Chlorinated paraffins (CPs) are environmental pollutants extensively used in industries. While the use of short-chain chlorinated paraffins (SCCPs) has been restricted since 2017, the use of medium-chain chlorinated paraffins (MCCPs) has risen as their replacement. Due to lipophilic character, it can be expected that CPs enter the cells; however, the in vitro accumulation potential of CPs remains poorly understood. In this study, we aimed to explore the ability of SCCPs and MCCPs to accumulate in fat cells. We utilized an in vitro model of mouse 3T3-L1 preadipocytes and adipocytes. Using gas chromatography coupled with high-resolution mass spectrometry operated in negative chemical ionization mode, we determined the intracellular amounts of CPs. These compounds accumulated at rates of 8.5 ± 0.1 µg/gcells/h for SCCPs and 7.8 ± 0.3 µg/gcells/h for MCCPs when an initial concentration of 120 ng/ml was present in the medium. This rate increased approximately tenfold when the concentration of CPs was raised to 1200 ng/ml. CPs content in adipocytes steadily increased over 5 days, whereas preadipocytes accumulated 15-20 times less CPs. This highlights the importance of cellular lipid content, which was about 12 times higher in adipocytes. Furthermore, we found that the level of chlorine content in the CPs molecules significantly influenced their accumulation. Our results demonstrate that MCCPs exhibit a similar accumulation potential to SCCPs, with lipid content playing a crucial role. As with SCCPs, restrictions on the use of MCCPs in industry should be considered to mitigate their environmental and health impacts.
- Klíčová slova
- 3T3-L1 cells, Adipocytes, Chlorinated paraffins, Persistent organic pollutants,
- MeSH
- buňky 3T3-L1 MeSH
- chlorované uhlovodíky * metabolismus toxicita MeSH
- halogenace MeSH
- látky znečišťující životní prostředí * metabolismus toxicita MeSH
- lipidy analýza MeSH
- metabolismus lipidů * účinky léků MeSH
- myši MeSH
- parafín * metabolismus toxicita chemie MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- tukové buňky * metabolismus účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chlorované uhlovodíky * MeSH
- látky znečišťující životní prostředí * MeSH
- lipidy MeSH
- parafín * MeSH
Cholesterol, the essential building block of cellular membranes, has proven to be a useful tool for increasing the biocompatibility and bioavailability of drug delivery systems in cancer treatment. Resveratrol, a natural polyphenolic compound with promising anticancer properties, faces significant limitations due to its low solubility and bioavailability, hindering its clinical utility. Therefore, in the present study, we aimed to design cholesterol-functionalized cyclodextrin nanosponges (Chol-NSs) with a tunable cholesterol content to optimize resveratrol encapsulation and delivery. Both formulations, free carbonyl diimidazole (CDI) NSs and functionalized Chol-NSs, demonstrated high drug loading and encapsulation efficiency. In vitro experiments revealed that cholesterol incorporation significantly improved the cellular uptake of nanocarriers and potentiated the cytotoxic effects of resveratrol on breast cancer cells. Importantly, both free CDI NSs and functionalized Chol-NSs, even at varying cholesterol percentages, demonstrated a safe profile against both fibroblast and breast cancer cell lines. These results indicate that cholesterol-functionalized nanosponges represent a promising platform for the effective and safe delivery of natural compounds in cancer therapy.
- Klíčová slova
- cholesterol, crosslinked polymer, drug delivery, functionalization, nanosponge,
- MeSH
- biologická dostupnost MeSH
- buňky 3T3 MeSH
- cholesterol * chemie MeSH
- cyklodextriny * chemie MeSH
- imidazoly chemie MeSH
- lékové transportní systémy * metody MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- myši MeSH
- nádory prsu * farmakoterapie patologie MeSH
- nanostruktury * chemie MeSH
- příprava léků metody MeSH
- protinádorové látky aplikace a dávkování farmakokinetika MeSH
- resveratrol * aplikace a dávkování farmakokinetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- cholesterol * MeSH
- cyklodextriny * MeSH
- imidazoly MeSH
- N,N-carbonyldiimidazole MeSH Prohlížeč
- protinádorové látky MeSH
- resveratrol * MeSH
This study deals with utilization of the hyaluronic acid (HA) and carbonyl iron (CI) microparticles to fabricate the magneto-responsive hydrogel scaffolds that can provide triggered functionality upon application of an external magnetic field. The various combinations of the HA and CI were investigated from the rheological and viscoelastic point of view to clearly show promising behavior in connection to 3D printing. Furthermore, the swelling capabilities with water diffusion kinetics were also elucidated. Magneto-responsive performance of bulk hydrogels and their noncytotoxic nature were investigated,, and all hydrogels showed cell viability in the range 75-85%. The 3D printing of such developed systems was successful, and fundamental characterization of the scaffolds morphology (SEM and CT) has been presented. The magnetic activity of the final scaffolds was confirmed at a very low magnetic field strength of 140 kA/m, and such a scaffold also provides very good biocompatibility with NIH/3T3 fibroblasts.
