Assay and drug development technologies最新文献

The utilization of herbal bioactive compounds for health maintenance is now increasing the interest of consumers because it has therapeutic benefits. Luteolin (LLN) is a natural bioactive compound and is found in various plant sources. It has many pharmacological activities, i.e., anticancer, antidiabetic, antioxidant, anti-inflammatory, and antimicrobial. It has poor water solubility, leading to low dissolution, low bioavailability, and low therapeutic efficacy. The present research work was to develop the LLN-loaded gel microbeads using a combination of sodium alginate (SA) and gum tragacanth polymers to strengthen microbeads (BD) and enhance the therapeutic efficacy. The microbeads were prepared by using the ionotropic gelation method and evaluated by various physicochemical parameters, i.e., particle size, encapsulation efficiency, swelling index, FITR, and X-ray diffraction study. The optimized microbeads (LLNBD3) showed a 97.63 ± 3.12% yield, 845 ± 6.21 μm in size, and 78.54 ± 3.65% drug entrapment efficiency. The microbeads exhibited excellent swelling in intestinal pH (6.8) compared with an acidic medium (pH 1.2). The LLNBD3 exhibited a sustained release profile (89.23 ± 2.51% in 12 h) with first-order release kinetics (R² = 0.9752) with the Fickian diffusion mechanism of drug release. The Fourier transform infrared spectra and X-ray diffractograms did not show any distinct peaks of LLN, revealing that the LLN was encapsulated into a microbeads matrix. The LLNBD3 showed significant antioxidant activity compared with pure LLN, confirmed by the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) method. In addition, it also showed remarkable in vitro anticancer activity against the colorectal cell line (HT-29) and antimicrobial activity against Staphylococcus aureus and Escherichia coli. The stability study demonstrated no significant change in swelling and release behavior. The finding concluded that tragacanth gum and SA microbeads could be promising drug carriers to improve the dissolution and oral delivery of herbal bioactive compounds and synthetic drugs.

Alzheimer's disease (AD) is a neurological disorder that results in the loss of memory and cognitive functions linked to redox disbalance, neuroinflammation, neurotransmitters changes, and the accumulation of amyloid-beta (1-42) plaques in AD. In this study, rats were administered with intracerebroventricular (ICV) streptozotocin (STZ) to produce AD-like symptoms in rats. ICV-STZ bilaterally, 3 mg/kg, was infused on days 1 and 3 with the help of Hamilton syringe by fixing cannula at the target position of rat brain using coordinates -2 mm (anteriposterior), 1.6 mm Mediolateral (ML), and 1.5 mm (dorsoventral). Learning and spatial memory were checked using Morris water maze and elevated plus maze apparatus. In ICV-STZ, rats lost their spatial and learning memory, increased level of prooxidant like Lipid peroxidation (LPO), nitrite and reduced glutathione (GSH), catalase, and superoxide dismutase (SOD) level. The increased level acetylcholinesterase (AChE) catalyzed acetylcholine (ACh) concentration indicates cholinergic neuron degeneration. Furthermore, we found raised inflammatory markers and altered neurotransmitters level after ICV-STZ. Administration of aescin (10, 20, and 30 mg/kg, p.o.) dose-dependently ameliorated the behavioral alteration and inhibited inflammatory markers like tumor necrosis factor-alpha, interleukin-6 (IL-6), and IL-1β. Furthermore, aescin restored antioxidants like GSH, SOD, and catalase and reduced the nitrite and lipid peroxidation level. AChE enzyme causes degradation of ACh, and its level was declined after treatment with aescin. Aescin also restored GABA, norepinephrine, and serotonin level in the brain with prevention of raised glutamate level. Moreover, the histopathological study confirmed neuronal pathogenesis, and aescin significantly achieved neuroprotective effect via preventing neuroinflammation, balancing redox potential, and inhibiting AChE enzyme.

RNA interference through small interfering RNA (siRNA) has shown great promise as a potential cancer treatment strategy in recent years. However, the delivery of siRNA to target cancer cells efficiently remains a significant challenge. This review aims to highlight the recent advances in nanotechnology-enabled siRNA delivery for cancer treatment, bridging the gap between bench research and clinical application. A comprehensive literature search was conducted to identify recent studies focused on the utilization of nanotechnology for siRNA delivery in cancer treatment. Key databases, including PubMed, Scopus, and Web of Science, were used, and relevant articles were screened. Several nanotechnology-based platforms for siRNA delivery have emerged in recent years, providing enhanced selectivity, improved stability, and controlled release profiles. The primary types of nanocarriers discussed include lipid-based nanoparticles, inorganic nanoparticles, polymeric nanoparticles, and exosomes. Nanotechnology-based siRNA delivery systems represent a promising avenue for cancer treatment. Although significant progress has been made in preclinical studies, translating these findings to clinical applications poses several challenges, including scale-up production, safety, and targeted delivery. Nevertheless, the recent developments in this field hold great promise in revolutionizing cancer therapy, providing hope for more effective and personalized treatment options in the future.

