Karina Kwapiszewska

Polish Academy of Sciences | PAN · Institute of Physical Chemistry

Bevacizumab is a biological drug that is now extensively studied in clinics against various types of cancer. Although bevacizumab's action is preferably extracellular, there are reports suggesting its internalization into cancer cells, consequently decreasing its therapeutic potential. Here we are solving this issue by applying fluorescence correla...

Fluorescent crystals of bisantrene were detected in water-based buffers. A similar size range of crystals and cells prevents its uptake for ATP detection.

Biocompatible polyacrylamide gel and core-shell nanoparticles (NPs) were synthesized using a one-step electrochemically initiated gelation. Constant-potential electrochemical decomposing of ammonium persulfate initiated the copolymerization of N-isopropyl acrylamide, methacrylic acid, and N,N'-methylenebisacrylamide monomers. This decomposing poten...

Hypothesis Most experimental procedures applied in modern biology involve cargo delivering into cells. One of the ways to cargo introduction is osmotic-mediated intracellular vesicle swelling. However, its widespread use was hindered due to cargo size (<10 nm) and cell-type-related restrictions. We addressed the issue of the composition of colloida...

Nitrophenols (NPs) are hazardous pollutants found in various environmental matrices, including ambient fine particulate matter (PM2.5), agricultural residues, rainwater, wildfires, and industrial wastes. This study showed for the first time the effect of three pure nitrophenols and their mixture on human lung cells to provide basic understanding of...

Quantitative description of biochemical processes inside living cells and at single-molecule levels remains a challenge at the forefront of modern instrumentation and spectroscopy. This paper demonstrates such single-cell, single-molecule analyses performed to study the mechanism of action of olaparib - an up-to-date, FDA-approved drug for germline...

Targeting cap-dependent translation initiation is one of the experimental approaches that could lead to the development of novel anti-cancer therapies. Synthetic dinucleoside 5',5'-triphosphates cap analogs are potent antagonists of eukaryotic translation initiation factor 4E (eIF4E) in vitro and could counteract elevated levels of eIF4E in cancer...

Biomass burning results in the emission of 10-15 Tg yr-1 of organic carbon (OC) into the atmosphere globally. The UV-light absorbing OC component contains a group of poorly characterized nitrated aromatic compounds (NACs). These NACs are important secondary organic aerosol (SOA) products of urban biomass burning aerosol (BBA) involving nitrogen oxi...

Secondary organic aerosol (SOA) is a major component of airborne fine particulate matter (PM2.5) that contributes to adverse human health effects upon inhalation. Atmospheric ozonolysis of α-pinene, an abundantly emitted monoterpene from terrestrial vegetation, leads to significant global SOA formation; however, its impact on pulmonary pathophysiol...

Understanding the mobility of nano-objects in the eukaryotic cell nucleus, at multiple length-scales, is essential for dissecting nuclear structure-function relationships both in space and in time. Here, we demonstrate, using single-molecule fluorescent correlation spectroscopies, that motion of inert probes (proteins, polymers, or nanoparticles) w...

Wildfires, biofuel combustion, and wood burning emit 10-15 Tg/yr of organic carbon (OC) into the atmosphere globally. The UV-light absorbing component of OC termed as “brown carbon” (BrC) contains a group of poorly characterized nitrated aromatic compounds (NACs). These NACs are important secondary products of urban biomass burning aerosol (BBA) in...

The efficient delivery of drugs to cells depends on their diffusion through the extracellular matrix (ECM) of tissues. Here we present a study of diffusion of nanoprobes of radius from 1 nm to over 100 nm in the ECM of spheroids of three cell types (HeLa, MCF-7 and fibroblasts). We quantified nanoparticle transport in spheroids’ proliferating zone....

Metabolic reactions in living cells are limited by diffusion of reagents in cytoplasm. Any attempt to quantify the kinetics of biochemical reactions in cytosol should be preceded with careful measurements of the physical properties of the cellular interior. Cytoplasm is a complex, crowded fluid characterized by effective viscosity dependent on its...