- Klíčová slova
- 3D printing, hyaluronic acid, magnetic particles, magneto-responsive, scaffold,
- MeSH
- 3D tisk * MeSH
- biokompatibilní materiály * chemie farmakologie MeSH
- biopolymery chemie MeSH
- buňky NIH 3T3 MeSH
- hydrogely chemie farmakologie MeSH
- karbonylové sloučeniny železa chemie MeSH
- kyselina hyaluronová * chemie farmakologie MeSH
- myši MeSH
- testování materiálů * MeSH
- tkáňové podpůrné struktury * chemie MeSH
- velikost částic * MeSH
- viabilita buněk * účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biokompatibilní materiály * MeSH
- biopolymery MeSH
- hydrogely MeSH
- karbonylové sloučeniny železa MeSH
- kyselina hyaluronová * MeSH
The molecular mechanisms linking obstructive sleep apnea syndrome (OSA) to obesity and the development of metabolic diseases are still poorly understood. The role of hypoxia (a characteristic feature of OSA) in excessive fat accumulation has been proposed. The present study investigated the possible effects of hypoxia (4% oxygen) on de novo lipogenesis by tracking the major carbon sources in differentiating 3T3-L1 adipocytes. Gas-permeable cultuware was employed to cultivate 3T3-L1 adipocytes in hypoxia (4%) for 7 or 14 days of differentiation. We investigated the contribution of glutamine, glucose or acetate using 13C or 14C labelled carbons to the newly synthesized lipid pool, changes in intracellular lipid content after inhibiting citrate- or acetate-dependent pathways and gene expression of involved key enzymes. The results demonstrate that, in differentiating adipocytes, hypoxia decreased the synthesis of lipids from glucose (44.1 ± 8.8 to 27.5 ± 3.0 pmol/mg of protein, p < 0.01) and partially decreased the contribution of glutamine metabolized through the reverse tricarboxylic acid cycle (4.6% ± 0.2-4.2% ± 0.1%, p < 0.01). Conversely, the contribution of acetate, a citrate- and mitochondria-independent source of carbons, increased upon hypoxia (356.5 ± 71.4 to 649.8 ± 117.5 pmol/mg of protein, p < 0.01). Further, inhibiting the citrate- or acetate-dependent pathways decreased the intracellular lipid content by 58% and 73%, respectively (p < 0.01) showing the importance of de novo lipogenesis in hypoxia-exposed adipocytes. Altogether, hypoxia modified the utilization of carbon sources, leading to alterations in de novo lipogenesis in differentiating adipocytes and increased intracellular lipid content.
