A phenotypic approach to probing cellular outcomes using heterobivalent constructs

2020 ◽  
Vol 56 (30) ◽  
pp. 4216-4219
Author(s):  
Rohit Bhadoria ◽  
Kefeng Ping ◽  
Christer Lohk ◽  
Ivar Järving ◽  
Pavel Starkov
Keyword(s):  
Cell Viability ◽  
Cellular Uptake

By comparing the induction of the phenotype of interest, cell viability and cellular uptake, we demonstrate that various conjugates lead to divergent cellular outcomes.

Probing Cellular Outcomes Using Heterobivalent Constructs

2019 ◽  
Author(s):  
Rohit Bhadoria ◽  
Kefeng Ping ◽  
Christer Lohk ◽  
Ivar Järving ◽  
Pavel Starkov
Keyword(s):  
Cell Viability ◽  
Small Molecules ◽  
Cellular Uptake ◽  
Kinase Inhibitors ◽  
Rho Kinase ◽  
Intracellular Delivery ◽  
Rho Kinase Inhibitors

<div> <div> <div> <p>Conjugation techniques are central to improving intracellular delivery of bioactive small molecules. However, tracking and assessing the overall biological outcome of these constructs remains poorly understood. We addressed this issue by having developed a focused library of heterobivalent constructs based on Rho kinase inhibitors to probe various scenarios. By comparing induction of a phenotype of interest vs. cell viability vs. cellular uptake, we demonstrate that such conjugates indeed lead to divergent cellular outcomes. </p> </div> </div> </div>

scholarly journals Cellular uptake of ribonuclease A-functionalised core–shell silica microspheres

2014 ◽  
Vol 2 (42) ◽  
pp. 7307-7315 ◽  
Author(s):  
G. F. Chimonides ◽  
J. M. Behrendt ◽  
E. Chundoo ◽  
C. Bland ◽  
A. V. Hine ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cellular Uptake ◽  
Ribonuclease A ◽  
Core Shell ◽  
Protein Transduction ◽  
Silica Microspheres ◽  
P Protein

Protein transduction: core–shell microspheres have been synthesised and coupled to ribonuclease A. Cellular uptake of these microspheres causes significantly reduced levels of intracellular RNA and reduced cell viability demonstrating that core–shell microsphere-mediated delivery of active enzymes into cells is effective.

scholarly journals Biotin-Decorated PAMAM G4.5 Dendrimer Nanoparticles to Enhance the Delivery, Anti-Proliferative, and Apoptotic Effects of Chemotherapeutic Drug in Cancer Cells

Pharmaceutics ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 443 ◽  
Author(s):  
Endiries Yibru Hanurry ◽  
Tefera Worku Mekonnen ◽  
Abegaz Tizazu Andrgie ◽  
Haile Fentahun Darge ◽  
Yihenew Simegniew Birhan ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Cellular Uptake ◽  
Drug Loading ◽  
In Vitro Cytotoxicity ◽  
Amide Bond ◽  
Solid Cancer

Biotin receptors are overexpressed by various types of solid cancer cells and play a significant role in tumor metabolism, growth, and metastasis. Thus, targeting the biotin receptors on tumor cells may enhance the efficiency and reduce the side-effects of chemotherapy. The aim of this study was to develop a biotin-coupled poly(amido)amine (PAMAM) (PG4.5) dendrimer nanoparticle to enhance the tumor-specific delivery and intracellular uptake of anticancer drugs via receptor-mediated endocytosis. We modified PG4.5 with diethylenetriamine (DETA) followed by biotin via an amide bond and characterized the resulting PG4.5-DETA-biotin nanoparticles by 1H NMR, FTIR, and Raman spectroscopy. Loading and releasing of gemcitabine (GEM) from PG4.5-DETA-biotin were evaluated by UV–Visible spectrophotometry. Cell viability and cellular uptake were examined by MTT assay and flow cytometry to assess the biocompatibility, cellular internalization efficiency and antiproliferative activity of PG4.5-DETA-biotin/GEM. Gemcitabine-loaded PG4.5-DETA-biotin nanoparticles were spherical with a particle size of 81.6 ± 6.08 nm and zeta potential of 0.47 ± 1.25 mV. Maximum drug-loading content and encapsulation efficiency were 10.84 ± 0.16% and 47.01 ± 0.71%, respectively. Nearly 60.54 ± 1.99% and 73.96 ± 1.14% of gemcitabine was released from PG4.5-DETA-biotin/GEM nanoparticles after 48 h at the acidic pH values of 6.5 and 5, respectively. Flow cytometry and fluorescence microscopy of cellular uptake results revealed PG4.5-DETA-biotin/GEM nanoparticles selectively targeted cancer cells in vitro. Cytotoxicity assays demonstrated gemcitabine-loaded PG4.5-DETA-biotin significantly reduced cell viability and induced apoptosis in HeLa cells. Thus, biotin-coupled PG4.5-DETA nanocarrier could provide an effective, targeted drug delivery system and selectively convey gemcitabine into tumor cells.

