Caco-2 Cells Research Articles

Antiproliferative Activity of Antioxidants-Rich Extracts from Olea europaea L. Leaves and Oil Against Six Cancerous Human Cell Lines

ABSTRACT The present work aims to study the cytotoxic effect of the methanolic extract of leaves and oil of three olive cultivars: Manzanillo, Picual and Koronieki. Against six cancer cell lines, namely MCF-7 and MDA (breast cancer), HepG2 (liver cancer), PC3 (prostate cancer), Caco-2 (colon cancer), and A549 (lung cancer) using the MTT method. The results obtained showed that Manzanillo leaf methanolic extract (LMME) and Manzanillo oil (OMME) gave the highest cytotoxicity with the lowest IC50 against Caco2 and A549 cell lines. To explain this result, the mechanism of action was studied at the chemical, cellular and molecular levels. The chemical composition analysis of LMME and OMME was performed by GC-MAS. Furthermore, phytochemical analysis was performed. In addition to cell cycle analysis and apoptosis induction, the gene expression of three apoptotic genes was measured. The total phenolic contents (TPC) and total flavonoid contents (TFC) of (LMME) were greater than that of (OMME). Both LMME and OMME had arrested cell cycle at G1 phase for each of Caco2 and A549 cells. LMME and OMME induce up regulation of Casp3 and BAX genes and down regulation of Bcl-2 gene. These results indicated that, LMME and OMME had anti-proliferation effect against Caco2 and A549 cells through induction of apoptosis pathway. Further detailed studies are needed to examine the effect of each fraction of whole extract.

Chitooligosaccharide-modified PLGA-loaded PPD nanoparticles ameliorated sepsis-associated acute kidney injury via the NF-κB signaling pathway

Objectives Sepsis-associated acute kidney injury (SA-AKI) is a significant clinical challenge with high morbidity and mortality. Low bioavailability of protopanaxadiol (PPD) limits its clinical application. In this study, PPD was encapsulated with chitooligosaccharide (COS) modified polylactic-co-glycolic acid (PLGA) to develop novel nanomedicines for the treatment of SA-AKI. Methods COS-PLGA-PPD nanoparticles were prepared by emulsified solvent evaporation method, and their properties were evaluated. In vitro, the anti-inflammatory and protective effects of COS-PLGA-PPD NPs were investigated in a cellular model of LPS-induced NRK-52E cells and their uptake in Caco-2 cells. Indicators of renal injury, inflammation, and NF-κB signaling pathway were evaluated by injecting LPS into SD rats and inducing SA-AKI model in vivo. The oral bioavailability of nanoparticles was evaluated by pharmacokinetics. Results Compared with PPD and unmodified nanoparticles, COS-PLGA-PPD NPs were more stable, with a particle size of 139.69 nm, which enhanced the viability of NRK-52E cells, increased the uptake of Caco-2 cells, alleviated the symptoms of SA-AKI in rats, inhibited the NF-κB signaling pathway, reduced the expression of inflammatory factors, and had a bioavailability 1.7-fold that of PPD. Conclusion COS-PLGA-PPD NPs ameliorate LPS-induced SA-AKI in rats by inhibiting the NF-κB signaling pathway, providing a basis for the treatment of SA-AKI.

Computational studies for pre-evaluation of pharmacological profile of gut microbiota-produced gliclazide metabolites

