scholarly journals Electrospun Fibres with Hyaluronic Acid-Chitosan Nanoparticles Produced by a Portable Device

Nanomaterials ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 2016
Author(s):  
Carla V. Fuenteslópez ◽  
Hua Ye
Keyword(s):  
Cell Proliferation ◽  
Hyaluronic Acid ◽  
Cell Viability ◽  
Cell Attachment ◽  
Chitosan Nanoparticles ◽  
Cell Interaction ◽  
Portable Device ◽  
Growth And Yield ◽  
Directional Growth ◽  
Versatile Technique

Electrospinning is a versatile technique to produce nano/microscale fibrous scaffolds for tissue engineering and drug delivery applications. This research aims to demonstrate that hyaluronic acid-chitosan (HA-CS) nanoparticles can be electrospun together with polycaprolactone (PCL) and gelatine (Ge) fibres using a portable device to create scaffolds for tissue repair. A range of polymer solutions of PCL-gelatine at different weight/volume concentrations and ratios were electrospun and characterised. Fibre–cell interaction (F11 cells) was evaluated based on cell viability and proliferation and, from here, a few polymer blends were electrospun into random or aligned fibre arrangements. HA-CS nanoparticles were synthesised, characterised, and used to functionalise electrospun fibres (8% w/v at 70 PCL:30 Ge), which were chosen based on cell viability. Different concentrations of HA-CS nanoparticles were tested to determine cytotoxicity. A single dosage (1 × 10−2 mg/mL) was associated with higher cell proliferation compared with the cell-only control. This nanoparticle concentration was embedded into the electrospun fibres as either surface modification or blend. Fibres with blended NPs delivered a higher cell viability than unmodified fibres, while NP-coated fibres resulted in a higher cell proliferation (72 h) than the NP-blended ones. These biocompatible scaffolds allow cell attachment, maintain fibre arrangement, promote directional growth and yield higher cell viability.

scholarly journals Development of the osteoblast phenotype of serial cell subcultures from human bone marrow

2005 ◽  
Vol 16 (3) ◽  
pp. 225-230 ◽  
Author(s):  
Adalberto Luiz Rosa ◽  
Márcio Mateus Beloti
Keyword(s):  
Bone Marrow ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Protein Content ◽  
Total Protein ◽  
Bone Marrow Cells ◽  
Cell Attachment ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Total Protein Content

Bone marrow cells have been used for testing biocompatibility of bone substitute materials that would be applied in maxillofacial and orthopedic surgeries. However, it remains unclear whether cells in serial subcultures retain the ability to differentiate into osteoblasts. The purpose of this study was to compare the development of osteoblast phenotype of serially passaged cells from human bone marrow. Cells from first to third passage were cultured (2x10(4) cells/well) in supplemented culture medium. Cells were incubated at 37ºC in a humidified atmosphere of 5% CO2 and 95% air. Cell attachment was assessed at 4 and 24 h. At 7, 14 and 21 days, cell proliferation, cell viability, total protein content and alkaline phosphatase (ALP) activity were evaluated. Bone-like formation was evaluated at 14 and 21 days. Data were compared by two-way ANOVA and Duncan's multiple range test. Cell attachment, cell viability and total protein content were not affected by serial subcultures. However, serial subcultures did interfered negatively with osteoblast differentiation as shown by osteoblast parameters observed in second and third subcultures, such as continuous cell proliferation, lower ALP activity and bone-like formation in comparison to first subculture. Therefore, it is important to evaluate cell ability to growth and differentiate before selecting the cell population for studies that investigate the biocompatibility of materials to replace bone tissue.

scholarly journals Cytotoxic Effect of Chitosan Nanoparticles on Normal Human Dental Pulp Cells

2019 ◽  
Vol 3 (1) ◽  
Author(s):  
Rami Alhomrany ◽  
Chang Zhang ◽  
Laisheng Chou
Keyword(s):  
Cell Viability ◽  
Dental Pulp ◽  
Cell Attachment ◽  
Chitosan Nanoparticles ◽  
Dental Pulp Cells ◽  
Human Dental Pulp Cells ◽  
Attachment Efficiency ◽  
Human Dental Pulp ◽  
Pulp Cells ◽  
Normal Human

