Proliferation Of Cultured Cells Research Articles

The Antidepressant Drug Amitriptyline Affects Human SH-SY5Y Neuroblastoma Cell Proliferation and Modulates Autophagy.

Amitriptyline is a tricyclic antidepressant commonly used for depressive disorders and is prescribed off-label for several neurological conditions like neuropathic pain, migraines and anxiety. Besides their action on the reuptake of monoaminergic neurotransmitters, tricyclic antidepressants interact with several additional targets that may contribute to either therapeutic or adverse effects. Here, we investigated the effects of amitriptyline on proliferation and autophagy (i.e., an evolutionarily conserved catabolic pathway responsible for the degradation and recycling of cytoplasmic material) in human SH-SY5Y neuroblastoma cell cultures. The dose and time-dependent upregulation of the autophagy marker LC3II and the autophagy receptor p62, with the accumulation of LAMP1 positive compartments, were observed in SH-SY5Y cells exposed to the amitriptyline. These effects were accompanied by reduced cell viability and decreased clonogenic capacity, without a significant induction of apoptosis. Decrease viability and clonogenic activity were still observed in autophagy deficient Atg5-/- MEF and following pre-treatment of SH-SY5Y culture with the autophagy inhibitor chloroquine, suggesting that they were independent from autophagy modulation. Our findings demonstrate that amitriptyline acts on pathways crucial for cell and tissue homeostasis (i.e., autophagy and proliferation) and pose the basis for further studies on the potential therapeutic application of amitriptyline, as well as the consequences of its use for long-term treatments.

Morphological, cytogenetic, and phytochemical estimation of Lychnis flos-cuculi L. cell cultures: can cell biomass be a source of bioactive compounds?

The ragged robin (Lychnis flos-cuculi L.) from the Caryophyllaceae family contains flavonoids, phenolic acids, flavonoids, triterpenoid saponins, and ecdysteroids, making it potential for biological and medicinal application. Plant cell culture technologies could be beneficial for obtaining low-cost biomass sources with valuable phytochemicals sustainably. The current study reports the induction and proliferation of ragged robin cell culture and evaluation of various plant growth regulators (PGRs) impact on the secondary metabolites profile using UPLC-HRMS/MS. Callus was initiated from different explants on the Murashige and Skoog medium with various combinations and concentrations of PGRs. It was found that prolonged proliferation on MS medium enriched with a combination of 1.0 mg l− 1 2,4-Dichlorophenoxyacetic acid and 0.1 mg l− 1 N6-furfuryl adenine (Kinetin) yields the maximum biomass with 504.05% growth index. The results of flow cytometric analysis suggested no genetic alteration between differentiated and dedifferentiated cultures. Furthermore, the UPLC-HRMS/MS result distinguished 101 significant signals that were manually identified as ecdysteroids (16 metabolites), flavonoids (11 metabolites), phenolic acids (10 metabolites), triterpene saponins and sapogenins (53 metabolites), and some primary metabolites. Our study demonstrates that the cell culture of L. flos-cuculi displays significant differences in its phytochemical makeup to plants cultivated in vitro. Moreover, the type of growth regulators used for callus cultures was identified to cause substantial shifts in the identified metabolites.Graphical

Cell Culture Differentiation and Proliferation Conditions Influence the InVitro Regeneration of the Human Airway Epithelium.

The human airway mucociliary epithelium can be recapitulated invitro using primary cells cultured in an air-liquid interface (ALI), a reliable surrogate to perform pathophysiological studies. As tremendous variations exist among media used for ALI-cultured human airway epithelial cells, the aim of our study was to evaluate the impact of several media (BEGM, PneumaCult, Half & Half, and Clancy) on cell type distribution using single-cell RNA sequencing and imaging. Our work revealed the impact of these media on cell composition, gene expression profile, cell signaling, and epithelial morphology. We found higher proportions of multiciliated cells in PneumaCult-ALI and Half & Half, stronger EGF signaling from basal cells in BEGM-ALI, differential expression of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) entry factor ACE2, and distinct secretome transcripts depending on the media used. We also established that proliferation in PneumaCult-Ex Plus favored secretory cell fate, showing the key influence of proliferation media on late differentiation epithelial characteristics. Altogether, our data offer a comprehensive repertoire for evaluating the effects of culture conditions on airway epithelial differentiation and will aid in choosing the most relevant medium according to the processes to be investigated, such as cilia, mucus biology, or viral infection. We detail useful parameters that should be explored to document airway epithelial cell fate and morphology.

