting hydrogen production. Therefore, protein modification effectively improves the CO2 reduction activity and CO selectivity. For instance, compared to CN-HS, the CO yield of the PTL-modified CN-HS (1346.5 μmol g-1) increased by 24.5 times and the CO selectivity reached 90.5%. These findings represent a critical advancement in the surface modification of carbon nitride for CO2 reduction and the design of bioinspired materials for various applications.

Oxygen consumption measurements at ultra-high dose rate over a wide LET range.

The role of radiolytic oxygen consumption for the in-vitro "Ultra-High Dose Rate" (UHDR) sparing and in-vivo FLASH effect is subject to active debate, but data on key dependencies such as the radiation quality are lacking. The influence of "dose-averaged Linear Energy Transfer" (LETd) and dose rate on radiolytic oxygen consumption was investigated by monitoring the oxygen concentration during irradiation with electrons, protons, helium, carbon, and oxygen ions at UHDR and "Standard Dose Rates" (SDR). Sealed "Bovine Serum Albumin" (BSA) 5% samples were exposed to 15Gy of electrons and protons, and for the first time helium, carbon, and oxygen ions with LETd values of 1, 5.4, 14.4, 65, and 100.3keV/µm, respectively, delivered at mean dose rates of either 0.3-0.4Gy/s for SDR or approximately 100Gy/s for UHDR. The Oxylite (Oxford Optronics) system allowed measurements of the oxygen concentration before and after irradiation to calculate the oxygen consumption rate. The oxygen consumption rate was found to decrease with increasing LETd from 0.351mmHg/Gy for low LET electrons to 0.1796mmHg/Gy for high LET oxygen ions at SDR and for UHDR from 0.317 to 0.1556mmHg/Gy, respectively. A higher consumption rate for SDR irradiation compared to the corresponding UHDR irradiation persisted for all particle types. The measured consumption rates demonstrate a distinct LETd dependence. The obtained dataset, encompassing a wide range of LETd values, could serve as a benchmark for Monte Carlo simulations, which may aid in enhancing our comprehension of oxygen-related mechanisms after irradiations. Ultimately, they could help assess the viability of different hypotheses regarding UHDR sparing mechanisms and the FLASH effect. The found LETd dependence underscores the potential of heavy ion therapy, wherein elevated consumption rates in adjacent normal tissue offer protective benefits, while leaving tumor regions with generally higher "Linear Energy Transfer" (LET) vulnerable.

Energy landscape of conformational changes for a single unmodified protein

Resolving the free energy landscapes that govern protein biophysics has been obscured by ensemble averaging. While the folding dynamics of single proteins have been observed using fluorescent labels and/or tethers, a simpler and more direct measurement of the conformational changes would not require modifications to the protein. We use nanoaperture optical tweezers to resolve the energy landscape of a single unmodified protein, Bovine Serum Albumin (BSA), and quantify changes in the three-state conformation dynamics with temperature. A Markov model with Kramers’ theory transition rates is used to model the dynamics, showing good agreement with the observed state transitions. This first look at the intrinsic energy landscape of proteins provides a transformative tool for protein biophysics and may be applied broadly, including mapping out the energy landscape of particularly challenging intrinsically disordered proteins.

Crystal Structures and Biological Profiles of Novel Tridentate Schiff Bases Nickel (II) Complexes

ABSTRACTNovel bioactive ternary nickel (II) complexes, [Ni(5BrSal‐Phe)(phen)] (1) and [Ni(5BrSal‐Tyr)(phen)] (2) (5BrSal‐Tyr: Schiff base derived from 5‐bromosalicylaldehyde and L‐tyrosine, 5BrSal‐Phe: Schiff base derived from 5‐bromosalicylaldehyde and L‐phenylalanine, phen: 1,10‐phenanthroline), have been synthesized and characterized by electronic absorption spectroscopy, CHN, FTIR, ESI‐MS and X‐ray crystallography techniques. Interaction of the complexes with biomolecules (calf thymus DNA (CT‐DNA) and bovine serum albumin (BSA)) has been investigated by electronic absorption and fluorescence spectroscopy. The results show that the complexes can bind to CT‐DNA via a minor groove binding mode. Moreover, the fluorescence quenching mechanism between the complexes and BSA is a static quenching process. The antiproliferative activities of the complexes against breast cancer cells (MCF‐7 and MDA‐MB‐231) and healthy breast epithelial cells (MCF‐10A) were investigated. The complex 2 was found to have promising antiproliferative activity in selected cell line, with lower IC50 values than cisplatin. Molecular docking studies suggest that the complexes may serve as potential chemotherapeutic agents. These complexes have been observed to interact with various targets within cells, including the epidermal growth factor receptor (EGFR), bovine serum albumin (BSA), and B‐DNA. Analyses indicate that these interactions are supported not only by conventional hydrogen bonds but also van der Waals forces and π‐π interactions. Additionally, determining binding constants and regions enhances our understanding of how the complexes interact with target molecules. The results emphasize the importance of the complexes in cancer therapy by highlighting the necessity of understanding their molecular‐level interactions with targets.

Optimization of Antiproliferative Properties of Triimine Copper(II) Complexes.

Cu(II) complexes with 2,2':6',2″-terpyridines (terpy) and 2,6-bis(thiazol-2-yl)pyridines (dtpy) with 1- or 2-naphtyl and methoxy-naphtyl were synthesized to elucidate the impact of the triimine core, naphtyl linking mode, and presence of methoxy groups on the antiproliferative activity of [CuCl2(Ln)]. Their antiproliferative effect was analyzed in ovarian (A2780) and colorectal (HCT116) carcinomas and colorectal carcinoma resistant to doxorubicin (HCT116-DoxR) cell lines and in normal human fibroblasts. Among all complexes, the 1- and 2-naphtyl substituted terpy Cu(II) complexes (Cu1a and Cu1b) showed the strongest cytotoxicity, namely, in HCT116-DoxR 2Dcells and were also capable of inducing the loss of cell viability in 3D HCT116-DoxR spheroids. Their intracellular localization, capability to increase reactive oxygen species (ROS), and interaction with DNA (nonintercalative mode) trigger oxidative DNA cleavage leading to cell death by apoptosis and autophagy. Cu1a and Cu1b do not show in vivo toxicity in a chicken embryo and can interact with bovine serum albumin (BSA).