 Introduction: Recent in vitro studies have shown that chitosan nanoparticles could enhance the antimicrobial activity of several dental materials. However, the biocompatibility of these nanoparticles with normal human cells is still controversial. The aim of this study was to evaluate the potential toxicity of various sizes and concentrations of chitosan nanoparticles cultured with normal human dental pulp cells. Methods: Normal human dental pulp cells were derived from human dental pulp tissues and cultured with (50-67) nm and (318-350) nm chitosan nanoparticles in concentrations: 0.2 mg/mL, 0.5 mg/mL, 1 mg/mL, and 2 mg/mL as study groups, and 0 mg/mL as a control. The cell attachment efficiency for each group was assessed at 16 hours. The proliferation rate and cell viability were evaluated at days 7 and 14. Both, attachment efficiency and proliferation rate were assessed by measuring the optical density of crystal violet stained cells. The cell viability was determined by the activity of the mitochondrial dehydrogenase enzyme. Statistical analysis was performed using One-Way ANOVA and post hoc Tukey test. Results: All concentrations of the (50-67) nm group significantly reduced cell attachment efficiency in comparison with the control (p<0.01) and with the (318-350) nm group (p<0.01). All concentrations of both groups, (50-67) nm and (318-350) nm, significantly reduced cell proliferation and cell viability compared to the control in dose-dependent and size-associated manners. (p<0.01).    Conclusion: Chitosan nanoparticles exhibit a cytotoxic effect on normal human dental pulp cells

Comparison of Three Microstructure Fabrication Methods for Bone Cell Growth Studies

2008 ◽  
Author(s):  
Marzellus Große Holthaus ◽  
Kurosch Rezwan
Keyword(s):  
Cell Proliferation ◽  
Cell Growth ◽  
Microcontact Printing ◽  
Cell Interaction ◽  
Bone Cell ◽  
Directional Growth ◽  
Growth Studies ◽  
Optical Profilometer ◽  
Complete Cell ◽  
Aerosol Jet Printing

Different micropatterning techniques were applied to elucidate the potential for cell proliferation studies on calcium phosphate surfaces. Sintered hydroxyapatite (HA) platelets were microstructured by three different techniques: Aerosol jet printing (M3D®), laser ablation and microcontact printing via polydimethylsiloxane (PDMS) stamps. The microstructures were designed as channels between 1000 and 3000 micron in length, 10 to 220 micron in width and 5 to 110 micron in height. An optical profilometer, a Scanning Electron Microscope (SEM) and X-ray diffraction were used to characterize the microstructures. Cell proliferation tests were carried out by incubating the microstructured ceramic samples in complete cell media for a maximum of seven days. Osteoblast-like cells (MG-63) were used for testing. Each sample was immersed in media in which the cells were already seeded. Imaging was performed by SEM and Fluorescence Microscopy. The cells proliferated on all three differently fabricated microstructures. Cell growth was observed in the microchannels as well as on the microchannel walls or spacers. In particular it turned out, that the microtopology can provoke the cells to elongate aligned to the direction of the microchannels. Non-directional growth was observed on non-structured areas. All three differently fabricated hydroxyapatite microstructuring methods seem to be attractive and promising techniques for use in bone cell growth studies. The applied fabrication techniques show many advantages for fundamental research in the field of cell interaction with ceramic microstructures and may exhibit possible methods of structuring implant surfaces.

scholarly journals Photocuring Hyaluronic Acid/Silk Fibroin Hydrogel Containing Curcumin Loaded CHITOSAN Nanoparticles for the Treatment of MG-63 Cells and ME3T3-E1 Cells

Polymers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 2302
Author(s):  
Qingwen Yu ◽  
Zhiyuan Meng ◽  
Yichao Liu ◽  
Zehao Li ◽  
Xing Sun ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Bone Regeneration ◽  
Water Uptake ◽  
Silk Fibroin ◽  
Bone Repair ◽  
Ph Value ◽  
Chitosan Nanoparticles ◽  
Vitro Assay ◽  
Long Term Treatment

