Prednisolone succinate–glucosamine conjugate: Synthesis, characterization and in vitro cellular uptake by kidney cell lines

e13528 Background: A number of studies have reported the superior antitumor effect of nanoparticles loading chemotherapeutics than the free agents, yet the underlying mechanism has not attract enough attention. The extracellular pH of cancer cells is lower than that of the intracellular pH. Due to this pH gradient, weakly basic drug will protonated extracellularly and display decreased intracellular concentration. In this study, we aimed to reveal a new mechanism of PEG-PCL nanoparticles, namely the reversion of physiological drug resistance. Methods: Tetradrine (Tet), an alkaloid isolated from traditional Chinese medicine, was incorporated into the diblock copolymer methoxy poly(ethylene glycol)-polycaprolactone (mPEG-PCL). In vitro cytotoxicity of free Tet and Tet-loaded nanoparticles at pH7.4 and pH6.8 was compared on four different cancer cell lines. Fluorescent particle cellular uptake study was also used. To evaluate the antitumor effect of the nanoparticles in a more complex model rather than monolayer cell culture, we used Histoculture Drug Resistance Assay (HDRA). The in vivo antitumor effect of the nanoparticles was also studied in ICR mice bearing H22 tumor with different in vivo pH values. Results: In vitro cytotoxicity study in four tumor cell lines showed that the cytotoxicity of free Tet decreased significantly (P<0.05) when the extracellular pH decreased from 7.4 to 6.8, while the cytotoxicity of Tet-loaded nanoparticles increased or didn’t change significantly. The possible mechanism may mainly be the endocytosis of nanoparticles, which was proven by fluorescent particle cellular uptake study. HDRA indicated better tissue penetration of nanoparticles over free Tet. As to in vivo study, the mice with in vivo tumor pH 6.8 and treated with Tet-loaded nanoparticles exhibited best tumor inhibit rate and mildest side effect, suggesting that the use of nanoparticles was more preferable than the manipulation of tumor pH by the use of basic water. Conclusions: Our study clearly demonstrated that the mPEG-PCL nanoparticles could overcome the drug resistance caused by low extracellular pH and enhance drug penetration in the tumor tissue, thus increasing the antitumor efficacy of weakly basic agents. No significant financial relationships to disclose.

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In vitro cytotoxicity of two novel oral formulations of Amphotericin B (iCo-009 and iCo-010) against Candida albicans, human monocytic and kidney cell lines

Lipids in Health and Disease ◽

10.1186/1476-511x-10-144 ◽

2011 ◽

Vol 10 (1) ◽

pp. 144 ◽

Cited By ~ 10

Author(s):

Carlos G Leon ◽

Jinkyung Lee ◽

Karen Bartlett ◽

Pavel Gershkovich ◽

Ellen K Wasan ◽

...

Keyword(s):

Candida Albicans ◽

Amphotericin B ◽

Cell Lines ◽

Kidney Cell ◽

In Vitro Cytotoxicity ◽

Oral Formulations

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In vitro proof of concept studies of radiotoxicity from Auger electron-emitter thallium-201

EJNMMI Research ◽

10.1186/s13550-021-00802-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Katarzyna M. Osytek ◽

Philip J. Blower ◽

Ines M. Costa ◽

Gareth E. Smith ◽

Vincenzo Abbate ◽

...

Keyword(s):