- MeSH
- acetáty * metabolismus farmakologie MeSH
- buněčná diferenciace * účinky léků MeSH
- buňky 3T3-L1 * MeSH
- citrátový cyklus MeSH
- glukosa * metabolismus MeSH
- glutamin * metabolismus MeSH
- hypoxie buňky MeSH
- lipidy biosyntéza MeSH
- lipogeneze * účinky léků MeSH
- metabolismus lipidů účinky léků MeSH
- myši MeSH
- tukové buňky * metabolismus účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- acetáty * MeSH
- glukosa * MeSH
- glutamin * MeSH
- lipidy MeSH
The biologically produced gold nanoparticles (AuNPs) are novel carriers with promising use in targeted tumor therapy. Still, there are no studies regarding the efficacy of nanoparticle internalization by cancer and noncancer cells. In this study, AuNPs were produced by Fusarium oxysporum and analyzed by spectrophotometry, transmission electron microscopy (TEM), energy dispersive x-ray spectroscopy (EDS), and Zetasizer. Obtained AuNPs were about 15 nm in size with a zeta potential of -35.8 mV. The AuNPs were added to cancer cells (4T1), noncancer cells (NIH/3T3), and macrophages (RAW264.7). The viability decreased in 4T1 (77 ± 3.74%) in contrast to NIH/3T3 and RAW264.7 cells (89 ± 4.9% and 90 ± 3.5%, respectively). The 4T1 cancer cells also showed the highest uptake and accumulation of Au (∼80% of AuNPs was internalized) as determined by graphite furnace atomic absorption spectroscopy. The lowest amount of AuNPs was internalized by the NIH/3T3 cells (∼30%). The NIH/3T3 cells exhibited prominent reorganization of F-actin filaments as examined by confocal microscopy. In RAW264.7, we analyzed the release of proinflammatory cytokines by flow cytometry and we found the AuNP interaction triggered transient secretion of tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ). In summary, we proved the biologically produced AuNPs entered all the tested cell types and triggered cell-specific responses. High AuNP uptake by tumor cells was related to decreased cell viability, while low nanoparticle uptake by fibroblasts triggered F-actin reorganization without remarkable toxicity. Thus, the biologically produced AuNPs hold promising potential as cancer drug carriers and likely require proper surface functionalization to shield phagocytizing cells.
- Klíčová slova
- 4T1 cells, NIH/3T3 cells, RAW264.7 cells, actin reorganization, biologically produced gold nanoparticles, nanoparticle uptake, stress response,
- MeSH
- buňky NIH 3T3 MeSH
- Fusarium metabolismus MeSH
- kovové nanočástice * chemie MeSH
- makrofágy metabolismus účinky léků MeSH
- myši MeSH
- RAW 264.7 buňky MeSH
- viabilita buněk účinky léků MeSH
- zlato * chemie metabolismus farmakologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- zlato * MeSH
BACKGROUND: Obesity is a major health burden. Preadipocytes proliferate and differentiate in mature adipocytes in the adipogenic process, which could be a potential therapeutic approach for obesity. Deficiency of SIRT6, a stress-responsive protein deacetylase and mono-ADP ribosyltransferase enzyme, blocks adipogenesis. Mutants of SIRT6 (N308K/A313S) were recently linked to the in the long lifespan Ashkenazi Jews. In this study, we aimed to clarify how these new centenarian-associated SIRT6 genetic variants affect adipogenesis at the transcriptional and epigenetic level. METHODS: We analyzed the role of SIRT6 wild-type (WT) or SIRT6 centenarian-associated mutant (N308K/A313S) overexpression in adipogenesis, by creating stably transduced preadipocyte cell lines using lentivirus on the 3T3-L1 model. Histone post-translational modifications (PTM: acetylation, methylation) and transcriptomic changes were analyzed by mass spectrometry (LC-MS/MS) and RNA-Seq, respectively, in 3T3-L1 adipocytes. In addition, the adipogenic process and related signaling pathways were investigated by bioinformatics and biochemical approaches. RESULTS: Overexpression of centenarian-associated SIRT6 mutant increased adipogenic differentiation to a similar extent compared to the WT form. However, it triggered distinct histone PTM profiles in mature adipocytes, with significantly higher acetylation levels, and activated divergent transcriptional programs, including those dependent on signaling related to the sympathetic innervation and to PI3K pathway. 3T3-L1 mature adipocytes overexpressing SIRT6 N308K/A313S displayed increased insulin sensitivity in a neuropeptide Y (NPY)-dependent manner. CONCLUSIONS: SIRT6 N308K/A313S overexpression in mature adipocytes ameliorated glucose sensitivity and impacted sympathetic innervation signaling. These findings highlight the importance of targeting SIRT6 enzymatic activities to regulate the co-morbidities associated with obesity.