After an osteosarcoma excision, recurrence and bone defects are significant challenges for clinicians. In this study, the curcumin (Cur) loaded chitosan (CS) nanoparticles (CCNP) encapsulated silk fibroin (SF)/hyaluronic acid esterified by methacrylate (HAMA) (CCNPs-SF/HAMA) hydrogel for the osteosarcoma therapy and bone regeneration was developed by photocuring and ethanol treatment. The micro or nanofibers networks were observed in the CCNPs-SF/HAMA hydrogel. The FTIR results demonstrated that alcohol vapor treatment caused an increase in β-sheets of SF, resulting in the high compression stress and Young’s modulus of CCNPs-SF/HAMA hydrogel. According to the water uptake analysis, SF caused a slight decrease in water uptake of CCNPs-SF/HAMA hydrogel while CCNPs could enhance the water uptake of it. The swelling kinetic results showed that both the CCNPs and the SF increased the swelling ratio of CCNPs-SF/HAMA hydrogel. The accumulative release profile of CCNPs-SF/HAMA hydrogel showed that the release of Cur from CCNPs-SF/HAMA hydrogel was accelerated when pH value was decreased from 7.4 to 5.5. Besides, compared with CCNPs, the CCNPs-SF/HAMA hydrogel had a more sustainable drug release, which was beneficial for the long-term treatment of osteosarcoma. In vitro assay results indicated that CCNPs-SF/HAMA hydrogel with equivalent Cur concentration of 150 μg/mL possessed both the effect of anti-cancer and promoting the proliferation of osteoblasts. These results suggest that CCNPs-SF/HAMA hydrogel with superior physical properties and the bifunctional osteosarcoma therapy and bone repair may be an excellent candidate for local cancer therapy and bone regeneration.

scholarly journals Wnt5A modulates integrin expression in a receptor-dependent manner in ovarian cancer cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Vajihe Azimian-Zavareh ◽  
Zeinab Dehghani-Ghobadi ◽  
Marzieh Ebrahimi ◽  
Kian Mirzazadeh ◽  
Irina Nazarenko ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Cell Attachment ◽  
Ovarian Cancer Cells ◽  
Integrin Expression ◽  
Dependent Manner ◽  
Cancer Cell Proliferation ◽  
Expression Levels ◽  
Multicellular Aggregates ◽  
Focal Adhesion Kinase Activity

AbstractWnt5A signals through various receptors that confer versatile biological functions. Here, we used Wnt5A overexpressing human ovarian SKOV-3 and OVCAR-3 stable clones for assessing integrin expression, cell proliferation, migration, invasion, and the ability of multicellular aggregates (MCAs) formation. We found here, that Wnt5A regulates differently the expression of its receptors in the stable Wnt5A overexpressing clones. The expression levels of Frizzled (FZD)-2 and -5, were increased in different clones. However ROR-1, -2 expression levels were differently regulated in clones. Wnt5A overexpressing clones showed increased cell proliferation, migration, and clonogenicity. Moreover, Wnt5A overexpressing SKOV-3 clone showed increased MCAs formation ability. Cell invasion had been increased in OVCAR-3-derived clones, while this was decreased in SKOV-3-derived clone. Importantly, αv integrin expression levels were increased in all assessed clones, accompanied by increased cell attachment to fibronectin and focal adhesion kinase activity. Moreover, the treatment of clones with Box5 as a Wnt5A/FZD5 antagonist abrogates ITGAV increase, cell proliferation, migration, and their attachment to fibronectin. Accordingly, we observed significantly higher expression levels of ITGAV and ITGB3 in human high-grade serous ovarian cancer specimens and ITGAV correlated positively with Wnt5A in metastatic serous type ovarian cancer. In summary, we hypothesize here, that Wnt5A/FZD-5 signaling modulate αv integrin expression levels that could be associated with ovarian cancer cell proliferation, migration, and fibronectin attachment.

scholarly journals Optimization of Spheroid Colony Culture and Cryopreservation of Nucleus Pulposus Cells for the Development of Intervertebral Disc Regenerative Therapeutics

