Home
Biomaterials

Ongoing project NanoSafety and NanoMedicine

Main content

Microfluidic platform for real-life exposure to nanomaterials

The microscopy setup consists of a BioStation IM (Nikon) with automated XYZ stage, temperature and CO2 control for incubation of living cells. Impedance is monitored in real-time with a LCR meter (HIOKI). The platform is equipped with a OB1 MK4 pressure flow controller (Elveflow). Microfluidic chips and Au-microelectrode arrays are designed and made in-house at the clean-room facilities of the Dept. of Physics and Technology (UiB).

Microfluidic platform for label-free, real-time electrical impedance analysis and live-cell imaging of cells exposed to nanomaterials.
Photo:
Ivan Rios Mondragon

Multiplexed microfluidic platform for label-free, real-time electrical impedance spectroscopy (EIS) and live-cell imaging of cells exposed to nanomaterials.

The microscopy setup consists of a fluorescence microscope (Olympus) with automated XYZ stage (Prior Scientific), Andor’s differential spinning disk (DSD) unit with CMOS camera for semi-confocal image acquisition (Oxford Instruments). The microscope is enclosed in an environmental chamber with temperature, humidity and CO2/O2 control for incubation of living cells. EIS instrumentation consists of an Autolab PGSTAT potentiostat (Metrohm) with multiplexing module (up to 16 channels) or 64-channel ISX3 impedance analyser (Sciospec). The platform is equipped with pulsation-free syringe pumps (CETONI), high-precision peristaltic pumps (CETONI) and OB1 MK4 pressure flow controller (Elveflow). Microfluidic chips and Au-microelectrode arrays are designed and made in-house at the clean-room facilities of the Dept. of Physics and Technology (UiB).

Multiplexed microfluidic platform for label-free, real-time electrical impedance spectroscopy (EIS) and live-cell imaging of cells exposed to nanomaterials.
Photo:
Ivan Rios Mondragon

Microfluidic set-up.

Microfluidic set-up. A) Schematic diagram depicting the components of the microfluidic platform. B) Different designs of interdigitated MEAs used. C) Multiplexed microfluidic chip made in polydimethylsiloxane with Au-microelectrode arrays.

Photo:
Ivan Rios Mondragon

 

A549 lung cancer cells exposed to multi-layered graphene encapsulated magnetic nanoparticles

A549 lung cancer cells exposed to multi-layered graphene encapsulated magnetic nanoparticles (GEMNS) for 2h at a flow rate of 1ul/min. A) Impedimetric analysis at 25KHz AC frequency. B) Fold-change relative to control 2 and 22h after treatment. C) Brightfield images 22h after exposure.

Photo:
Ivan Rios Mondragon

Researcher: Ivan Rios-Mondragon, PhD, PI: Mihaela Roxana Cimpan, Professor

 

3D biological models

Advanced in-vitro models to study the interaction of nanomaterials with biological systems

We develop organ-on-a-chip models to understand how nanomaterials interact with biological systems. We develop symplistic, but reliable models to limit/replace the use of animals for testing of nanomaterials present in consumer products and medical devices including nanopharmaceuticals.

Microvasculature on-a-chip model.

Microvasculature on-a-chip model. A) Schematic of the 3D vascular model. Microvascular networks are formed when endothelial cells are co-cultured with fibroblast within a fibrin gel inside the microfluidic chip made of PDMS. The lumen of the blood vessels is accessible for delivery of NMs. B) Microfabricated Si-mold and its replica in PDMS containing the microfluidic network. C) Brightfield micrograph showing the formation of vascular networks of HULEC-5a co-cultured for 7 days with primary lung fibroblasts. D) Immunostained co-cultures. HULEC-5a formed tubular networks.

Photo:
Ivan Rios Mondragon

Breast cancer-on-a-chip model for testing of nanomaterials

Breast cancer-on-a-chip model for testing of nanomaterials
Photo:
Ivan Rios Mondragon

Microfluidic chip for breast cancer-on-a-chip model. Cartoon depicting the different components and cells integrating the breast cancer-on-a-chip model (top-left). Microfluidic chip (bottom-left). 3D reconstruction of HUVEC microvasculature and adjacent MFC10a cancer cell monolayter in the breast cancer chip (right). Empty liposome (Lip-SiR) were perfused in the HUVEC channel for 24 hr. For both cell lines the nucleus is shown in blue (DAPI), membrane in magenta (WGA-AF488) and empty liposome in green (Lip-SiR). Top, merged channels DAPI/WGA-AF488/Lip-SiR. Middle, merged channels DAPI/Lip-SiR. Bottom, single channel for Lip-SiR. White arrows indicate Lip-SiR signal.