BackgroundGliclazide, a second-generation sulfonylurea derivative still widely used as a second-line treatment for type 2 diabetes mellitus, is well known to be subject to interindividual differences in bioavailability, leading to variations in therapeutic responses among patients. Distinct gut microbiota profiles among individuals are one of the most crucial yet commonly overlooked factors contributing to the variable bioavailability of numerous drugs. In light of the shift towards a more patient-centered approach in diabetes treatment, this study aimed to conduct a pharmacoinformatic analysis of gliclazide metabolites produced by gut microbiota and assess their docking potential with the SUR1 receptor to identify compounds with improved pharmacological profiles compared to the parent drug.MethodsTen potential gliclazide metabolites produced by the gut microbiota were screened for their pharmacological properties. Molecular docking analysis regarding SUR1 receptor was performed using Molegro Virtual Docker software. Drug-likeness properties were evaluated using DruLiTo software. Subsequently, the physicochemical and pharmacokinetic properties of gliclazide and its metabolites were determined by using VolSurf+ software package.ResultsAll studied metabolites exhibited better intrinsic solubility than gliclazide, which is of interest, considering that solubility is a limiting factor for its bioavailability. Based on the values of investigated molecular descriptors, hydroxylated metabolites M1-M6 showed the most pronounced polar and hydrophilic properties, which could significantly contribute to their in vivo solubility. Additionally, docking analysis revealed that four hydroxyl-metabolites (M1, M3, M4, and M5), although having a slightly poorer permeability through the Caco-2 cells compared to gliclazide, showed the highest binding affinity to the SUR1 receptor and exhibited the most suitable pharmacological properties.ConclusionIn silico study revealed that hydroxylated gut microbiota-produced gliclazide metabolites should be further investigated as potential drug candidates with improved characteristics compared to parent drug. Moreover, their part in the therapeutic effects of gliclazide should be additionally studied in vivo, in order to elucidate the role of gut microbiota in gliclazide pharmacology, namely from the perspective of personalized medicine.

Isolation and in vitro investigation on lactic acid bacteria for potential probiotic properties from cat feces

BackgroundProbiotics, which are beneficial to the host, have been shown to benefit the health of cats. Lactic acid bacteria (LAB) are commonly used probiotics, but most strains used for cats are not derived from cats, leading to reduced efficacy and poor adaptation to cats. The objective was to identify LAB with promising probiotic potential specific to cats.MethodLABs were isolated from fecal samples of 20 healthy cats. Gram staining and the survival rate in the simulated gastrointestinal tract were used for preliminary screening. Candidate strains were identified by 16S rDNA sequencing, and further evaluated for adhesion ability, growth characteristics, antibacterial activity, antioxidant capacity, and safety.Results24 Gram-positive isolates were identified, with 10 (F1-F10) showing robust viability in the simulated gastroenteric fluid. These 10 strains exhibited excellent adhesion to Caco-2 cells and strong auto-agglutination properties. They also possessed the capacity to antagonize and aggregate pathogens (Staphylococcus aureus ATCC 25923, Salmonella Braenderup H9812, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa PAO1), Moreover, all strains demonstrated tolerance to H2O2 concentrations ranging from 0.5–2 mmol/L and the ability to scavenge 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radicals, indicating a certain level of antioxidant activity. Safety tests showed no hemolytic activity, and all but F6 were highly sensitive to antibiotics, with over 62.5% sensitivity to 16 antibiotics. Remarkably, F4 (Lactobacillus reuteri) and F10 (Lactobacillus brevis) exhibited exceptional viability in the simulated gastrointestinal tract, coupled with robust growth potential, enhanced adhesion efficiency, significant antibacterial and antioxidant properties.ConclusionOur findings revealed that F4 (Lactobacillus reuteri) and F10 (Lactobacillus brevis) hold promising potential as probiotics. This research lays a solid scientific foundation for the selection and application of probiotics tailored specifically for cats.

Colon-Targeted Poly(ADP-ribose) Polymerase Inhibitors Synergize Therapeutic Effects of Mesalazine Against Rat Colitis Induced by 2,4-Dinitrobenzenesulfonic Acid