Study on the Effects of Low-Intensity Pulsed Ultrasound and Iron Ions for Proliferation and Differentiation of Osteoblasts

ObjectiveThis study involved the proliferation and differentiation of osteoblasts treated with low-intensity pulsed ultrasound (LIPUS) and iron (Fe3+) ions, respectively. The biological effects of LIPUS and Fe3+ ions on the proliferation and differentiation of osteoblasts were also evaluated. MethodsMC3T3-E1 cells were seeded in six-well plates with the medium, which contained different concentrations of Fe3+ (0, 100, 200, 300, 400, 500, 600 and 700 μg L-1, respectively). LIPUS treatment was directed at the bottom of the plate for 20 min at an intensity of 80 mW cm-2 every day. ResultsViability results showed that a dose of 400 μg L-1 Fe3+ ions had the best effect at promoting osteogenic proliferation in cell culture. The results of alkaline phosphatase staining and mineralization indicated that the differentiation of osteoblasts was promoted by LIPUS and Fe3+ ions. Fluorescence staining results showed that the number of cell nuclei in the LIPUS, Fe3+ and LIPUS-Fe groups increased by 37.20%, 55.81% and 89.76%, respectively. Migration data indicated that migration and proliferation rates were increased by LIPUS and Fe3+, and the results of protein expression indicated that LIPUS and Fe3+ may increase the expression of Wnt, β-catenin, and Runx2, hence promoting normal bone regeneration and development. ConclusionThe combination of LIPUS (1.5 MHz, 80 mW cm-2) and Fe3+ accelerates the proliferation and differentiation of osteoblasts significantly compared with single-factor treatment (stimulated by LIPUS and Fe3+ ions, respectively). This study could establish a foundation for LIPUS-responsive biomaterials in the repair and regeneration of bone tissues.

Microencapsulation of betacyanins accumulated calli, short-term cold storage, and retrieval of callus in Selenicereus undatus (Haw.) D.R. Hunt

The red color of dragon fruit pulp and peel is due to the accumulation of betacyanins. In the present study, callus cultures were established using in vitro germinated seedlings of the white dragon fruit plant (Selenicereus undatus). Murashige and Skoog (MS) medium with 6-benzylaminopurine (BAP) at 2.0 mg/L was used to induce red-colored friable callus (92.0 % response). The betacyanin accumulated calli were harvested and encapsulated using sodium alginate (NaC6H7O6) and calcium chloride (CaCl2). These synseeds were cold stored for 6 months at 4.0 °C temperature. The stored synseeds were germinated on 2.0 mg/L BAP in the medium. The highest germination rate (88 %) of synseeds was achieved with 3 % sodium alginate polymerized with 75 mM calcium chloride, resulting in firm, uniform, and round synseeds. The betacyanin-loaded synseeds could be stored, transported, and directly used as inoculums for the extensive proliferation of calli and cell cultures for the commercial production of betacyanin.

Cytotoxicity of Polymer Scaffolds Suitable for Manufacturing of Small-Diameter Vascular Grafts

Aim.To evaluate the cytotoxicity of poly(ε-caprolactone) and polyurethane scaffolds in vitro.Materials and Methods. Polymer scaffolds were made by electrospinning from a 12% solution of poly(ε-caprolactone) or a 12% solution of polyurethane. Surface structure was examined by scanning electron microscopy, whilst cytotoxicity was evaluated by seeding EA.hy 926 endothelial cells on scaffold surface for 72 hours. Cell culture viability and proliferation was assessed by MTT assay and by quantifying cell culture density. On the xCELLigence device, cells were cultured in the presence of the studied matrix samples, and the dynamics of cell culture growth was evaluated in real time.Results. Poly(ε-caprolactone) scaffolds were characterised by a higher variability in the filament thickness and by a significantly larger pore size. Polyurethane filaments formed a dense web with a smoother surface. Poly(ε-caprolactone) scaffolds had significantly higher biocompatibility in comparison with polyurethane. Adhesion of cells to poly(ε-caprolactone) scaffolds did not differ from the cell culture plastic, and poly(ε-caprolactone) supported cell proliferation in the MTT test. Poly(ε-caprolactone) and polyurethane did not differ significantly in terms of inducing cell proliferation. Both poly(ε-caprolactone) and polyurethane scaffolds did not pose considerable cytotoxicity.Conclusion. Poly(ε-caprolactone) and polyurethane scaffolds did not exhibit cytotoxic effects and can be used for manufacturing polymer scaffolds of vascular grafts.