Lung-on-a-chip model for testing of nanomaterials

Lung-on-a-chip model for testing of nanomaterials

A) master moulds and replicas in polydimethylsiloxane (PDMS) of top (air) and bottom (liquid) chambers that compose the lung-on-a-chip model. Master moulds were fabricated using photolithography. B) Side view (top) and magnification (bottom) of the fully assembled chip. The air and liquid chambers are interfaced by a thin (20 um) PET microporous membrane comprising 3 um pores.

Photo:
Ivan Rios Mondragon

Researcher: Ivan Rios-Mondragon, PhD, PI: Mihaela Roxana Cimpan, Professor

Label-free impedance-based and electrochemical methods

 

Impedance-based high throughput nanotoxicity screening on adherent and single cells.

Label-free: Impedance based biosensor technology does not require markers or dyes.  Interference-free from Nanomaterials and Nanoparticles perturbation.

xCELLigence system (Agylent Technologies, Inc., USA)

Picture of xCELLigence instrument
Photo:
Photo: Ivan Rios-Mondragón

 

Real-time kinetic readouts: Obtains data continuously from seconds to days.  Simultaneously monitor up to three plates, without scheduling conflicts. Easy workflow: Plate cells, expose to Nanomaterials and begin monitoring.

Measures cell number, size, morphology, and attachment properties, with the ability to perform kinetic analysis of cell invasion/migration (CIM).

AmphaZ30 (Amphasys AG, Lucerne, Switzerland)

Picture of Instrument Ampha Z30
Photo:
Ivan Rios-Mondragón

Amphaz30 is an impedance flow cytometer for high-throughput single-cell characterization without optical components. Single cells pass through a micrometer-sized channel in a chip equipped with microelectrodes. The electrical impedance changes when a cell passes through the applied alternating current (AC) field, permitting cell detection and impedance measurement. The measured impedance is used to assess cellular size, membrane capacitance, and cytoplasm resistance and to differentiate between live and cell cells and modes of cell death.

Cyclic voltammetry for oxidative stress testing.

Photo of Cyclic voltammetry to study nanomaterial-mediated oxidative stress
Photo:
Ivan Rios-Mondragón

We use cyclic voltammetry (CV) to study if nanomaterials may cause oxidative stress in biological systems. The total antioxidant capacity (TAC) of a biofluid reflects the amount of antioxidants available to counteract oxidative stress.

Module for measurement of transepithelial/endothelial electrical resistance (TEER)

Module for measurement of transepithelial/endothelial electrical resistance (TEER)

Top, TEER module placed on top of the lung-on-a-chip and hook up to the wires at the microscope stage. Bottom, magnification showing the gold-plated electrodes that are immersed in the cell culture medium at the inlets/outlets of each cell culture chamber of the lung-on-a-chip

Photo:
Ivan Rios Mondragon

Researcher: Ivan Rios-Mondragon, PhD, PI: Mihaela Roxana Cimpan, Professor

 

HORIZON2020 EU Project "RiskGone"

RiskGONE PROJECT – Science-based Risk Governance of Nano-Technology (nilu.no)

WP5 co-lead and PI: Mihaela Roxana Cimpan, Professor; Researcher: Ivan Rios-Mondragon, PhD; Ying Xue, PhD; Håkon Van Ta, MSc Nano; Arnaud Lemelle, PhD; Anne Marthe Drønen, BSc

NANO2021 RCN project "NanoBioreal"


A sound scientific basis is needed to assess the risks to workers and consumers, to inform regulatory bodies and to ensure a responsible development of nanotechnology. Most of the existing laboratory (in vitro) biological models, exposure systems and doses, as well data (in silico) models do not reflect the real-life exposure to nanomaterials (NMs). A significant source for unreliable results is represented by possible interactions of NMs with the reagents and detection systems for toxicity evaluation. 


The fast pace at which NMs enter the market requires a shift from expensive and ethically doubtful animal testing to innovative, reliable and socially acceptable in vitro and in silico test systems. The aim of NanoBioReal is to establish methods and biological models that reflect real-life exposure and which provide a reliable, robust and efficient platform to evaluate the effects of NMs on human health. Our testing system covers a wide area of biological models, from single cells to three-dimensional (3D) models that simulate tissues and organs. These include air-liquid interface (ALI) models for lung exposure, blood and «organ-on-a-chip» systems (lung and microvasculature-on-a-chip) that measure effects in real-time and can detect relevant effects after both short and longtime exposure. 