scholarly journals Cellular Pharmacology of Curcumin With and Without Piperine

2021 ◽  
Author(s):  
Rama I Mahran ◽  
Pan Shu ◽  
Justin Colacino ◽  
Magda M Hagras ◽  
Duxin Sun ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cellular Uptake ◽  
Mcf10a Cell ◽  
Cellular Pharmacology ◽  
Normal Human Breast ◽  
Breast Stem Cell ◽  
Breast Cells ◽  
Normal Human ◽  
Mcf10a Cells ◽  
Mammosphere Assay

Prior reports have suggested that piperine enhances curcumin anti-carcinogenesis. We tested the hypothesis that piperine increases the intracellular concentrations of curcumin by improving intracellular uptake or reducing curcumin efflux or metabolism in breast cells. We incubated SUM149, MCF10A, primary normal human breast cells, ALDH+, and ALDH-CD44+24- SUM149 cells with curcumin with or without piperine at concentrations 1 uM to 15 uM for time periods of 15 minutes to 24 hours. We assayed cell viability by MTT assay and proliferation by primary mammosphere assay. Curcumin and its metabolites were assayed using liquid chromatography mass spectroscopy. Curcumin, but not piperine, showed significantly higher effects on the viability of breast cancer SUM149 cells than in non-tumorigenic MCF10A cells. Curcumin + piperine synergistically reduced viability of SUM149 cells but had a concentration dependent effect upon MCF10A cell viability. Cellular uptake of curcumin in SUM149 is significantly higher, while the efflux in SUM149 is significantly lower than in MCF10A, which correlated with cell viability. Piperine did not alter curcumin cellular uptake, efflux, or metabolism in any of the cell models. The observed synergism of piperine+curcumin in reducing breast stem cell self renewal is likely due to independent anti-carcinogenesis effects rather than any effects upon intracellular curcumin concentrations.

scholarly journals Synergistic effect of cold atmospheric pressure plasma and free or liposomal doxorubicin on melanoma cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Konstantina Pefani-Antimisiari ◽  
Dimitrios K. Athanasopoulos ◽  
Antonia Marazioti ◽  
Kyriakos Sklias ◽  
Maria Rodi ◽  
...  
Keyword(s):  
Synergistic Effect ◽  
Cell Viability ◽  
Cellular Uptake ◽  
Combined Treatment ◽  
Melanoma Cells ◽  
Atmospheric Plasma ◽  
Experimental Conditions ◽  
Trypan Blue Exclusion ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
First Time