Nanoviscosity of the cytoplasm is a key factor affecting diffusion of biomolecules and – as a consequence – rates of biochemical reactions in a cell. Nanoviscosity is an outcome of variable chemical and structural factors, which can temporarily change with cell-cycle associated changes of intracellular architecture. Thus, the question arises, wheth...

F1-4393073-Abstract-EAC2019-Faria_Khan

Biochemistry in living cells is an emerging field of science. Current quantitative bioassays are performed ex vivo, thus equilibrium constants and reaction rates of reactions occurring in human cells are still unknown. To address this issue, we present a non-invasive method to quantitatively characterize interactions (equilibrium constants, KD) dir...

Multicellular spheroid is a three-dimensional (3D) cell culture model that mimics cancer tumor environment. Its widespread use for anticancer therapy evaluation is nowadays limited by accessibility of 3D compatible assays. Here, a microfluidic system for spheroid formation, culture and analysis is presented. The system is compatible with standard m...

This work, based on in vivo and in vitro measurements, as well as in silico simulations, provides a consistent analysis of diffusion of polydisperse nanoparticles in cytoplasm of living cells. Using the example of fluorescence correlation spectroscopy (FCS) we show the effect of polydispersity of probes on the experimental results. Although individ...

Cell-on-a-chip systems have become promising devices to study the effectiveness of new anticancer drugs recently. Several microdevices for liver cancer culture and evaluation of the drug cytotoxicity have been reported. However, there are still no proven reports about high-throughput and simple methods for the evaluation of drug cytotoxicity on liv...

The paper describes a cheap and accessible technique of a poly(dimethylsiloxane) (PDMS) master treatment by thermal aging as a step of double casting microfabrication process. Three-dimensional PDMS microstructures could have been achieved using this technique. It was proved, that thermal aging changes nanotopology of a PDMS surface and thus enhanc...

Interest in the microfluidic environment, owing to its unique physical properties, is increasing in much innovative chemical, biological, or medicinal research. The possibility of exploiting and using new phenomena makes the microscale a powerful tool to improve currently used macroscopic methods and approaches. Previously, we reported that an incr...

The development of novel cellular models that can replace animals in preclinical trials of drug candidates is one of the major goals of cell engineering. Current in vitro screening methods hardly correspond with the in vivo situation, whereas there is a lack of assays for more accurate cell culture models. Therefore, development of automated assays...

Early, accurate diagnosis of lysosomal storage disorders is a major challenge, even for trained specialists. Finding innovative, accurate diagnostic methods, and high throughput, cost-effective tools are crucial to medical progress and will contribute to improved quality of life. The goal of this work was to improve currently used protocols to dete...

One of the major advantage of microfluidic systems is a possibility of precise control over fluid flow. However, this can be significantly disturbed by air bubbles penetrating microchannels. Many efforts are taken to improve tubing connections and tightness of experimental set-ups. However, application of gas permeable materials, such as PDMS, whic...

Development of novel cellular models that can replace animals in preclinical trials of drug candidates is one of the major goals of cell engineering. Current in vitro screening methods hardly correspond with the in vivo situation, whereas there is lack of assays for more accurate cell culture models. Therefore, development of automated assays for 3...

a b s t r a c t In this paper, we present a low-cost three-dimensional microfluidic system made of poly(dimethylsiloxane) for tumor spheroid formation and cultivation. Tumor spheroids reaching diameters of up to 300 m were cultured in perfusion microchambers providing extremely low shear stress. Spheroids were cultured for 15 days and high viabilit...

In this work, we present a microfluidic array of microwells for long-term tumor spheroid cultivation and anticancer drug activity evaluation. The three-dimensional microfluidic system was obtained by double casting of poly(dimethylsiloxane). Spheroids of HT-29 human carcinoma cells were cultured in the microsystem for four weeks. After two weeks of...

In the work discussed in this paper, the effect of a high surface-to-volume ratio of a microfluidic detection cell on fluorescence quenching was studied. It was found that modification of the geometry of a microchannel can provide a wider linear range. This is a phenomenon which should be taken into consideration when microfluidic systems with fluo...