This study aimed to enhance the therapeutic efficacy of prednisolone by developing a nanostructured lipid carrier (NLC) system for topical ocular administration and addressing the limitations of current topical therapy. Drug-loaded NLCs (prednisolone acetate [PSA1]-PSA13) were formulated using high-pressure homogenization techniques, optimized with a central composite design, and evaluated for various pharmaceutical properties. An optimization study indicates that the lipid ratio and surfactant concentration influence particle size, entrapment efficiency (EE), and drug release. The optimized NLC formulation (PSA3) was integrated into a pH-triggered in situ gel system and assessed for drug permeation, ocular irritation, and stability. The optimized drug-loaded NLC formulations (PSA3) exhibited a nano size of 96.80 ± 0.51 nm, achieving an EE of 84.51 ± 1.31% and a drug release rate of 95.76 ± 1.23%. The drug permeation through the goat cornea was significantly higher in the in situ gel (PSAG4) compared with the control (marketed PSA eye drop). Additionally, the eye irritation data indicated good ocular tolerance, while stability studies confirmed that the developed formulation remained stable at room temperature. In conclusion, the developed NLC-based in situ gel appears to be a promising approach to enhancing the efficacy of prednisolone in topical therapy for the successful treatment of ocular inflammation.

Stroke is an intricate oxidative and inflammatory response resulting from cerebral ischemia followed by reperfusion injury. Complex pathophysiology of stroke poses challenges for treatment. Peroxisome proliferator-activated receptor (PPAR) expression in the rat hippocampus is markedly elevated post cerebral ischemia/reperfusion (I/R) injury. Hence, saroglitazar, a dual PPAR-α/γ agonist, was investigated against cerebral I/R injury in rats. Male Sprague-Dawley rats were subjected to bilateral common carotid artery occlusion for 30 min and reperfusion for 3 days. During the reperfusion, animals were treated with vehicle or saroglitazar once a day for 3 days. The behavioral parameters were assessed, and animals were sacrificed to measure oxidative markers (malondialdehyde, superoxide dismutase, catalase, and reduced glutathione), inflammatory markers (interleukin-6, tumor necrosis factor-α, nuclear factor kappa-light chain enhancer of activated B cells (NF-κB), and high mobility group box 1 (HMGB-1) protein, infarction, and histopathology changes. Following I/R injury, antioxidant enzymes were reduced, while nitric oxide and inflammatory markers were increased in the disease group. In the rat hippocampus, these changes led to neurobehavioral impairment and cerebral infarction. Saroglitazar improved the levels of antioxidants and reduced inflammation; 2,3,5-triphenyltetrazolium chloride stain and histopathological analysis revealed the neuroprotective effect of saroglitazar in the hippocampus region. The neuroprotective effects of saroglitazar were attributed to its activation of both PPAR-α and PPAR-γ. It improved antioxidant levels and inhibited proinflammatory cytokines by suppressing the HMGB-1/NF-κB signaling pathway. These findings underscore the potential of saroglitazar in mitigating cerebral I/R injury.

Methotrexate (MTX) is an effective anticancer agent with limited water solubility, resulting in lower absorption in the gastrointestinal tract when administered orally. The present aim of the study is to construct sustained-release formulation of MTX-loaded microsponges with enhanced intestinal absorption and bioavailability using a quasi-emulsion solvent diffusion method. The Box-Behnken design (BBD) was adopted for this purpose. Particle size, encapsulation efficiency (EE), Q 2 h % (% drug release in 2 h), and Q 24 h % (% drug release in 24 h) were used as dependent factors, and polyvinyl alcohol, solvent, and stirring speed were used as independent factors. The prepared microsponges were characterized to assess their particle size and encapsulation efficacy (%). Attenuated total reflectance-Fourier transform infrared spectroscopy and differential scanning calorimetry were used to verify the compatibility study. Moreover, the cytotoxicity study was conducted on the HT-29 cell line. The optimized formulation exhibited a % encapsulation efficacy of 87.191% and a particle size of 2.176 µm. Furthermore, the optimized formulation demonstrated sustained drug release (85.71%) in Simulated Gastric Fluid (SGF) fluid at different pHs 1.2, 6.8, and 7.4. The stability study of the optimized formulation revealed good stability in terms of drug release, % encapsulation efficacy, and particle size. The results of the optimized formulation demonstrated that the viability of HT-29 colon cancer (CC) cells was dose-dependently decreased by MTX-loaded microsponges. BBD was successfully employed for the development and optimization of MTX microsponges filled in Eudragit S-100-coated hard gelatin capsule, depicting their potential release of MTX from microsponges capsule only at the colonic region and found to be potential carrier system for CC.