A MOZ-TIF2 leukemia mouse model displays KAT6-dependent H3K23 propionylation and overexpression of a set of active developmental genes

Aberrant regulation of chromatin modifiers is a common occurrence across many cancer types, and a key priority is to determine how specific alterations of these proteins, often enzymes, can be targeted therapeutically. MOZ, a histone acyltransferase, is recurrently fused to coactivators CBP, p300, and TIF2 in cases of acute myeloid leukemia (AML). Using either pharmacological inhibition or targeted protein degradation in a mouse model for MOZ-TIF2-driven leukemia, we show that KAT6 (MOZ/MORF) enzymatic activity and the MOZ-TIF2 protein are necessary for indefinite proliferation in cell culture. MOZ-TIF2 directly regulates a small subset of genes encoding developmental transcription factors, augmenting their high expression. Furthermore, transcription levels in MOZ-TIF2 cells positively correlate with enrichment of histone H3 propionylation at lysine 23 (H3K23pr), a recently appreciated histone acylation associated with gene activation. Unexpectedly, we also show that MOZ-TIF2 and MLL-AF9 regulate transcription of unique gene sets, and their cellular models exhibit distinct sensitivities to multiple small-molecule inhibitors directed against AML pathways. This is despite the shared genetic pathways of wild-type MOZ and MLL. Overall, our data provide insight into how aberrant regulation of MOZ contributes to leukemogenesis. We anticipate that these experiments will inform future work identifying targeted therapies in the treatment of AML and other diseases involving MOZ-induced transcriptional dysregulation.

A low dose of curcumin-PDA nanoparticles improves viability and proliferation in endoneurial fibroblasts and Schwann cell cultures

Curcumin is a polyphenol extracted from Curcuma longa’s roots. Low doses of curcumin are related to anti-inflammatory, antioxidant, and neuroprotective effects, while high doses are used for their lethality. This diversity of behaviors allows us to understand curcumin as a compound with hormetic action. Due to its strongly hydrophobic character, curcumin is often solubilized in organic compounds. In this way, we have recently reported the undesirable and occasionally irreversible effects of alcohol and DMSO on the viability of primary Schwann cell cultures. In this scenario, the use of nanoparticles as delivery systems has become a successful alternative strategy for these compounds. In the present work, we describe the structure of Polydopamine (PDA) nanoparticles, loaded with a low dose of curcumin (Curc-PDA) without the use of additional organic solvents. We analyzed the curcumin released, and we found two different forms of curcumin. Small increased cell viability and proliferation were observed in endoneurial fibroblast and Schwann cell primary cultures when Curc-PDA was steadily supplied for 5 days. The increased bioavailability of this natural compound and the impact on cells in culture not only confirm the properties of curcumin at very low doses but also provide a glimpse of a possible therapeutic alternative for PNS conditions in which SCs are involved.

Targeting CXCR4/CXCL12 axis via [177Lu]Lu-DOTAGA.(SA.FAPi)2 with CXCR4 antagonist in triple-negative breast cancer