In this work, an integrated microfluidic system for biochemical diagnostics of lysosomal storage disorders is presented. The polymeric microsystem consists of a zone for hydrodynamic focusing of cell suspension, a mixing microchannel, and an optical detection module. The system was used for determination of the activity of enzymes deficient in Fabr...

This paper describes a novel method of poly(dimethylsiloxane) double casting utilizing surface modification by thermal aging. The composition of the polymer was optimized and the conditions of thermal aging were set to achieve correct replica-tion of over 98% of microstructures. The microfluidic device fabricated using developed technique was succe...

This paper describes an integrated microfluidic system for intracellular components analysis. The polymeric microdevice consists of a zone for hydrodynamic focusing of cell suspension, a mixing microchannel, and an optical detection module. In this work the system was used to determine the activity of α-galactosidase A (an enzyme deficient in Fabry...

In this paper, a microfluidic system for long-term multicellular tumor spheroids culture and analysis is presented. Human car-cinoma cells were cultured in an array of microwells for four weeks and a slowdown of the proliferation with maintained viability was observed. The construction of the device enabled observation of decreasing spheroid diamet...

We have investigated the kinetics of α-galactosidase A and β-glucocerebrosidase deficient in Fabry and Gaucher diseases, respectively. We have performed spectrofluorymetric measurements of the activity of enzymes using a derivative of 4-methylumbelliferone as a substrate and a human T-cell line as a source of enzymes. We have observed the substrate...

An integrated microfluidic system that coupled lysis of two cell lines: L929 fibroblasts and A549 epithelial cells, with fluorescence-based enzyme assay was developed to determine β-glucocerebrosidase activity. The microdevice fabricated in poly(dimethylsiloxane) consists of three main parts: a chemical cell lysis zone based on the sheath flow geom...

One of the major branches of microfluidic development is cell engineering. A number of devices for cell cultivation, lysis, single-cell analysis and cell-based toxicity tests have been reported in the literature. The variety of structures that can be created leads to devices more closely mimicking the in vivo environment than classic cell cultures....

One of the main scopes of modern cell engineering is development of cellular models that can replace animals in drug screening and toxicological tests, so called alternative methods. Construction of the alternative model is avery challenging task due to arichness of factors creating the in vivo environment. Th e monolayer cell culture — cultivation...

Microfl uidic devices, such as lab-on-a-chip systems, are highly advantageous for cell engineering and cell based assays. It is aparticularly useful approach for development of the in vitro cellular systems mimicking the in vivo environment. In this paper, anovel lab-on-a-chip device for three-dimensional human cell culture and anticancer drug test...

In this paper, a microfluidic system for long-term multicellular tumor spheroids culture and analysis is presented. Human carcinoma cells were cultured in an array of microwells for four weeks and a slowdown of the proliferation with maintained viability was observed. The construction of the device enabled observation of decreasing spheroid diamete...

In this paper, hybrid (PDMS/glass) microfluidic cell culture system (MCCS) integrated with the concentration gradient generator (CGG) is presented. PDMS gas permeability enabled cells’ respiration in the fabricated microdevices and excellent glass hydrophilicity allowed successful cells’ seeding. The human lung carcinoma cells (A549) were cultured...

Miniaturowy system analityczny do hodowli komórkowych Streszczenie. Celem badań było wykonanie miniaturowego systemu przeznaczonego do hodowli komórek adherentnych, a następnie ocena jego przydatności w prowadzeniu analiz z wykorzystaniem hodowanych komórek. W wykonanym mikrosystemie przeprowadzono kilkudniową hodowlę komórek: A549 – komórki nowotw...

The aim of our study was to obtain a microsystem for adherent cell culture and then assess its usefulness for tests on the cultured cells. The culture of the human lung carcinoma cells (A549) and colorectal carcinoma cells (HT-29) was carried on in the microdevice for several days. The microdevice before cell docking in microchambers was especially...

This article presents an overview of various miniaturized devices and technologies developed by our group. Innovative, fast and cheap procedures for the fabrication of laboratory microsystems based on commercially available materials are reported and compared with well-established microfabrication techniques. The modules fabricated and tested in ou...