2021 ◽  
Vol 11 (8) ◽  
pp. 3309
Author(s):  
Kosuke Sako ◽  
Daisuke Sakai ◽  
Yoshihiko Nakamura ◽  
Erika Matsushita ◽  
Jordy Schol ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Viability ◽  
Nucleus Pulposus ◽  
Mass Culture ◽  
Formation Rate ◽  
Proliferation Rate ◽  
Cell Phenotype ◽  
Mrna Levels ◽  
Spheroid Formation ◽  
Positive Nucleus

After the discovery of functionally superior Tie2-positive nucleus pulposus (NP) progenitor cells, new methods were needed to enable mass culture and cryopreservation to maintain these cells in an undifferentiated state with high cell yield. We used six types of EZSPHERE® dishes, which support spheroid-forming colony culture, and examined NP cell spheroid-formation ability, number, proliferation, and mRNA expression of ACAN, COL1A2, COL2A1, and ANGPT1. Six different types of cryopreservation solutions were examined for potential use in clinical cryopreservation by comparing the effects of exposure time during cryopreservation on cell viability, Tie2-positivity, and cell proliferation rates. The spheroid formation rate was 45.1% and the cell proliferation rate was 7.75 times using EZSPHERE® dishes. The mRNA levels for COL2A1 and ANGPT1 were also high. In cryopreservation, CryoStor10 (CS10) produced ≥90% cell viability and a high proliferation rate after thawing. CS10 had a high Tie2-positive rate of 12.6% after culturing for 5 days after thawing. These results suggest that EZSPHERE enabled colony formation in cell culture without the use of hydrogel products and that CS10 is the best cryopreservation medium for retaining the NP progenitor cell phenotype and viability. Together, these data provide useful information of NP cell-based therapeutics to the clinic.

scholarly journals The Olive Leaves Extract Has Anti-Tumor Effects against Neuroblastoma through Inhibition of Cell Proliferation and Induction of Apoptosis

Nutrients ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 2178
Author(s):  
Fabio Morandi ◽  
Veronica Bensa ◽  
Enzo Calarco ◽  
Fabio Pastorino ◽  
Patrizia Perri ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Cell Cycle Progression ◽  
Treatment Strategies ◽  
Annexin V ◽  
Dependent Manner ◽  
Induction Of Apoptosis ◽  
Dose Dependent

Neuroblastoma (NB) is the most common extra-cranial solid tumor of pediatric age. The prognosis for high-risk NB patients remains poor, and new treatment strategies are desirable. The olive leaf extract (OLE) is constituted by phenolic compounds, whose health beneficial effects were reported. Here, the anti-tumor effects of OLE were investigated in vitro on a panel of NB cell lines in terms of (i) reduction of cell viability; (ii) inhibition of cell proliferation through cell cycle arrest; (iii) induction of apoptosis; and (iv) inhibition of cell migration. Furthermore, cytotoxicity experiments, by combining OLE with the chemotherapeutic topotecan, were also performed. OLE reduced the cell viability of NB cells in a time- and dose-dependent manner in 2D and 3D models. NB cells exposed to OLE underwent inhibition of cell proliferation, which was characterized by an arrest of the cell cycle progression in G0/G1 phase and by the accumulation of cells in the sub-G0 phase, which is peculiar of apoptotic death. This was confirmed by a dose-dependent increase of Annexin V+ cells (peculiar of apoptosis) and upregulation of caspases 3 and 7 protein levels. Moreover, OLE inhibited the migration of NB cells. Finally, the anti-tumor efficacy of the chemotherapeutic topotecan, in terms of cell viability reduction, was greatly enhanced by its combination with OLE. In conclusion, OLE has anti-tumor activity against NB by inhibiting cell proliferation and migration and by inducing apoptosis.