Cytotoxicity assays are essential in the field of research as they enable the examination of cellular responses to stimuli and shed light on complex mechanisms involved in multiple diseases and drug development. This review covers a range of cytotoxicity assays, including trypan blue and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assays, to more advanced techniques like caspase activity assays, Lactate dehydrogenase release assays, comet assays, and micronucleus assays for DNA damage assessment. Apart from these, other relevant assays like Alamar Blue, Bromodeoxyuridine incorporation, and clonogenic cell survival are also discussed. In this study, significance of these assays in drug development, toxicology studies, and biomedical research is discussed in detail, highlighting their role in ensuring safety and unraveling disease mechanisms. Furthermore, we explore emerging technologies such as chip-based assays, organ-on-a-chip systems, and high-throughput screening, which enhance precision and efficiency in research. Despite these advancements, challenges remain that necessitate standardization efforts and the development of more refined models. In conclusion, this review reflects on the evolving landscape of cytotoxicity assays, finding a balance between traditional methodologies and cutting-edge technologies.

Buzhong Yiqi decoction (BZYQD) is a traditional Chinese medicine prescription for treating neurogenic bladder (NB). However, the underlying pharmacological mechanism remains unclear. This study aims to clarify the related molecular mechanism. Molecular structure information and targets of core components of BZYQD were obtained from Traditional Chinese Medicines Systems Pharmacology Platform (TCMSP) and SwissTargetPrediction databases. Genes involved in NB were obtained from Comparative Toxicogenomics Database, DisGeNet, GeneCards, and Online Mendelian Inheritance in Man databases. The hub targets of BZYQD in NB treatment were identified by protein-protein interaction (PPI) network analysis with STRING platform and analyzed by gene ontology analysis and the Kyoto Encyclopedia of Genes and Genomics pathway enrichment analysis. Molecular docking was used to verify the binding affinity between the hub targets and the bioactive components of BZYQD. Subsequently, the neuroprotective and anti-inflammatory effects of main bioactive components of BZYQD were investigated with in vitro assays. A total of 131 candidate compounds and 925 predicted target genes were screened. PPI network analysis suggested that ESR1, EGFR, HSP90AA1, MAPK3, AKT1, and CASP3 were the hub targets. BZYQD treatment was associated with hypoxia inducible factor-1 (HIF-1) signaling pathway. Dehydroglyasperin C (DGC), N-cis-feruloyltyramine, shinpterocarpin (SHI), gancaonin M, and glyasperin B, as the main bioactive components of BZYQD, had good binding affinity with hub target proteins. DGC and SHI treatment could significantly inhibit the injury of neurons and inflammatory response of microglia stimulated by oxidized low-density lipoprotein (ox-LDL), respectively. In summary, BZYQD and its main bioactive components DGC and SHI show good potential to ameliorate the symptoms of NB.

The present study highlighted enhancing the therapeutic effectiveness of curcumin (CUR) co-loaded lawsone (LS) through a solid lipid nanoparticles (SLNs)-based delivery system. The cetyl palmitate (CP), polyethylene glycol 400 (PEG), and probe sonication time (PS) were considered as independent variables whereas particle size and % entrapment efficiency (EE) were selected as dependent variables. The CUR-LS-SLN was developed by hot emulsification followed by probe sonication. A 2³ factorial design was utilized in formulation development using JMP software version 17. Notably, the particle size and %EE of all the formulations were about 500 nm and greater than 75%, respectively. The zeta potential value was found to be -46.8 mV. From leverage plots significant and sensitive factors on particle size and %EE were identified. Contour plots led to the identification of an optimized formula whereby maintaining CP at 100 mg, PEG 400 at 6 mL, and PS at 10 min the desired particle size and %EE was achieved. TEM studies indicated the spherical shape of the particles. MTT assays of Michigan Cancer Foundation-7 (MCF-7) cells showed enhanced efficacy and greater cell inhibition of CUR-LS-SLN and combining both drugs using nanocarriers gave superior inhibition as compared with using either of the drugs evident from IC₅₀ values of 3.7, 9.4, and 2.5 μM, respectively, for CUR, LS, and CUR-LS-SLN. The cells in the combination mostly had irregular cell walls and cell shrinkage was noted and greater cell reduction was also seen. It was found that the enhanced cytotoxicity effect of MCF-7 cells on the developed formulation was attributed to the drug's synergistic actions, more efficient nanocarrier internalizations, and sustained drug release from the formulation. Stability studies indicated that the optimized SLN was stable for 6 months.