Dna Damage ◽

Cell Lines ◽

Cellular Uptake ◽

Auger Electron ◽

Nuclear Dna ◽

Thallium 201 ◽

Cell Uptake ◽

High Potassium ◽

Du145 Cells

Abstract Background Auger electron-emitting radionuclides have potential in targeted treatment of small tumors. Thallium-201 (201Tl), a gamma-emitting radionuclide used in myocardial perfusion scintigraphy, decays by electron capture, releasing around 37 Auger and Coster–Kronig electrons per decay. However, its therapeutic and toxic effects in cancer cells remain largely unexplored. Here, we assess 201Tl in vitro kinetics, radiotoxicity and potential for targeted molecular radionuclide therapy, and aim to test the hypothesis that 201Tl is radiotoxic only when internalized. Methods Breast cancer MDA-MB-231 and prostate cancer DU145 cells were incubated with 200–8000 kBq/mL [201Tl]TlCl. Potassium concentration varied between 0 and 25 mM to modulate cellular uptake of 201Tl. Cell uptake and efflux rates of 201Tl were measured by gamma counting. Clonogenic assays were used to assess cell survival after 90 min incubation with 201Tl. Nuclear DNA damage was measured with γH2AX fluorescence imaging. Controls included untreated cells and cells treated with decayed [201Tl]TlCl. Results 201Tl uptake in both cell lines reached equilibrium within 90 min and washed out exponentially (t1/2 15 min) after the radioactive medium was exchanged for fresh medium. Cellular uptake of 201Tl in DU145 cells ranged between 1.6 (25 mM potassium) and 25.9% (0 mM potassium). Colony formation by both cell lines decreased significantly as 201Tl activity in cells increased, whereas 201Tl excluded from cells by use of high potassium buffer caused no significant toxicity. Non-radioactive TlCl at comparable concentrations caused no toxicity. An estimated average 201Tl intracellular activity of 0.29 Bq/cell (DU145 cells) and 0.18 Bq/cell (MDA-MB-231 cells) during 90 min exposure time caused 90% reduction in clonogenicity. 201Tl at these levels caused on average 3.5–4.6 times more DNA damage per nucleus than control treatments. Conclusions 201Tl reduces clonogenic survival and increases nuclear DNA damage only when internalized. These findings justify further development and evaluation of 201Tl therapeutic radiopharmaceuticals.

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Trophic factor secreting kidney cell lines: in vitro characterization and functional effects following transplantation in ischemic rats

Brain Research ◽

10.1016/s0006-8993(01)02327-7 ◽

2001 ◽

Vol 900 (2) ◽

pp. 268-276 ◽

Cited By ~ 38

Author(s):

Rowena E Johnston ◽

Ora Dillon-Carter ◽

William J Freed ◽

Cesario V Borlongan

Keyword(s):

Cell Lines ◽

Kidney Cell ◽

Trophic Factor ◽

In Vitro Characterization

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IN VITRO CYTOTOXICITY ACTIVITY OF HYDROALCOHOLIC EXTRACT OF POLYHERBAL CHURNAM (COFRI) IN NORMAL LIVER AND KIDNEY CELL LINES

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2019.v12i3.26971 ◽

2019 ◽

pp. 74-77

Author(s):

YASMINE Y ◽

CAROLINE JEBA R

Keyword(s):

Cell Lines ◽

Kidney Cell ◽

Colorimetric Assay ◽

Normal Liver ◽

In Vitro Cytotoxicity ◽

Human Consumption ◽

Normal Liver Cell ◽

Hydroalcoholic Extract ◽

Liver And Kidney

Objective: The objective of the study was to investigate the cytotoxic activity of hydroalcoholic extract of a polyherbal powder on Vero (normal kidney) and Chang (normal liver) cell lines. Methods: The cytotoxic effect was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide assay. It is commonly used colorimetric assay for assessing cell metabolic activity under specific conditions, indicates the number of viable cells. Results: According to the experimental results, the hydroalcoholic extract does not cause any toxicity on normal liver and kidney cell lines and LC50 values are ca lculated using LDP line software and found 776.73 μg/ml and 686.58 μg/ml. Conclusion: The hydroalcoholic extract of polyherbal churnam exhibited cell viability at higher concentrations. The results indicates investigated formulation does not causes any toxicity to the normal liver and kidney cells and found to be safe for human consumption. Further, toxicological and pharmacological evaluation is required to prove its safety and efficacy.

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Investigating the mechanisms of indocyanine green (ICG) cellular uptake in sarcoma

10.1101/2021.04.05.438013 ◽

2021 ◽

Author(s):

Corey D Chan ◽

Marcus J Brookes ◽

Riya Tanwani ◽

Chloe Hope ◽

Toni A Pringle ◽

...

Keyword(s):

Indocyanine Green ◽

Cell Line ◽

Cell Lines ◽

Cellular Uptake ◽

Incubation Time ◽

Near Infrared ◽

Cancer Cell Line ◽

Proliferation Rate ◽

Low Grade

Introduction: Indocyanine green (ICG) is a near infrared (NIR) dye which has been used clinically for over 50 years and has recently been utilised for fluorescence guided surgery in a number of cancer types, including sarcoma. ICG is taken up and retained by sarcoma tumours to a greater extent than normal tissue, demonstrating its potential to aid in visualisation of tumour margins. However, the mechanisms surrounding preferential ICG uptake in tumours are poorly understood. Methods: In vitro ICG cellular uptake studies were performed across a panel of four sarcoma cell lines and one breast cancer cell line, exhibiting varying proliferation rates and phenotypes. The effects of ICG concentration, incubation time, inhibition of clathrin mediated endocytosis and cell line proliferation rate on the cellular uptake of ICG were investigated, using fluorescence microscopy and flow cytometry. The spatial orientation of ICG was also assessed in a patient specimen. Results: The level of ICG cellular uptake was dependent on ICG concentration and incubation time. Cell line proliferation rate correlated significantly to ICG uptake within 30 minutes (Rs = 1, p<0.001), whilst retention of ICG after 24hrs did not (Rs = 0.3, p=0.624). From our data, the primary mechanism of ICG uptake in sarcoma cells is via clathrin mediated endocytosis. Following the resection of a grade 3 leiomyosarcoma, ICG signal was detectable macroscopically and on 3um sections, whilst being negative on the muscle control. Conclusions: The use of ICG for tumour detection in sarcoma surgery may demonstrate higher utility in high grade tumours compared to low grade tumours, due to the observation of higher ICG uptake in more proliferative cell lines. It is likely that the enhanced permeability and retention (EPR) effect plays a significant role in the retention of ICG within tumours. Future work on the detection of ICG at the cellular level within human tissue sections is required, with the aid of purpose built NIR microscopes.