Background/Objectives: In addition to oncological applications, poly(ADP-ribose) polymerase (PARP) inhibitors have potential as anti-inflammatory agents. Colon-targeted delivery of PARP inhibitors has been evaluated as a pharmaceutical strategy to enhance their safety and therapeutic efficacy against gut inflammation. Methods: Colon-targeted PARP inhibitors 5-aminoisoquinoline (5-AIQ) and 3-aminobenzamide (3-AB) were designed and synthesized by azo coupling with salicylic acid (SA), yielding 5-AIQ azo-linked with SA (AQSA) and 3-AB azo-linked with SA (ABSA). Additional conjugation of AQSA with acidic amino acids yielded glutamic acid-conjugated AQSA (AQSA-Glu) and aspartic acid-conjugated AQSA, which further increased the hydrophilicity of AQSA. Results: The distribution coefficients of PARP inhibitors were lowered by chemical modifications, which correlated well with drug permeability via the Caco-2 cell monolayer. All derivatives were effectively converted to their corresponding PARP inhibitors in the cecal contents. Compared with observations in the oral administration of PARP inhibitors, AQSA-Glu and ABSA resulted in the accumulation of much greater amounts of each PARP inhibitor in the cecum. ABSA accumulated mesalazine (5-ASA) in the cecum to a similar extent as sulfasalazine (SSZ), a colon-targeted 5-ASA prodrug. In the DNBS-induced rat colitis model, AQSA-Glu enhanced the anticolitic potency of 5-AIQ. Furthermore, ABSA was more effective against rat colitis than SSZ or AQSA-Glu, and the anticolitic effects of AQSA-Glu were augmented by combined treatment with a colon-targeted 5-ASA prodrug. In addition, the colon-targeted delivery of PARP inhibitors substantially reduced their systemic absorption. Conclusions: Colon-targeted PARP inhibitors may improve the therapeutic and toxicological properties of inhibitors and synergize the anticolitic effects of 5-ASA.

The lower content of mineral-complexing compounds favored the in vitro and in vivo iron bioavailability of biofortified cowpeas

This study aimed to evaluate in vitro and in vivo iron bioavailability of biofortified cowpeas (Vigna unguiculata (L) Walp) (Aracê, Xiquexique and Tumucumaque), compared to a conventional cultivar (Pajeú). In vitro bioavailability was evaluated in Caco-2 cells. For the in vivo study, 40 male Wistar rats were induced to anemia during 21 days (depletion phase), then, in repletion phase (14 days), the animals received as iron source (12 ppm): ferrous sulfate; Pajeú; Xiquexique, Aracê; or Tumucumaque. Pajeú had higher levels of complexing compounds than the biofortified cultivars. In Caco-2 cells, the cellular iron uptake index was higher for biofortified cowpeas, particularly Tumucumaque. Concerning the in vivo study, Tumucumaque group had the best hemoglobin regeneration efficiency and relative biological value, and improved colon crypt size. Then, the iron bioavailability was higher in the biofortified cowpeas, especially Tumucumaque, possibly due to the lower content of mineral-complexing compounds and improvement of absorptive area. This work provides important information to support strategies to combat iron deficiency.

Comprehensive evaluation of Capsosiphon fulvescens water extract: Assessing its effects on intestinal barrier integrity and inflammation in vitro and in vivo

Capsosiphon fulvescens has potential benefits for functionality. However, research on its impact on gut health is limited. In this study, the therapeutic efficacy of Capsosiphon fulvescens water extract (CFW) was investigated, with its intestinal health benefits confirmed in Caco-2 cell and mouse models. In the animal study, 32 male C57BL/6 mice were divided into control, DSS, and CFW treatment groups and treated for 28 days, with DSS given ad libitum in 7 days. Significant improvements in various biomarkers related to intestinal barrier function, including increased transepithelial electrical resistance values and reduced permeability, were observed in vitro model. Moreover, the administration of CFW was associated with improved histological scores and improved levels of TJ proteins and cytokines. The therapeutic efficacy of CFW is believed to be associated with the bioactive compounds found in the extract. The findings here highlight the promising therapeutic potential of CFW for managing inflammatory bowel disease.

Are Nanostructured Lipid Carriers (NLC) better than Solid Lipid Nanoparticles (SLN) for delivering abiraterone acetate through the gastrointestinal tract?