- Klíčová slova
- Adipogenesis, Epigenetics, Histones, Obesity, SIRT6,
- MeSH
- adipogeneze * genetika MeSH
- buňky 3T3-L1 * MeSH
- epigeneze genetická * genetika MeSH
- histony metabolismus genetika MeSH
- lidé MeSH
- mutace MeSH
- myši MeSH
- obezita genetika metabolismus MeSH
- posttranslační úpravy proteinů genetika MeSH
- sirtuiny * genetika metabolismus MeSH
- tukové buňky * metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- histony MeSH
- SIRT6 protein, human MeSH Prohlížeč
- Sirt6 protein, mouse MeSH Prohlížeč
- sirtuiny * MeSH
BACKGROUND: Obesity and related metabolic diseases are becoming a worldwide epidemic, leading to increased mortality and heavy medical costs. Our Chinese herbal formula Xiao-Gao-Jiang-Zhuo (XGJZ) has remarkable effects on curing obese patients in the clinic, but the cellular and molecular basis remains unknown. This study aimed to reveal the molecular mechanism involved in adipogenesis in vitro. METHODS: Chinese herbal formula XGJZ-containing serum was prepared from XGJZ-treated obesity model rats. The function of XGJZ-containing serum was validated in 3T3-L1 preadipocytes. Oil O staining was performed to determine intracellular lipid accumulation in differentiated 3T3-L1 cells. The expression of pro-adipogenic transcription factors was measured to further validate the adipogenesis of 3T3-L1 adipocytes. The contents of triglyceride (TG), free fatty acid (FFA), and glycerin, along with the activities of lipid metabolism-related enzymes (including FAT, FATP1, DGAT, GPAT, ATGL, and HSL) were measured to study the lipogenesis in 3T3-L1 adipocytes. RESULTS: XGJZ-containing serum inhibited 3T3-L1 differentiation, decreased intracellular lipid accumulation, and suppressed the expression of pro-adipogenic transcription factors in differentiated 3T3-L1 cells. The contents of TG, FFA, and glycerin were decreased when treated with XGJZ-containing serum, which also modulated lipid metabolism-related enzyme activities. The activities of fatty acid transporters (FAT, FATP1) and lipid mobilization enzymes (ATGL, HSL) were promoted, while activities of triglyceride biosynthesis enzymes (DGAT, GPAT) were attenuated in differentiated 3T3-L1 cells. CONCLUSION: XGJZ-containing serum has inhibitory effects on adipogenesis in 3T3-L1 preadipocytes, affirming the effect of XGJZ in treating obesity. It provides evidence for the mechanism of obesity.
- Klíčová slova
- 3T3-L1 preadipocytes, XGJZ, adipokines, lipid accumulation,
- MeSH
- adipogeneze * MeSH
- buňky 3T3-L1 MeSH
- glycerol * metabolismus farmakologie MeSH
- krysa rodu Rattus MeSH
- lidé MeSH
- myši MeSH
- obezita MeSH
- PPAR gama metabolismus MeSH
- transkripční faktory MeSH
- triglyceridy MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- glycerol * MeSH
- PPAR gama MeSH
- transkripční faktory MeSH
- triglyceridy MeSH
Major challenges facing clinicians treating burn wounds are the lack of integration of treatment to wound, inadequate mechanical properties of treatments, and high infection rates which ultimately lead to poor wound resolution. Electrospun chitosan membranes (ESCM) are gaining popularity for use in tissue engineering applications due to their drug loading ability, biocompatibility, biomimetic fibrous structure, and antimicrobial characteristics. This work aims to modify ESCMs for improved performance in burn wound applications by incorporating elastin and magnesium-phosphate particles (MgP) to improve mechanical and bioactive properties. The following ESCMs were made to evaluate the individual components' effects; (C: chitosan, CE: chitosan-elastin, CMg: chitosan-MgP, and CEMg: chitosan-elastin-MgP). Membrane properties analyzed were fiber size and structure, hydrophilic properties, elastin incorporation, MgP incorporation and in vitro release, mechanical properties, degradation profiles, and in vitro cytocompatibility with NIH3T3 fibroblasts. The addition of both elastin and MgP increased the average fiber diameter of CE (~400 nm), CMg (~360 nm), and CEMg (565 nm) compared to C (255 nm). Water contact angle analysis showed elastin incorporated membranes (CE and CEMg) had increased hydrophilicity (~50°) compared to the other groups (C and CMg, ~110°). The results from the degradation study showed mass retention of ~50% for C and CMg groups, compared to ~ 30% seen in CE and CEMg after 4 weeks in a lysozyme/PBS solution. CMg and CEMg exhibited burst-release behavior of ~6 µg/ml or 0.25 mM magnesium within 72 h. In vitro analysis with NIH3T3 fibroblasts showed CE and CEMg groups had superior cytocompatibility compared to C and CMg. This work has demonstrated the successful incorporation of elastin and MgP into ESCMs and allows for future studies on burn wound applications.
