Modified parylene-N films as chemical microenvironments for differentiation and spheroid formation of osteoblast cells

Kim, Tae Hun; Lee, Jong‐Sook; Jo, Hanhee; Park, Yusun; Yun, Mijin; Song, Zhiquan; Pyun, Jae Chul; Misu, Lee

doi:10.1038/s41598-020-71322-1

Cited by 3 publications

(14 citation statements)

References 26 publications

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“…RNA was quantified using an ND-2000 spectrophotometer (Thermo Inc.). Total RNAs were extracted from HepG2 cells after histidine treatment under hypoxic conditions, as previously reported [ 29 ]. RNA sequencing and data analysis were assessed by EBIOGEN Inc (Seoul, Korea).…”

Section: Methodsmentioning

confidence: 99%

Impact of Exogenous Treatment with Histidine on Hepatocellular Carcinoma Cells

Park

Han

Kim

et al. 2022

Cancers

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Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. Sorafenib, a multi-kinase inhibitor, is the first-line therapy for advanced HCC. However, long-term exposure to sorafenib often results in reduced sensitivity and the development of resistance. Although various amino acids have been shown to contribute to cancer initiation and progression, little is known about the effects of histidine, a dietary essential amino acid that is partially taken up via histidine/large neutral amino acid transporter (LAT1), on cancer cells. In this study, we evaluated the effects of histidine on HCC cells and sensitivity to sorafenib. Remarkably, we found that exogenous histidine treatment induced a reduction in the expression of tumor markers related to glycolysis (GLUT1 and HK2), inflammation (STAT3), angiogenesis (VEGFB and VEGFC), and stem cells (CD133). In addition, LAT1 expression was downregulated in HCC tumor regions with high expression of GLUT1, CD133, and pSTAT3, which are known to induce sorafenib resistance. Finally, we demonstrated that combined treatment with sorafenib and histidine could be a novel therapeutic strategy to enhance the sensitivity to sorafenib, thereby improving long-term survival in HCC.

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Section: Methodsmentioning

confidence: 99%

Impact of Exogenous Treatment with Histidine on Hepatocellular Carcinoma Cells

Park

Han

Kim

et al. 2022

Cancers

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“…UV-treated parylene-N and UV-treated parylene-C films were also prepared to include oxygen species on the surface, such as hydroxyl, formyl, and carboxyl groups. [6][7][8]28,45,46 For the seven different surfaces (6 parylene films and polystyrene as control), the roughness of the films was analyzed using AFM (Figure 1B), and represented as average roughness ± standard deviation (%). The estimated roughness for parylene films were found tobe parylene-N (5.93 nm ±4.2%), UV-treated parylene-N (5.6 nm ±8.1%), parylene-C (2.3 nm ±7.0%), UV-treated parylene-C (4.0 nm ±4.0%), parylene-AM (1.5 nm ±16.0%), and parylene-H (3.2 nm ±14.2%).…”

Section: Resultsmentioning

confidence: 99%

“…For measuring the surface roughness, three random areas (5 × 5 μm) on parylene films were analyzed using noncontact mode AFM, and the surface roughness was calculated from the root-mean-square roughness ( R q ) of the surface. ,− To measure the contact angles, water drops (10 μL each) were dispensed on five different random areas of the parylene films and measured using an automated system (SmartDrop; Femtofab Co., Seoul, South Korea). Contact angle analysis of the water drop was performed immediately after the drop was dispensed. , To measure the surface energy, a diiodomethane (DIM) drop (1 μL) was initially dispensed in five different random areas of the parylene films. Contact angle analysis was performed immediately after the DIM drop.…”

Section: Methodsmentioning

confidence: 99%

“…Functionalized parylene films have been used to immobilize proteins with a wide range of isoelectric point (pI) values (from acidic to basic); they are typically used for the covalent immobilization of peptides, enzymes, and antibodies through chemical functional groups, and their immobilization efficiency is significantly enhanced compared to physical adsorption, especially for small proteins such as peptides . Functionalized parylenes have also been effectively used in cell culture and spheroid production . An osteoblast-like cell, MG-63, was cultured on parylene-N film, and considerable changes were observed in cell adhesion and cell viability before and after UV treatment .…”

Section: Introductionmentioning

confidence: 99%

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Functionalized Parylene Films for Enhancement of Antibody Production by Hybridoma Cells

Kim,

Song,

Jung

et al. 2023

ACS Appl. Bio Mater.