Abiraterone acetate (AbA) is a progesterone derivative indicated for the treatment of metastatic prostate cancer. This BCS (Biopharmaceutics Classification System) Class IV molecule has an extremely poor oral bioavailability (<10 %), notably due to its very low water solubility and intestinal permeability. Among the few existing galenic strategies to improve AbA’s oral bioavailability, lipid nanoparticles such as Solid Lipid Nanoparticles (SLN) and Nanostructured Lipid Carriers (NLC) are relevant nanovectors. The objective of this study is to develop and compare SLN and NLC for oral delivery of abiraterone acetate. Both SLN and NLC are biocompatible, biodegradable and produced by high pressure homogenization (HPH), an ecological-friendly manufacturing process, organic solvent-free and easily scalable. The HPH process allowed the formation of AbA-loaded SLN and NLC with particle size lower than 160 nm and high encapsulation efficiencies. The addition of a liquid lipid significantly reduced the mean diameter of the nanoparticles, reflecting the greater benefit of the NLC formulation compared to SLN. Both SLN and NLC formulations offered an important protection of AbA in intestinal media, with a better stability for NLC. When encapsulated in SLN or NLC, the AbA is strongly retained by the nanoparticles, whatever the dissolution medium, which means that both formulas are able to protect and retain the drug in the intestinal tract, right up to its delivery to the enterocytes surface. High concentrations of nanoparticles were administered without cytotoxicity, especially for the NLC, which provides a real added value in terms of biocompatibility with Caco-2 cells. Finally, the nanoparticles were able to penetrate into enterocytes by the transcellular route, demonstrating an intense cellular internalization.

Encapsulation of yarrow phenolic compounds in lupin protein nanoemulsions increases stability during gastrointestinal transit and delivery in the colon

This study aimed to assess the behaviour of phenolic compounds from yarrow extract encapsulated in nanoemulsions during in vitro gastrointestinal digestion. Oil-in-water nanoemulsions were developed using grape seed oil and lupin protein (LPI) as oil phase and emulsifier, respectively. The use of 6 % LPI including 1 mg/mL of yarrow extract resulted in nanoemulsions with a homogeneous particle size distribution (200 nm) and an encapsulation efficiency of 85.6 %. During in vitro gastrointestinal digestion, most of the phenolics remained encapsulated, being protected from degradation. The in vitro bioavailability of the encapsulated phenolics was measured using a cell co-culture model (Caco-2/HT-29MTX). In this regard, nanoemulsions did not increase the bioavailability of yarrow phenolics, instead, they promoted their access to the colon. Finally, the antiproliferative activity was determined in Caco-2 cells, observing that the apical fraction inhibited cancer cells, indicating the bioefficacy of the non-absorbed phenolics.Thus, this study underscores the potential of LPI-stabilized nanoemulsions as a vehicle for protecting and delivering yarrow phenolics to the colon.

Theabrownin/whey protein isolate complex coacervate strengthens C2C12 cell proliferation via modulation of energy metabolism and mitochondrial apoptosis.

Theabrownin (TB)-whey protein isolate (WPI) complex coacervates (TW) were firstly prepared to investigate the regulatory effects on skeletal muscle. The binding of TB to WPI reached saturation with the strongest electrostatic interaction at the ratio of 10:1. The formation of TW was driven by electrostatic interactions with the aid of hydrogen bonding and hydrophobic interactions, and the digestion behavior of TW was investigated based on in vitro gastrointestinal and CaCO2 cell models. The regulatory effect of TW on muscle cells was investigated by C2C12 cell assay. Cell cycle analysis showed that TW promoted the transition of skeletal muscle cells from proliferative state to differentiated state. Immunofluorescence and gene expression revealed that TW positively regulated myogenic regulatory factors, contributing to myofiber formation. Moreover, TW activated the intracellular TCA cycling and oxidative phosphorylation, providing energy for skeletal muscle regeneration and repair. Mechanistically, TW inhibited the release of cytochrome C from mitochondria to cytoplasm through the Bcl-2/Cytochrome C/Cleaved-Caspase-3 pathway, exhibiting a protective effect on skeletal muscle cells. In the future, the molecular mechanism of TW enhancing skeletal muscle function should be validated through aging animal models and clinical trials and expand its therapeutic application for muscle health in functional food and dietary supplements.