PurposeRadiopharmaceutical therapies targeting fibroblast activation protein (FAP) have shown promising efficacy against many tumor types. But radiopharmaceuticals alone in most cases are insufficient to completely eradicate tumor cells, which can partially be attributed to the protective interplay between tumor cells and cancer-associated fibroblasts (CAFs). The C-X-C chemokine receptor type 4/C-X-C motif chemokine 12 (CXCR4/CXCL12) interaction plays an important role in orchestrating tumor cells and CAFs. We hereby investigated the feasibility and efficacy of [177Lu]Lu-DOTAGA.(SA.FAPi)2, a FAP-targeting radiopharmaceutical, in combination with AMD3100, a CXCR4 antagonist, in a preclinical murine model of triple-negative breast cancer (TNBC).MethodsPublic database was first interrogated to reveal the correlation between CAFs’ scores and the prognosis of TNBC patients, as well as the expression levels of FAP and CXCR4 in normal tissues and tumors. In vitro therapeutic efficacy regarding cell proliferation, migration, and colony formation was assessed in BALB/3T3 fibroblasts and 4T1 murine breast cancer cells. In vivo therapeutic efficacy was longitudinally monitored using serial 18F-FDG, [18F]AlF-NOTA-FAPI-04, and [68Ga]Ga-DOTA-Pentixafor PET/CT scans and validated using tumor sections through immunohistochemical staining of Ki-67, α-SMA, CXCR4, and CXCL12. Intratumoral abundance of myeloid-derived suppressive cells (MDSCs) was analyzed using flow cytometry in accordance with the PET/CT schedules. Treatment toxicity was evaluated by examining major organs including heart, lung, liver, kidney, and spleen.ResultsCAFs’ scores negatively correlated with the survival of TNBC patients (p < 0.05). The expression of CXCR4 and FAP was both significantly higher in tumors than in normal tissues. The combination of [177Lu]Lu-DOTAGA.(SA.FAPi)2 and AMD3100 significantly suppressed cell proliferation, migration, and colony formation in cell culture, and exhibited synergistic effects in 4T1 tumor models along with a decreased number of MDSCs. PET/CT imaging revealed lowest tumor accumulation of 18F-FDG and [18F]AlF-NOTA-FAPI-04 on day 13 and day 14 after treatment started, both of which gradually increased at later time points. A similar trend was observed in the IHC staining of Ki-67, α-SMA, and CXCL12.ConclusionThe combination of [177Lu]Lu-DOTAGA.(SA.FAPi)2 and AMD3100 is a feasible treatment against TNBC with minimal toxicity in main organs.

The toxigenic red-tide-forming dinoflagellates Alexandrium leei and Alexandrium catenella differ in terms of the sensitivity to strong light and low temperature of their photosynthetic machinery

Alexandrium leei and Alexandrium catenella are noxious dinoflagellates that form red tides but the two species differ substantially in optimal growth conditions. For formation of blooms in natural environments, A. leei favors moderate temperatures (around 20 °C) while A. catenella favors much lower temperatures (around 10 °C). We examined the effects of temperature and light on cell proliferation and photosynthesis in cell cultures of the two species. Under light at 100 μmol photons m−2 s−1, the growth rate of both species was highest at 20 °C. At 25 °C, A. leei continued to proliferate but A. catenella did not survive. Conversely, at lower temperatures, namely, 12 °C and 15 °C, the growth rate of A. leei was very low while that of A. catenella remained high. The net photosynthetic activity, as determined in terms of oxygen evolution, was highest at 20 °C in A. leei. By contrast, it was highest at 15 °C in A. catenella. Illumination of cells with strong light at 1000 μmol photons m−2 s−1 decreased the activity of photosystem II (PSII), as determined in terms of Fv/Fm, in both species. In A. leei, the activity of PSII declined more rapidly at 12 °C and 15 °C than at 20 °C and 25 °C under strong light. In A. catenella, the activity of PSII declined more rapidly at 20 °C than at 12 °C and 15 °C. Thus, A. catenella appeared to be more resistant to photoinhibition of PSII at low temperatures than A. leei. In addition, the ability to repair photodamaged PSII at 12 °C and 15 °C was greater in A. catenella than that in A. leei. Thus, the tolerance of PSII to strong light and low temperatures might determine the range of temperatures that supports the formation of blooms of the two Alexandrium species.