AbstractThe aim of the present study was to investigate combined effects of cold atmospheric plasma (CAP) and the chemotherapeutic drug doxorubicin (DOX) on murine and human melanoma cells, and normal cells. In addition to free drug, the combination of CAP with a liposomal drug (DOX-LIP) was also studied for the first time. Thiazolyl blue tetrazolium bromide (MTT) and Trypan Blue exclusion assays were used to evaluate cell viability; the mechanism of cell death was evaluated by flow cytometry. Combined treatment effects on the clonogenic capability of melanoma cells, was also tested with soft agar colony formation assay. Furthermore the effect of CAP on the cellular uptake of DOX or DOX-LIP was examined. Results showed a strong synergistic effect of CAP and DOX or DOX-LIP on selectively decreasing cell viability of melanoma cells. CAP accelerated the apoptotic effect of DOX (or DOX-LIP) and dramatically reduced the aggressiveness of melanoma cells, as the combination treatment significantly decreased their anchorage independent growth. Moreover, CAP did not result in increased cellular uptake of DOX under the present experimental conditions. In conclusion, CAP facilitates DOX cytotoxic effects on melanoma cells, and affects their metastatic potential by reducing their clonogenicity, as shown for the first time.

Probing Cellular Outcomes Using Heterobivalent Constructs

2019 ◽  
Author(s):  
Rohit Bhadoria ◽  
Kefeng Ping ◽  
Christer Lohk ◽  
Ivar Järving ◽  
Pavel Starkov
Keyword(s):  
Cell Viability ◽  
Small Molecules ◽  
Cellular Uptake ◽  
Kinase Inhibitors ◽  
Rho Kinase ◽  
Intracellular Delivery ◽  
Rho Kinase Inhibitors

<div> <div> <div> <p>Conjugation techniques are central to improving intracellular delivery of bioactive small molecules. However, tracking and assessing the overall biological outcome of these constructs remains poorly understood. We addressed this issue by having developed a focused library of heterobivalent constructs based on Rho kinase inhibitors to probe various scenarios. By comparing induction of a phenotype of interest vs. cell viability vs. cellular uptake, we demonstrate that such conjugates indeed lead to divergent cellular outcomes. </p> </div> </div> </div>

scholarly journals Lipidated Peptidomimetic Ligand-Functionalized HER2 Targeted Liposome as Nano-Carrier Designed for Doxorubicin Delivery in Cancer Therapy

2021 ◽  
Vol 14 (3) ◽  
pp. 221
Author(s):  
Himgauri Naik ◽  
Jafrin Jobayer Sonju ◽  
Sitanshu Singh ◽  
Ioulia Chatzistamou ◽  
Leeza Shrestha ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cancer Cells ◽  
Cellular Uptake ◽  
Targeted Delivery ◽  
Chemotherapeutic Agents ◽  
In Vivo Studies ◽  
Breast Cancer Cell Lines ◽  
In Vivo Evaluation ◽  
Lipid Film

The therapeutic index of chemotherapeutic agents can be improved by the use of nano-carrier-mediated chemotherapeutic delivery. Ligand-targeted drug delivery can be used to achieve selective and specific delivery of chemotherapeutic agents to cancer cells. In this study, we prepared a peptidomimetic conjugate (SA-5)-tagged doxorubicin (Dox) incorporated liposome (LP) formulation (SA-5-Dox-LP) to evaluate the targeted delivery potential of SA-5 in human epidermal growth factor receptor-2 (HER2) overexpressed non-small-cell lung cancer (NSCLC) and breast cancer cell lines. The liposome was prepared using thin lipid film hydration and was characterized for particle size, encapsulation efficiency, cell viability, and targeted cellular uptake. In vivo evaluation of the liposomal formulation was performed in a mice model of NSCLC. The cell viability studies revealed that targeted SA-5-Dox-LP showed better antiproliferative activity than non-targeted Dox liposomes (Dox-LP). HER2-targeted liposome delivery showed selective cellular uptake compared to non-targeted liposomes on cancer cells. In vitro drug release studies indicated that Dox was released slowly from the formulations over 24 h, and there was no difference in Dox release between Dox-LP formulation and SA-5-Dox-LP formulation. In vivo studies in an NSCLC model of mice indicated that SA-5-Dox-LP could reduce the lung tumors significantly compared to vehicle control and Dox. In conclusion, this study demonstrated that the SA-5-Dox-LP liposome has the potential to increase therapeutic efficiency and targeted delivery of Dox in HER2 overexpressing cancer.

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