To avoid interferences caused by NMs, we established in this project label-free impedance-based methods to evaluate cytotoxicity and cyclic voltammetry to assess oxidative stress caused by NMs. A set of representative NMs has been produced and physico-chemically characterized. Their in vitro cyto- and geno- toxicity was assessed in 2D cellular models and the results were compared to those obtained using the advanced 3D models and an animal model. Advanced models for lung, vasculature and whole blood exposure have been established and used for toxicity, inflammation, oxidative stress, and barrier integrity testing. A microfluidic setup for label-free live monitoring of cells and 3D biological models was established, optimized and its throughput has been increased. The results obtained in the NanoBioReal project deliver reliable, robust and relevant biological and in silico-models to support a “safe(r)-by-design” approach to the development of NMs and to address the needs of various stakeholders and regulators. 


National partners:  Coordinator - Dept. of Clinical Dentistry (IKO), Fac. of Medicine, Univ. of Bergen (UiB); Co-coordinator - National Inst. of Occupational Health (STAMI); the Norwegian Institute for Air Research (NILU); Centre of Molecular Inflammation Research (CEMIR) at the Norwegian University of Science and Technology (NTNU). Subcontractors: NorGenotech. International partners: Catalan Inst. of Nanoscience and Nanotechnology (ICN2) and the University of Gdansk. Collaborators: Dept. of Physics and Technology - NanoPhysics, Bodil Holst, Professor, Martin Greve, Associate Professor (UiB); Dept. of Electrical Engineering (HVL) - Emil Cimpan, Associate Professor; NIOM; TkVest; TkØst.

 

Workshop “New Approach Methodologies for NanoSafety”  and the “NanoBioReal” Project meeting were held in Bergen on March 30th and March 31st, 2023.  The workshop and project meeting were organized by Mihaela R. Cimpan, Ivan Rios-Mondragon, Barbara Pekala, Sigrid Nævdal, the Department of Clinical Dentistry, Faculty of Medicine, University of Bergen.

Coordinator: Mihaela Roxana Cimpan, Professor; Researcher: Ivan Rios-Mondragon, PhD; Ole Bendik Hofshagen, MSc Nano; Marianne Stokka MSc Nano; Medusja Sritharan Nalliah MSC Nano

Research School for Training the Next Generation of Micro- and Nanotechnology Researchers in Norway (TNNN)

EEA Project "TEPCAN"

Theranostic Exosomes in Personalized Cancer Nanomedicine | Warszawski Uniwersytet Medyczny (wum.edu.pl)

WP leader, PI: Mihaela Roxana Cimpan, Professor; Researcher: Ivan Rios-Mondragon, PhD

 

ANR project "NanoMilk"

Metal nanoparticles contamination of milk: mother-to-young transfer and role of extracellular vesicles | ANR

WP leader, PI: Mihaela Roxana Cimpan, Professor; Coordinator: Anne Burtey, PhD (INRAE, Paris); Researcher: Ivan Rios-Mondragon, PhD; Marie Simon, PhD cand (INRAE, Paris).

Use of nanomaterials: Perception and hazard

https://www.uib.no/en/rg/oralhealth/152931/use-nanomaterials-perception-...

PI: Anne Nordrehaug Åstrøm, Professor; Victoria Xenaki, PhD cand.; Mihaela R.Cimpan, Professor Mihaela Cuida Marthinussen, Associate Professor; Daniela E. Costea, Professor; Stein Atle Lie, Professor.

UH-nett Vest Project: New methods for assessing the effects of nanomaterials, MRI and various stressors on cells

https://uhnettvest.no/finner-nye-metoder-for-a-kartlegge-biologiske-effe...

Mihaela Roxana Cimpan, Ivan Rios-Mondragon, Kamal Mustafa (IKO), Emil Cimpan, Alvhild Alette Bjørkum (HVL), Bodil Holst, Martin Greve (IFT), Beate Kluge og Karen Rosendahl (Haukeland University Hospital)

Effects of Artificial Saliva on Silica nanoparticles

Students: Ida Marlene Nygård and Caroline Skjennum

Supervisors: Asgeir Bårdsen, Professor;  Mihaela R. Cimpan, Professor

Senior engineer: Hanzhen Wen

 

Effects of Nanodiamond particles

Nanodiamond particles and stem cells.

Nanodiamond particles: Osteogenicity and safety aspects

Mohammed A. Yassin, Associate Professor; Mohammed Ibrahim, PhD; Mihaela R.Cimpan, Professor, Kamal Mustafa, Professor, Julia Schoelermann, PhD.