Nonwoven fiber meshes for oxygen sensing

Accurate oxygen sensing and cost-effective fabrication are crucial for the adoption of wearable devices inside and outside the clinical setting. Here we introduce a simple strategy to create nonwoven polymeric fibrous mats for a notable contribution towards addressing this need. Although morphological manipulation of polymers for cell culture proliferation is commonplace, especially in the field of regenerative medicine, non-woven structures have not been used for oxygen sensing. We used an airbrush spraying, i.e. solution blowing, to obtain nonwoven fiber meshes embedded with a phosphorescent dye. The fibers serve as a polymer host for the phosphorescent dye and are shown to be non-cytotoxic. Different composite fibrous meshes were prepared and favorable mechanical and oxygen-sensing properties were demonstrated. A Young's modulus of 9.8 MPa was achieved and the maximum oxygen sensitivity improved by a factor of ∼2.9 compared to simple drop cast film. The fibers were also coated with silicone rubbers to produce mechanically robust sensing films. This reduced the sensing performance but improved flexibility and mechanical properties. Lastly, we are able to capture oxygen concentration maps via colorimetry using a smartphone camera, which should offer unique advantages in wider usage. Overall, the introduced composite fiber meshes show a potential to significantly improve cell cultures and healthcare monitoring via absolute oxygen sensing.

Abstract 4387: Interplay of Notch1 and Notch3 signaling regulates pancreatic neuroendocrine tumor proliferation

Abstract Background: The 5-year survival rate for patients with advanced pancreatic neuroendocrine tumors (pNETs) is less than 30%. Notch signaling is a transmembrane receptor pathway with four receptor isoforms and five different ligands. Binding between a receptor and ligand triggers pathway activation and is linked to cellular differentiation, cell fate, proliferation, and viability. Both Notch1 and Notch3 signaling have been shown to be dysregulated in pNETs. Notch3 activation has been shown to repress Notch1 activity in other cancers. We hypothesize that activation of Notch3 can reduce Notch1-mediated proliferation in pNETs. Methods: Genetic deletion of Notch1 (N1-KO) in a pNET cell line (BON) was achieved using CRISPR/Cas9 to create a compound heterozygous knockout at exon 3. An inducible Notch1 overexpression cell line was generated by stably transfecting BON with a plasmid construct containing the Notch1 active form (N1ICD) under a Tet-On regulator. A constitutively active Notch3 cell line was generated by lentiviral transduction of BON with a plasmid overexpressing the Notch3 active form (N3ICD). Real-time quantitative PCR was used to measure mRNA transcript levels, and western blot was used to measure protein expression. Two-dimensional (2D) viability was measured by MTT assay, and three-dimensional (3D) proliferation was measured by spheroid size. Results: Genetic knockout of Notch1 (N1-KO) reduced the expression of Notch3 at both mRNA and protein levels. N1-KO had reduced proliferation compared to (WT) BON. Increased cell density (ligand availability) or treatment with valproic acid (known Notch1 inducer) led to increased N3ICD in WT BON but not in N1-KO. Overexpression of N1ICD was associated with increased expression of a known downstream target Hes1, as well as Notch3. In contrast, forced overexpression of N3ICD led to increased Notch1 expression, but not Hes1 when compared to a BON cell line transfected with an empty vector. Functionally, N1-KO and N3ICD overexpression cells both had reduced proliferation in both 2D and 3D cell culture. Conclusions: The Notch1 and Notch3 receptors produce signaling cascades that regulate growth in pNET cells through distinct mechanisms. Reduction of Notch1 signaling suppresses growth, while the same effect can be achieved with Notch3 overexpression. Thus, specifically upregulating Notch3 may be a strategy to block Notch1-mediated proliferation in pNET cells. Citation Format: Rachael Guenter, Weisheng Chen, Yuvasri Golivi, Mario Robledo, Cole Adams, Renata Jaskula-Sztul, Herbert Chen, J. Bart Rose. Interplay of Notch1 and Notch3 signaling regulates pancreatic neuroendocrine tumor proliferation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4387.