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EVALUATION OF IN VITRO CYTOTOXIC AND ANTICANCER ACTIVITY OF HEPATOPROTECTIVE POLYHERBAL FORMULATION IN CELL LINE MODEL

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2019.v12i4.32118 ◽

2019 ◽

pp. 320-325

Author(s):

JAYACHANDRA KUNCHA ◽

THIRUGNANASAMBANTHAM P ◽

KUMARAN S ◽

NARAYANAN N ◽

SHARMILA DEVI V

Keyword(s):

Anticancer Activity ◽

Cell Lines ◽

Cancer Cell ◽

Anticancer Agents ◽

Kidney Cell ◽

Normal Liver ◽

Cancer Cell Lines ◽

Polyherbal Formulation ◽

Ic50 Values

Introduction: The use of natural products as anticancer agents has a long history that began with folklore medicine and through the years has been incorporated into traditional and allopathic medicine. Several drugs currently used are derived from medicinal plants. Objective: The main objective of this study is to investigate the cytotoxic potential of hepatoprotective polyherbal formulation in normal and cancer cell lines. Methods: A 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was utilized to screen the cytotoxic activity. Results: The results revealed that the formulation does not induce much mortality in normal liver and kidney cell lines, and LC50 value of liver cell lines was found 1716.355 μg/ml and kidney cell lines 2464.910 μg/ml. The in vitro anticancer activity was performed on liver, colon, and prostate cancer cell lines, and IC50 values are found 2.077, 3.850, and 11.989 μg/ml, respectively, which show excellent anticancer activity. Conclusion: Based on the results obtained, the hepatoprotective polyherbal formulation is safe for normal cells and cytotoxic for cancer cells. Further, identification and quantification of phytoconstituents responsible for the activity are in progress.

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The Limitations of Collagen/CPP Hybrid Peptides as Carriers for Cancer Drugs to FaDu Cells

Molecules ◽

10.3390/molecules24040676 ◽

2019 ◽

Vol 24 (4) ◽

pp. 676 ◽

Cited By ~ 1

Author(s):

Kevin Ho ◽

Cristobal Morfin ◽

Katarzyna Slowinska

Keyword(s):

Cell Surface ◽

Cell Lines ◽

Cellular Uptake ◽

Hypopharyngeal Carcinoma ◽

Uptake Mechanism ◽

Microscopy Imaging ◽

Fadu Cell ◽

Hybrid Peptides ◽

Fadu Cells

The in vitro efficacy of cancer prodrugs varies significantly between malignant cell lines. The most commonly identified problems relate to delivery: uptake mechanism, endosomal entrapment, and drug release. Here we present the study of collagen/cell penetrating hybrid (COL/CPP) peptide carriers intended to deliver paclitaxel to the hypopharyngeal carcinoma (FaDu) cells. Confocal microscopy imaging revealed the surprising response of FaDu cell to COL/CPP in comparison to previously studied cancer cell lines: hybrid peptides that carry both COL and CPP domain adsorb on the FaDu cell surface. While the CPP domain was design to facilitate the cellular uptake, in the case of FaDu cells, it also induced detrimental interactions with the cell membrane. Despite surface adsorption, the colocalization study with endosomal markers EEA1 and LAMP1 reveals that COL/CPP is internalized via endosomal pathway, peptides are able to escape before lysosome formation and release paclitaxel. Therefore, the main obstacle for paclitaxel delivery to FaDu cells appears to be related to cell surface properties. This behavior seems specific to FaDu cells, and could be linked to previously reported overexpression of T5, heparanase splice variants that produces protein lacking enzymatic activity of heparanase. This results in increased concentration of HSPG on FaDu cell surface, and possibly creates a barrier for cellular uptake of highly charged COL/CPP.

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