Functionalisation of chitosan with methacryloyl and crotonoyl groups as a strategy to enhance its mucoadhesive properties

Mucoadhesive polymers are crucial for prolonging drug retention on mucosal surfaces. This study focuses on synthesising and characterising novel derivatives by reacting chitosan with crotonic and methacrylic anhydrides. The structure of the resulting derivatives was confirmed using proton-nuclear magnetic resonance spectroscopy and Fourier-transform infrared spectroscopy. It was established that the degree of substitution plays a crucial role in the pH-dependent solubility profiles and electrophoretic mobility of the chitosan derivatives. Spray-drying chitosan solutions enabled preparation of microparticles, whose mucoadhesive properties were evaluated using fluorescence flow-through studies and tensile test, demonstrating improved retention on sheep nasal mucosa for modified derivatives. Acute toxicity studies conducted in vivo using planaria and in vitro using MTT assay with the Caco-2 cell line, a model of the mucosal epithelium in vitro, showed that the novel derivatives are not cytotoxic. These findings emphasise the potential of tailored chitosan chemical modifications for enhancing transmucosal drug delivery.

When subcellular chemical imaging enlightens our understanding on intestinal absorption, intracellular fate and toxicity of PFOA in vitro

Perfluorooctanoic acid (PFOA) is a persistent organic pollutant that accumulates in the human body, leading to major health issues. Upon oral uptake, the gastrointestinal tract is the first biological barrier against PFOA. However, the localization of PFOA and its impact on the intestinal wall are largely unknown. Here we achieve a breakthrough in the knowledge of intestinal absorption, intracellular fate and toxicity of PFOA using in vitro assays combined with novel analytical imaging techniques. For the first time, we localized PFOA in the cytosol of Caco-2 cells after acute exposure using high spatial resolution mass spectrometry imaging, and we estimated the PFOA cytosolic concentration. Knowing that PFOA enters and accumulates in the intestinal cells, we also performed common toxicity assays assessing cell metabolic activity, membrane integrity, oxidative stress response, and cell respiration. This study integrating powerful analytical techniques with widely used toxicology assays provides insightful information to better understand potential negative impacts of PFOA and opens new opportunities in toxicology and life science in general.

Bioavailability of encapsulated rosmarinic acid and β-Carotene in beef Sausages: A Caco-2 cell study

In this study, we examined the bioavailability and functionality of rosmarinic acid and β-carotene in beef sausages using Caco-2 cell models. Digesta from enriched sausages showed high antioxidant activity, with 89.2% DPPH and 87.65% ABTS inhibition. Micrographs revealed the impact of co-delivery during digestion, with significantly higher levels of rosmarinic acid in sample B (80% rice starch & 20% rice protein) and β-carotene in sample C (60% rice starch & 40% rice protein) compared to the control digesta, attributed to the rice-starch network ensuring targeted release. During the total absorption of sample C, rosmarinic acid was more bioavailable than β-carotene. The carrier material structure plays a crucial role in the release of bioactive compounds into the mixed micellar phase. Encapsulation and incorporation into suitable food matrices such as meat can enhance the bioavailability of rosmarinic acid and β-carotene, potentially leading to the production of healthier meat products. Caco-2 cells, which resemble human intestinal cells, were used to study the absorption and transport of bioactive compounds post-gastrointestinal digestion. In-depth knowledge of the digestibility of specific dietary matrices is required to understand the metabolic dynamics of these substrates, which ultimately affect their bioavailability. The creation of functional meat products using rosmarinic acid and β-carotene ensures their complete utilisation.