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In this study, the influence of microenvironments on antibody production of hybridoma cells was analyzed using six types of functionalized parylene films, parylene-N and parylene-C (before and after UV radiation), parylene-AM, and parylene-H, and using polystyrene as a negative control. Hybridoma cells were cultured on modified parylene films that produced a monoclonal antibody against the well-known fungal toxin ochratoxin-A. Surface properties were analyzed for each parylene film, such as roughness, chemical functional groups, and hydrophilicity. The proliferation rate of the hybridoma cells was observed for each parylene film by counting the number of adherent cells, and the total amount of produced antibodies from different parylene films was estimated using indirect ELISA. In comparison with the polystyrene, the antibody-production by parylene-H and parylene-AM was estimated to be observed to be as high as 210–244% after the culture of 24 h. These results indicate that the chemical functional groups of the culture plate could influence antibody production. To analyze the influence of the microenvironments of the modified parylene films, we performed cell cycle analysis to estimate the ratio of the G0/G1, S, and G2/M phases of the hybridoma cells on each parylene film. From the normalized proportion of phases of the cell cycle, the difference in antibody production from different surfaces was considered to result from the difference in the proliferation rate of hybridoma cells, which occurred from the different physical and chemical properties of the parylene films. Finally, protein expression was analyzed using an mRNA array to determine the effect of parylene films on protein expression in hybridoma cells. The expression of three antibody production-related genes (CD40, Sox4, and RelB) was analyzed in hybridoma cells cultured on modified parylene films.

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“…In this experiment, IL-1 β markedly stimulated rat osteoblast proliferation, which was inhibited by GDC0623 treatment. Elevated osteoblast proliferation levels enhance bone formation during normal bone remodeling [ 26 ]. However, IL-1 β treatment inhibited osteogenic differentiation.…”

Section: Discussionmentioning

confidence: 99%

MEK1/2 Inhibitor (GDC0623) Promotes Osteogenic Differentiation of Primary Osteoblasts Inhibited by IL-1β through the MEK-Erk1/2 and Jak/Stat3 Pathways

Zhang

et al. 2021

International Journal of Endocrinology

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Objective. We evaluated the effects and mechanisms of GDC0623 on osteogenic differentiation of osteoblasts induced by IL-1β. Methodology. Osteoblasts were treated with 20 ng/ml IL-1β and 0.1 µM GDC0623. Cell proliferation levels were evaluated by the cell counting kit 8 (CCK8), EdU assay, and western blotting [proliferating cell nuclear antigen (PCNA) and Cyclin D1]. Osteoblasts were cultured in an osteogenic induction medium for 1–3 weeks after which their differentiations were assessed by alkaline phosphatase (ALP) staining, Alizarin Red staining, calcium concentration, immunocytochemistry staining, real-time quantitative PCR (RT-qPCR), and immunofluorescence staining. The osteogenesis-associated mechanisms were further evaluated by western blotting using appropriate antibodies. Results. Relative to the control group, IL-1β induced the rapid proliferation of osteoblasts and suppressed their osteogenic differentiations by upregulating the activities of MEK-Erk1/2 as well as Jak-Stat3 pathways and by elevating MMP13 and MMP9 levels. However, blocking of the MEK-Erk1/2 signaling pathway by GDC0623 treatment reversed these effects. Conclusion. Inhibition of Jak-Stat3 pathway by C188-9 downregulated the expression levels of MMP9 and MMP13, activated MEK-Erk1/2 pathway, and inhibited osteogenic differentiation.

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Modified parylene-N films as chemical microenvironments for differentiation and spheroid formation of osteoblast cells

Cited by 3 publications

References 26 publications

Impact of Exogenous Treatment with Histidine on Hepatocellular Carcinoma Cells

Impact of Exogenous Treatment with Histidine on Hepatocellular Carcinoma Cells

Functionalized Parylene Films for Enhancement of Antibody Production by Hybridoma Cells

MEK1/2 Inhibitor (GDC0623) Promotes Osteogenic Differentiation of Primary Osteoblasts Inhibited by IL-1β through the MEK-Erk1/2 and Jak/Stat3 Pathways

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