NEAT1/miR-140-3p/MAPK1 mediates the viability and survival of coronary endothelial cells and affects coronary atherosclerotic heart disease

2020 ◽  
Vol 52 (9) ◽  
pp. 967-974
Author(s):  
Hui Zhang ◽  
Ningning Ji ◽  
Xinyan Gong ◽  
Shimao Ni ◽  
Yu Wang
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cells ◽  
Heart Disease ◽  
Cell Viability ◽  
Mitogen Activated Protein Kinase ◽  
Expression Level ◽  
Luciferase Reporter ◽  
Coronary Endothelial Cells ◽  
Induced Apoptosis ◽  
Lncrna Neat1

Abstract Studies have shown that long non-coding RNAs (lncRNA) play critical roles in coronary atherosclerotic heart disease (CAD). However, the function of lncRNA nuclear enriched abundant transcript 1 (NEAT1) in CAD is unclear. In this study, we aimed to investigate the functions of lncRNA NEAT1 in CAD. RT-PCR and western blot analysis were carried out to examine the expressions of related RNAs. Colony formation assay, cell proliferation assay, apoptosis assay, and dual-luciferase reporter assay were conducted to investigate the abilities of colony migration, cell proliferation, apoptosis, and targeting. The results showed that NEAT1 was up-regulated in CAD blood samples and in human coronary endothelial cells (HCAECs). Transfection of pcNEAT1 significantly inhibited the survival rate of HCAECs and induced apoptosis of HCAECs. MiR-140-3p was down-regulated in HCAECs. NEAT1 directly targeted miR-140-3p, and the expression of miR-140-3p was inversely correlated with the expression of NEAT1 in CAD patients. In addition, co-transfection of NEAT1 with miR-140-3p mimic reversed the effect of pcNEAT1 on cell viability and apoptosis. mitogen-activated protein kinase 1 (MAPK1) was proved to be a target gene of miR-140-3p, and the miR-140-3p mimic was shown to reduce the expression of MAPK1 in HCAECs. pcNEAT1 significantly increased the expression level of MAPK1, while shNEAT1 significantly reduced the expression level of MAPK1. Our results revealed that lncRNA NEAT1 increased cell viability and inhibited CAD cell apoptosis possibly by activating the miR-140-3p/MAPK1 pathway, and lncRNA NEAT1 might serve as a potential therapeutic target for CAD.

scholarly journals Amygdalin promotes the activity of T cells to suppress the progression of HBV-related hepatocellular carcinoma via the JAK2/STAT3 signaling pathway

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Ruoyu Wang ◽  
Dong Zhang ◽  
Kewei Sun ◽  
Jianping Peng ◽  
Wenfang Zhu ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Proliferation ◽  
T Cells ◽  
T Cell ◽  
Cell Viability ◽  
Caspase 3 ◽  
Invasion And Migration ◽  
Ifn Γ ◽  
And Migration ◽  
Cellular Immune

Abstract Background Hepatitis B virus (HBV) infection is a high-risk factor of hepatocellular carcinoma (HCC). Cellular immune responses are essential for HCC development, and the CD4+ and CD8+ T subtypes are identified as the primary anti-tumor immune cells. In the study, we investigated the effect and mechanism of amygdalin in the cellular immune response in HBV-related HCC and HCC progression. Methods The cell proliferation was examined by MTT analysis. Cells metastasis ability was detected by Invasion and migration assays. Quantification of apoptotic cells was performed with Flow cytometer assay. The protein levels of p-STAT3, STAT3, p-JAK2, JAK2, caspase-3, cleaved caspase-3 were detected by performing immunoblotting assays. Results We demonstrate that amygdalin treatment could rescue the HBV-T cell viability and IFN-γ and TNF-αproduction. In HBV-T cells, the MFI levels of CD8+ are lower than that in NC-T cells. Moreover, the phosphorylation levels of STAT3 and JAK2 are higher in HBV-T cells, compared to those in NC-T cells, and then reduced by amygdalin treatment. Co-culture with HBV-T cells could reduce IFN-γ and TNF-α, production while increase IL-6 and IL-10 production in HepG2.2.15 cells; these alterations could be partially reversed by amygdalin pretreatment. Finally, co-culture with HBV-T cells significantly promoted the cell viability, inhibited the apoptosis, and promoted the migration of HepG2.2.15 cells, and these alterations could be partially reversed by amygdalin treatment. Conclusion Our findings provide a rationale for further studies on the functions and mechanism of amygdalin inhibiting HBV-related HCC cell proliferation, invasion, and migration via T cell-mediated tumor immunity.

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