Bioavailability study of OTA, ZEN, and AFB1 along with bioactive compounds from tiger nut beverage and its by-products

Mycotoxins pose significant health risks due to their prevalence in food products and severe health implications, including carcinogenicity. This study investigates the bioavailability of mycotoxins aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN) individually and combined, in the presence of identified polyphenols from tiger nut beverage (TNB) and tiger nut by-product (TNBP) using the in vitro model Caco-2 cells, which simulates the human intestinal barrier. The objective is to understand how bioactive compounds from TNBP can mitigate the effects of AFB1, OTA and ZEN (and their combination) by bioavailability interference, contributing to safer food products and innovative food safety strategies. In vitro gastrointestinal digestion was simulated using the INFOGEST protocol, followed by a bioavailability assessment through transepithelial transport assays in differentiated Caco-2 cells. OTA bioavailability significantly increased in the presence of TNB and TNBP, suggesting interactions that enhance its intestinal absorption. AFB1 maintained high bioavailability across all conditions (up to 83%), while ZEN showed a general decrease (up to 24%), thus indicating a potential protective effect of TNB and TNBP against ZEN toxicity. Regarding the effect of mycotoxins on the bioavailability of polyphenols from TNB and TNBP, a general enhancement was observed for TNB consistently showing higher bioavailability than for TNBP. Notably, OTA and ZEN significantly increased polyphenols bioavailability, reaching up to 79.2% in TNB. Individual polyphenol generally showed a notable reduction in trans-ferulic acid and an increase in trans-cinnamic acid in the presence of mycotoxins. For TNBP, individual mycotoxins generally enhanced polyphenol bioavailability, with AFB1 showing the most significant increase. In conclusion, tiger nut products show promise as sources of bioactive compounds for mitigating mycotoxin toxicity in food products. However, further studies are necessary to clarify these interactions and optimize the conditions of use for their safe and effective application in the food industry.

Downregulated KLF4, induced by m6A modification, aggravates intestinal barrier dysfunction in inflammatory bowel disease

BackgroundKrüppel-like factor 4 (KLF4), a transcription factor, is involved in various biological processes. However, the role of KLF4 in regulating the intestinal epithelial barrier (IEB) in inflammatory bowel disease (IBD) and its mechanism have not been extensively studied.MethodsKLF4 expression in IBD patients and colitis mice was analyzed using Gene Expression Omnibus(GEO) database, immunohistochemistry (IHC) and Western blot. The roles of KLF4 in IEB and colitis symptoms were verified in dextran sulfate sodium (DSS)-induced and 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis model mice using an adenovirus carrying KLF4 shRNA (shKLF4-Adv). Furthermore, the influence of KLF4 on trans-epithelium electrical resistance (TEER), paracellular permeability, apical junction complex (AJC) protein expression and apoptosis was assessed in vitro and in vivo. MeRIP and RIP assays were used to verify the effects of m6A modification on KLF4 expression.ResultsKLF4 expression was significantly decreased in IBD patients and was negatively associated with inflammatory features. KLF4 deletion aggravated colitis symptoms and IEB injuries by reducing AJC protein expression and increasing apoptosis in mice with colitis. Furthermore, KLF4 transcriptionally regulated the expression of AJC proteins and inhibited apoptosis by reducing cellular ROS levels and proinflammatory cytokine expression. Moreover, we observed that METTL3/ALKBH5/YTHDF2-mediated m6A modification led to a decrease in KLF4 expression in Caco-2 cells. In addition, APTO-253, an inducer of KLF4, exhibited a synergistic effect with mesalazine on IEB function.ConclusionsOur study demonstrated that KLF4 is a crucial regulator of IEB, suggesting that targeting KLF4 may be a promising therapeutic alternative for IBD.

TPM4 overexpression drives colon epithelial cell tumorigenesis by suppressing differentiation and promoting proliferation