Abstract 1631: A homogeneous assay system for discernment of proliferative, antiproliferative, and cytotoxic effects in culture

Abstract We describe a novel homogeneous cell-based assay developed for the quantitative detection of Ki-67 protein, a recognized marker of cell proliferation, utilizing Lumit™ immunoassay technology. This luminescent assay has been effectively multiplexed with a fluorescent measure of cell death, employing a cell-impermeant fluorogenic DNA dye. This multiplex allows for the clear distinction between antiproliferative activity and overt cytotoxicity in cell culture models. Through varying lytic conditions, we demonstrated effective cell lysis and detection of nuclear Ki-67 protein. Specifically, Lumit™ Lysis Buffer II provided efficient recovery of Ki-67 protein without compromising assay performance, including the requisite binding of labeled antibodies to the target protein. This assay exhibited excellent signal-to-background (up to 40-fold) and signal linearity (R^2 = 0.99) in wildtype HeLa cells (~1000 to 80,000/well), while the signal in Ki-67 knockout HeLa cells was comparable to the no-cell background, indicating excellent specificity for Ki-67 protein. Due to the unavailability of recombinant full-length Ki-67 protein, two different recombinant Ki-67 protein fragments were used to corroborate assay sensitivity of &amp;lt;1 pM Ki-67 protein. The assay's suitability for human Ki-67 protein detection, not mouse, was observed through further cell-based testing. The proliferation/cytotoxicity assay multiplex was validated in both suspension and adherent cell lines, as well as primary cell culture, detecting antiproliferative and proliferative responses. CDK4/6 inhibitors are a class of antiproliferative agents of significant interest for their potential in treating cancer. In Jurkat cells, treatment with the CDK4/6 inhibitor palbociclib led to a dose-dependent reduction in Ki-67 protein expression with an EC50 of 88 nM, and total Ki-67 elimination at 2 μM compound concentration. Notably, there was no cytotoxicity detected at the tested doses and treatment duration (up to 48 h), but a significant reduction in viable cell number at maximal palbociclib concentrations, aligning with an antiproliferative mechanism. This work establishes a rapid, simple, luminescent assay for determining Ki-67 levels that can be paired with a fluorogenic readout of cell death, facilitating efficient and quantitative discernment of altered proliferation in mammalian cell culture using standard microplate readers. Citation Format: Dan F. Lazar, Kevin R. Kupcho, Andrew L. Niles, James J. Cali. A homogeneous assay system for discernment of proliferative, antiproliferative, and cytotoxic effects in culture [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1631.

Thioredoxin 2 protects mice against experimental myocardial infarction

Graphical abstract Experimental Myocardial infarction (MI) using ligation procedure induces cardiac dysfunction, high level of ROS, inflammation, apoptosis, fibrosis and cardiomyocyte (CM) loss. AAV overexpressing human Trx-2, specifically in CM mitochondria improves mouse cardiac function, reduces the size of cardiac infarct, increases the expression of cardiac anti-inflammatory markers, reduces apoptosis and oxidative stress. However, it does not increase CM proliferation. Abstract Introduction and objective Myocardial infarction (MI), which in general results from complications of atherosclerosis, is characterized by high inflammation and cardiomyocytes (CMs) apoptosis and by major loss of CMs. Regeneration of these lost CMs represents a major challenge for MI therapy. The increase of mitochondrial reactive oxygen species (ROS) is involved in cell cycle arrest which can be restarted by hypoxia or in the presence of ROS scavengers. Among ROS scavengers, mitochondrial thioredoxin 2 (Trx-2), an important antioxidant protein, could play a role in the CMs renewal. Method In this study, we investigated the effect of Trx-2 on mouse heart after an experimental MI. Results Trx-2 improves mouse cardiac function, reduces cardiac infarction size and increases the expression of cardiac anti-inflammatory markers. In addition, it reduces apoptosis and oxidative stress in heart tissue of mice after MI but it does not increase CM proliferation in cell culture or in heart tissue. Conclusion Mitochondrial Trx-2 effectively protects against heart infarction, likely via the reduction of oxidative stress, inflammation and apoptosis but not through CM renewal. Significance statement The current study unveils the complexities of MI and highlights mitochondrial Trx-2 role. Post-MI, marked by inflammation, CM apoptosis and significant CM loss. Trx-2 emerges as a vital protector. Its intervention improves mouse cardiac function, reduces infarction size and fosters an anti-inflammatory environment. By uncovering these mechanisms, the study suggests potential therapeutic strategies for oxidative stress, inflammation and apoptosis in MI, positioning Trx-2 as a promising candidate for future cardiac interventions.

Antioxidant and antiproliferative effect of a glycosaminoglycan extract from Rapana venosa marine snail.