- Klíčová slova
- chitosan, elastin, electrospinning, tissue engineering, wound healing,
- MeSH
- antiinfekční látky * farmakologie MeSH
- buňky NIH 3T3 MeSH
- chitosan * chemie MeSH
- elastin MeSH
- fosfáty MeSH
- hojení ran MeSH
- hořčík MeSH
- muramidasa farmakologie MeSH
- myši MeSH
- nanovlákna * chemie MeSH
- popálení * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antiinfekční látky * MeSH
- chitosan * MeSH
- elastin MeSH
- fosfáty MeSH
- hořčík MeSH
- muramidasa MeSH
Clinically approved photodynamic therapy (PDT) is a minimally invasive treatment procedure that uses three key components: photosensitization, a light source, and tissue oxygen. However, the photodynamic effect is limited by both the photophysical properties of photosensitizers as well as their low selectivity, leading to damage to adjacent normal tissue and/or inadequate biodistribution. Nanoparticles (NPs) represent a new option for PDT that can overcome most of the limitations of conventional photosensitizers and can also promote photosensitizer accumulation in target cells through enhanced permeation and retention effects. In this in vitro study, the photodynamic effect of TPP photosensitizers embedded in polystyrene nanoparticles was observed on the non-tumor NIH3T3 cell line and HeLa and G361 tumor cell lines. The efficacy was evaluated by viability assay, while reactive oxygen species production, changes in membrane mitochondrial potential, and morphological changes before and after treatment were imaged by atomic force microscopy. The tested nanoparticles with embedded TPP were found to become cytotoxic only after activation by blue light (414 nm) due to the production of reactive oxygen species. The photodynamic effect observed in this evaluation was significantly higher in both tumor lines than the effect observed in the non-tumor line, and the resulting phototoxicity depended on the concentration of photosensitizer and irradiation time.
- Klíčová slova
- cancer, nanoparticles, photodynamic effect,
- MeSH
- buňky NIH 3T3 MeSH
- fotochemoterapie * metody MeSH
- fotosenzibilizující látky farmakologie terapeutické užití MeSH
- lidé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nanočástice * MeSH
- porfyriny * metabolismus farmakologie MeSH
- reaktivní formy kyslíku metabolismus MeSH
- tkáňová distribuce MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- fotosenzibilizující látky MeSH
- porfyriny * MeSH
- reaktivní formy kyslíku MeSH
A little is known about the link between the macromolecular architecture of dialdehyde polysaccharides (DAPs), their crosslinking capabilities, and the properties of resulting hydrogels. Here, DAPs based on cellulose, dextrin, dextran, and hyaluronate were compared as crosslinkers for poly(vinyl alcohol), PVA. The swelling, network parameters, viscoelastic properties, porosity, and cytotoxicity of PVA/DAP hydrogels were investigated concerning the crosslinker structure, molecular weight, aldehyde group density per macromolecule, and the size of spontaneously formed crosslinker nano-assemblies. Generally, crosslinkers based on linear polysaccharides (cellulose, hyaluronate) performed more reliably, while the presence of branching could be both beneficial (dextran) but also detrimental (dextrin) at lower crosslinker concentrations. For example, the hydrogel swelling differed by up to one-third (600 vs. 400%) and storage modulus even by up to one half (~7000 vs. ~3500 Pa) depending on crosslinker structure and properties. These differences were rationalized by variances in crosslinking modes derived based on obtained data.
- Klíčová slova
- Cellulose, Crosslinking, Dextran, Dextrin, Dialdehyde, Hyaluronic acid, Poly(vinyl) alcohol,
- MeSH
- buňky NIH 3T3 MeSH
- hydrogely chemie farmakologie MeSH
- myši MeSH
- polysacharidy chemie farmakologie MeSH
- polyvinylalkohol chemie farmakologie MeSH
- reagencia zkříženě vázaná chemie farmakologie MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- hydrogely MeSH
- polysacharidy MeSH
- polyvinylalkohol MeSH
- reagencia zkříženě vázaná MeSH