ObjectiveThe high morbidity and mortality associated with colorectal cancer (CRC) and the recent increases in early-onset CRC obviate the need for novel methods to detect and treat this disease, particularly at early stages. We hypothesize that aberrant expression of genes involved in the crypt-luminal migration of colon epithelial cells, a process necessary for their growth arrest and maturation, may disrupt differentiation and transition cells from a normal to tumorigenic state. MethodsWe searched for contractility- and motility-related genes that are dysregulated in human CRC relative to normal colon. RNA expression of one such gene, tropomyosin 4 (TPM4), was measured by qRT-PCR and RNA-seq in human colorectal tissues at various stages of tumorigenesis: CRC, adenoma, and at-risk (grossly normal mucosa from a patient with Familial Adenomatous Polyposis, or FAP), relative to controls. Effects of aberrant TPM4 expression on colon epithelial cell proliferation and maturation were determined by overexpression using stable transfection in spontaneously differentiating Caco2 cells or silencing using siRNA in proliferating cells. ResultsTPM4 is overexpressed at various stages of tumorigenesis, including CRC, adenoma, and grossly normal FAP colon tissue, as well as in proliferating versus differentiating Caco2 cells. TPM4.2 overexpression in differentiating Caco2 cells markedly inhibits certain aspects of maturation, notably sucrase isomaltase and glutathione-S-transferase alpha1 expression, and causes morphological and cell junction abnormalities. Conversely, siRNA-mediated suppression of TPM4.2 inhibits Caco2 proliferation. ConclusionsTPM4 overexpression attenuates colon epithelial cell differentiation and promotes proliferation. Therefore, TPM4 expression may be a biomarker to enhance strategies for CRC diagnosis and treatment.

Protective Effects of Spirulina Against Lipid Micelles and Lipopolysaccharide-Induced Intestinal Epithelium Disruption in Caco-2 Cells: In Silico Molecular Docking Analysis of Phycocyanobilin

Damage to intestinal epithelial cells is present in obesity and other diseases because of inflammatory and oxidative processes. This damage compromises the gastrointestinal barrier, killing enterocytes, altering intestinal permeability, and eliciting abnormal immune responses that promote chronic inflammation. Recent evidence shows that spirulina is a potent natural agent with antioxidant and anti-inflammatory properties. Objectives: This study was conducted to evaluate the effect of spirulina aqueous extract (SPAE) on the alterations of the intestinal epithelium induced by lipid micelles (LMs) and/or inflammation induced by lipopolysaccharides (LPSs) in the Caco-2 cell line. Methods: In the current research, we assessed the protective actions of SPAE against cytotoxicity, oxidative stress, inflammation, and epithelial barrier perturbation by using an in vitro model, the intestinal Caco-2 cells, treated with LPSs and/or LMs. We also performed an in silico molecular docking analysis with spirulina’s bioactive compound, phycocyanobilin. Results: Our results showed that SPAE has no cytotoxic effect on Caco-2 cells. On the contrary, it improved cell viability and exhibited anti-inflammatory and antioxidant actions. SPAE also protected against endoplasmic reticulum stress and tight junction proteins, thus improving the epithelial barrier. The in silico study revealed a strong binding affinity of the phycocyanobilin compound with human SOD and NADPH oxidase and a good binding affinity towards COX-2 and iNOS. Conclusions: Taken together, these findings demonstrate the beneficial actions of SPAE on Caco-2 cells, suggesting it may be useful in preserving the epithelial intestinal barrier in human conditions involving oxidative stress and inflammation such as obesity.

Effects of Forming Lactoferrin–Milk Protein Complexes on Lactoferrin Functionality and Intestinal Development in Infancy

Background/Objectives: Lactoferrin (Lf) is an iron-binding glycoprotein with multiple bioactivities, including promotion of cell proliferation and differentiation, immunomodulation, and antimicrobial activity. Lf, a basic glycoprotein, can bind to α-lactalbumin (α-Lac), an acidic whey protein. The current study aimed to evaluate whether Lf forms protein complexes with α-Lac and proteins/peptides from whey protein hydrolysate (WPH) and nonfat bovine milk powder (MP) and whether forming protein complexes influences resistance to gastrointestinal digestion and affects the bioactivities of Lf in human intestinal epithelial cells (HIECs and differentiated Caco-2 cells). Methods: Lf was blended with α-Lac, WPH, or MP. Assays were conducted to evaluate the bioactivities of proteins (Lf, α-Lac, WPH, or MP) and Lf–protein blends on HIECs and Caco-2 cells. Results: (1) Lf forms complexes with α-Lac and proteins/peptides from WPH and MP; (2) compared with Lf alone, complexed Lf shows greater resistance to in vitro digestion; (3) forming protein complexes does not affect Lf’s binding to the Lf receptor or its uptake by HIECs; and (4) forming protein complexes does not impact Lf’s bioactivities, including the promotion of cell proliferation and differentiation, reduction of cell permeability by upregulating tight-junction proteins, immune modulation through the regulation of IL-18, inhibition of enteropathogenic Escherichia coli growth, and modulation of immune responses to EPEC infection. Conclusions: Lf forms complexes with α-Lac and other milk proteins/peptides from WPH and MP in protein blends, and forming complexes does not affect the functionalities of Lf.