Marine glycosaminoglycans (GAG) isolated from different invertebrates, such as molluscs, starfish or jellyfish, have been described as unique molecules with important pharmacological applications. Scarce information is available on GAG extract from Rapana venosa marine snail. The aim of this study was to isolate a GAG extract from R. venosa marine snail and to investigate its physicochemical, antioxidant and antiproliferative properties for further biomedical use. The morphology, chemical and elemental composition of the extract were established as well as the sulfate content and N- to O-sulfation ratio. Fourier transform infrared (FTIR) spectra indicated that GAG extract presented similar structural characteristics to bovine heparan sulfate and chondroitin sulfate. The pattern of extract migration in agarose gel electrophoresis and specific digestion with chondroitinase ABC and heparinase III indicated the presence of a mixture of chondroitin sulfate-type GAG, as main component, and heparan sulfate-type GAG. Free radical scavenging and ferric ion reducing assays showed that GAG extract had high antioxidant activity, which slightly decreased after enzymatic treatment. In vitro MTT and Live/Dead assays showed that GAG extract had the ability to inhibit cell proliferation in human Hep-2 cell cultures, at cytocompatible concentrations in normal NCTC clone L929 fibroblasts. This capacity decreased after enzymatic digestion, in accordance to the antioxidant activity of the products. Tumoral cell migration was also inhibited by GAG extract and its digestion products. Overall, GAG extract from R. venosa marine snail exhibited antioxidant and antiproliferative activities, suggesting its potential use as novel bioactive compound for biomedical applications.

Characterization and promotion of endothelialization of Bombyx mori silk fibroin functionalized with REDV peptide

In the development of small-diameter vascular grafts, it is crucial to achieve early-stage endothelialization to prevent thrombus formation and intimal hyperplasia. Silk fibroin (SF) from Bombyx mori is commonly used for such grafts. However, there is a need to expedite endothelialization post-implantation. In this study, we functionalized SF with Arg-Glu-Asp-Val (REDV) (SF + REDV) using cyanuric chloride to enhance endothelialization. The immobilization of REDV onto SF was confirmed and the amount of immobilized REDV could be calculated by 1H NMR. Furthermore, the conformational changes in Tyr, Ser, and Ala residues in [3-13C]Tyr- and [3-13C]Ser-SF due to REDV immobilization were monitored using 13C solid-state NMR. The REDV immobilized onto the SF film was found to be exposed on the film's surface, as confirmed by biotin-avidin system. Cell culture experiments, including adhesiveness, proliferation, and extensibility, were conducted using normal human umbilical vein endothelial cells (HUVEC) and normal human aortic smooth muscle cells (HAoSMC) on both SF and SF + REDV films to evaluate the impact of REDV on endothelialization. The results indicated a trend towards promoting HUVEC proliferation while inhibiting HAoSMC proliferation. Therefore, these findings suggest that SF + REDV may be more suitable than SF alone for coating small-diameter SF knitted tubes made of SF threads.

Improving the Efficacy of Common Cancer Treatments via Targeted Therapeutics towards the Tumour and Its Microenvironment.

Cancer is defined as the uncontrolled proliferation of heterogeneous cell cultures in the body that develop abnormalities and mutations, leading to their resistance to many forms of treatment. Left untreated, these abnormal cell growths can lead to detrimental and even fatal complications for patients. Radiation therapy is involved in around 50% of cancer treatment workflows; however, it presents significant recurrence rates and normal tissue toxicity, given the inevitable deposition of the dose to the surrounding healthy tissue. Chemotherapy is another treatment modality with excessive normal tissue toxicity that significantly affects patients' quality of life. To improve the therapeutic efficacy of radiotherapy and chemotherapy, multiple conjunctive modalities have been proposed, which include the targeting of components of the tumour microenvironment inhibiting tumour spread and anti-therapeutic pathways, increasing the oxygen content within the tumour to revert the hypoxic nature of the malignancy, improving the local dose deposition with metal nanoparticles, and the restriction of the cell cycle within radiosensitive phases. The tumour microenvironment is largely responsible for inhibiting nanoparticle capture within the tumour itself and improving resistance to various forms of cancer therapy. In this review, we discuss the current literature surrounding the administration of molecular and nanoparticle therapeutics, their pharmacokinetics, and contrasting mechanisms of action. The review aims to demonstrate the advancements in the field of conjugated nanomaterials and radiotherapeutics targeting, inhibiting, or bypassing the tumour microenvironment to promote further research that can improve treatment outcomes and toxicity rates.