Thermoplastic Molding of Silk-Curcumin Sustainable Composite Materials with Antibacterial Properties.

Silk fibroin (SF) is a natural protein generated from the Bombyx mori silkworm cocoons. It is useful for many different material applications. Versatile aqueous process engineering options can be used to support the morphological and structural modifications of silk materials related to tailored physical, chemical, and biological properties. Conventional solution-based processing methods, while effective, present process control limitations, thus, thermoplastic molding of regenerated SF-based composites was pursued to fabricate dense, functionalized plastics consisting of silk and curcumin. Curcumin, the active compound in turmeric (Curcuma longa) was incorporated into SF during the high-temperature processing, with the objective to investigate composite thermoplastics with enhanced biological properties from the curcumin due to the protective role of silk during processing. The results showed that a significantly higher amount of curcumin (∼25-fold) could be added into thermoplastic molded silk materials compared with the solution route, attributed to the hydrophobicity and low solubility of curcumin in solution-based routes. The curcumin-incorporated silk thermoplastics provided stability in acidic environments like the human gut, and slow curcumin (∼2% over 8 days) release from the materials. The protective silk-curcumin materials supported improved cytocompatibility with immortalized human colorectal adenocarcinoma (Caco-2) cells at high doses. The intestinal epithelial barrier integrity based on zonula occluden 1 (ZO-1) testing showed that the higher amount of curcumin in the thermoplastic molded silk had no negative effects on the intestinal barrier. The functionalized silk-based plastics also displayed microwave stability and antibacterial efficacy against both Gram-positive S. aureus and Gram-negative E. coli. These silk-based sustainable plastics, functionalized with curcumin, offer potential utility for a range of consumer and medical devices.

Enhancing oral bioavailability of dasatinib via supersaturable SNEDDS: Investigation of precipitation inhibition and IVIVC through in-vitro lipolysis-permeation model

Dasatinib (DASA), a potent second-generation multitarget kinase inhibitor marketed as Sprycel® (Tablet), is limited by poor oral bioavailability (14–24 %) and dose-related gastrointestinal side effects. A supersaturable self-nanoemulsifying drug delivery system (su-SNEDDS) designed to enhance DASA’s solubility, sustain supersaturation, and improve oral bioavailability. The su-SNEDDS formulation comprises DASA, an oil, surfactant, co-surfactant, and polyvinylpyrrolidone (PVP) K30 as a precipitation inhibitor (PI). This innovative system demonstrated exceptional stability in gastrointestinal fluids and robustness against dilution, maintaining significantly elevated drug concentrations in the aqueous milieu. su-SNEDDS achieved ∼ 13.5-fold and 2-fold higher aqueous drug concentrations than DASA suspension and SNEDDS without PI, respectively, after 60 min of digestion. This improvement is attributed to the inhibition of crystal growth by PVP K30. In-vitro lipolysis-permeation and Caco-2 cell assays revealed significantly enhanced drug permeation with su-SNEDDS compared to DASA suspension and SNEDDS without PI. In-vivo pharmacokinetic studies further demonstrated ∼ 1.9-fold and 2.7-fold higher AUC compared to SNEDDS without PI and drug suspension, respectively. A linear correlation (R2 = 0.9042) was established between the AUC data obtained from in to vitro vs in-vivo study. These findings underscore the potential of su-SNEDDS to significantly enhance DASA’s solubility, permeation and oral bioavailability, presenting a substantial advancement in pharmaceutical drug delivery systems. Moreover, in-vitro lipolysis-permeation could be promising tool to predict the in-vivo fate of the oral SNEDDS formulations.