Lanthanide-modified graphene oxide and nanodiamond materials and their cytotoxicity

Integrating lanthanide elements with carbon-based nanomaterials results in the creation of hybrid materials and nanocomposites with unique properties. We report a functionalization approach free of organic solvents for modifying graphene oxide (GO) and nanodiamond (ND) with various lanthanides (La(III), Ce(III), Eu(III), Gd(III), Tb(III) and Ho(III)). The aqueous-phase impregnation with their acetates was followed first by drying at 80 °C and then by heating at 300 °C for 2 h in air conditions. Scanning electron microscopy revealed that after the initial drying, the samples exhibited the presence of larger lanthanide-containing particles, whereas after the heating at 300 °C they became indistinguishable, and thus the lanthanide species became more homogeneously distributed throughout GO and ND. The toxicity, which is commonly associated with the presence of lanthanides, was evaluated in terms of the ability to inhibit the proliferation of COS-7 monkey kidney cell cultures. Most composites demonstrated a dose-dependent toxicity, with significantly lower cytotoxic effects compared to both the thermally treated GO and ND, as well as the pristine GO. The composites functionalized with gadolinium were among the least toxic at the maximum tested concentration (100 µg/ml). The ND-based composites were not cytotoxic at 50 µg/ml, the lowest tested concentration.

Thioredoxin-1 and its mimetic peptide improve systolic cardiac function and remodeling after myocardial infarction.

Myocardial infarction (MI) is characterized by a significant loss of cardiomyocytes (CMs), and it is suggested that reactive oxygen species (ROS) are involved in cell cycle arrest, leading to impaired CM renewal. Thioredoxin-1 (Trx-1) scavenges ROS and may play a role in restoring CM renewal. However, the truncated form of Trx-1, Trx-80, can compromise its efficacy by exerting antagonistic effects. Therefore, a Trx-1 mimetic peptide called CB3 was tested as an alternative way to restore CMs. This study aimed to investigate the effects of Trx-1, Trx-80, and CB3 on mice with experimental MI and study the underlying mechanism of CB3 on CMs. Mouse cardiac parameters were quantified by echocardiography, and infarction size and fibrosis determined using Trichrome and Picro-Sirius Red staining. The study found that Trx-1 and CB3 improved mouse cardiac function, reduced the size of cardiac infarct and fibrosis, and decreased the expression of cardiac inflammatory markers. Furthermore, CB3 polarized macrophages into M2 phenotype, reduced apoptosis and oxidative stress after MI, and increased CM proliferation in cell culture and invivo. CB3 effectively protected against myocardial infarction and could represent a new class of compounds for treating MI.

Ethanolic extract of &lt;em&gt;Aloe arborescens&lt;/em&gt; stimulates neonatal rat calvarial cells proliferation, migration and osteogenic differentiation

The medicinal plant Aloe arborescens Miller has chemical compounds that could stimulate the activity of bone-forming cells, but no studies have been found in this regard. We evaluated the effects of different dilutions of aqueous (1/10; 1/100; 1/1000) or ethanolic (1/1000; 1/2000; 1/5000) extracts of parenchyma from A. arborescens on the viability, proliferation, migration and osteogenic differentiation of primary cell cultures from neonatal rat calvaria. In none of the conditions studied did cell viability decrease (p ≤ 0.01). Furthermore, the 1/5000 ethanolic extract dilution showed a positive effect on cell viability at 48 h and 72 h and the latter was correlated with a 27 % (p ≤ 0.01) increase in cell proliferation. Ethanolic extract significantly stimulated cell migration and cultured mineralization with respect to control, showing the maximal effect at a dilution 1/5000. Together, the results show that the A. arborescens extracts do not have toxic effects. In addition, ethanolic extract stimulates proliferation, migration and osteogenic differentiation of rat calvarial cells, suggesting a potential bone anabolic action.