Tissue cultures and Biochemical Engineering

Head of Unit
Dr.rer.nat. Ing. Helena Kanďárová, ERT

Lead scientists
DVM. Štefan Bezek, DSc.

Independent scientists
Ing. Lucia Račková, Ph.D.

Scientists
Ing. Lucia Milec (member of ONTOX/ASPIS)
Mgr. Katarína Pružinská, PhD. (member of ONTOX/ASPIS)
Ing. Klára Gabrišová

Doctoral students
Ing. Peter Pôbiš (supervisor Dr. Kanďárová)
Ing. Sirwan Mahmood (supervisor Dr. Kanďárová)

Graduates
Julia Kubalcová (supervisor Dr. Kanďárová)
Adriena Milana Palenčárová (supervisor Dr. Kanďárová)

Technical staff

Research

• Research into the pathophysiology of ageing and concomitant diseases
• Research into the mechanisms of development and progression of metabolic syndrome
• Investigation into the use of glycobiomarkers in metabolic diseases and aging
• Research into the toxicological mechanisms of medicinal products, new materials, and medical devices
• Research into the therapeutic potential of substances of natural and synthetic origin
  Validation of in vitro methods for use in regulatory toxicology

Experimental models

• Cellular models of inflammation, aging, and oxidative stress
• Cellular models for in vitro toxicity testing of substances
• 3D reconstituted human cell models for testing the safety of chemicals and medical devices
• Animal models of aging and metabolic syndrome
• In vitro inhalation testing – Vitrocell Alpha Cloud system
• Microfluidic platform MIVO(R)

Methods

• In vitro cytotoxicity tests according to ISO and OECD
• Glycomic analysis of blood sera and cell lysates using mass spectrometry
• Flow cytometry, western blot, and immunocytochemistry
• Use of fluorescent probes for evaluation of oxidative stress and toxicological parameters
• ELISA and multiplex bead array test for the assessment of inflammatory and metabolic analytes
• Non-invasive measurement of blood pressure and pulse in experimental animals 
• Isolation of primary cells also using immunomagnetic separation

Cell bank and 3D tissue models

• Skin cells: patient and mouse fibroblasts, immortalized and primary keratinocytes, melanocytes
• Metabolic cells: primary rat hepatocytes, rat insulinoma INS-1E b cells
• Cardiovascular system cells: HUVEC, rat embryonic H9C2 cardiomyocytes
• CNS cells: mouse BV-2 microglia, neuron-like PC12 cells
• Blood cells: mouse RAW264.7 macrophages
• Tumor cells: human cervical cancer cells HeLa, cells of malignant melanoma A375
• MatTek’s patented 3D tissue model systems (EpiDerm, EpiOcular, EpiIntestinal, EpiAirway, EpiOral, EpiGingival)

Projects

International projects

EU-NETVAL International Thyroid Validation Study

Project duration: 01/01/2021 – 01/01/2023

Project leader: Dr.rer.nat., Ing. Helena Kanďárová, ERT

CEM SAS set up the Slovak SK-NETVAL laboratory in 2020 in response to the provision of Directive

2010/63/EU on the protection of animals used for scientific purposes, which requests that

the EU Member States assist the European Commission in the validation of alternative

methods. It is a member of EU-NETVAL, a EURL ECVAM network of specialised

laboratories located in the EU member states.

EU-NETVAL’s mission is to provide support for EURL ECVAM validation studies to assess

the reliability and relevance of alternative methods that have the potential to replace, reduce

or refine the use of animals for scientific purposes. Currently, the network counts 35 EU

members, selected against pre-defined eligibility criteria and endorsed by the National

Contact Points for the Directive 2010/63/EU implementation. Slovak NETVAL laboratory is

working in the GLP mode.

• TRAIN-SAFEMDs – Training Network For Improving Of Safety Of Medical Devices – Focus On Oral Cavity

Project duration: 01/01/2020 – 31/12/2022

Project leader: Dr.rer.nat., Ing. Helena Kanďárová, ERT

§Horizon 2020: ONTOX – Ontology-driven and artificial intelligence-based repeated dose toxicity testing of chemicals for next generation risk assessment

Project duration: 01/05/2021 – 01/05/2026

Project leader: Dr.rer.nat., Ing. Helena Kanďárová, ERT

• COST Action 21139: IMPROVE – 3RS concepts to improve the quality of biomedical science (IMPROVE)

   Project duration: 21/10/2022- 20/10/2026

Project leader: Dr.rer.nat., Ing. Helena Kanďárová, ERT

• COST Action 21108: NETSKINMODELS – European Network for Skin Engineering and Modeling (NETSKINMODELS)

Project duration: 10/2022 –10/2026

Project leader: Dr.rer.nat., Ing. Helena Kanďárová, ERT

National projects since 2008

EU Structural Funds Research and Innovation

• ITMS2014+ 313021Y920 The study of structural changes of complex glycoconjugates in the process of inherited metabolic and civilization diseases

Project duration: 01/03/2021 – 30/06/2023

Project leader: Ing. Zuzana Brnoliaková, Ph.D.

Slovak Research and Development Agency

• APVV-19-0591 In vitro biocompatibility testing of medical devices (MDs) and new generation bio-materials for MDs

Project duration: 01/07/2020 – 30/06/2024

Project leader: Dr.rer.nat., Ing. Helena Kanďárová, ERT

• APVV-18-0336 TOXINOVAGE – Innovative approaches in toxicology of ageing

Project duration: 01/07/2019 – 30/06/2023

Project leader: Ing. Lucia Račková, Ph.D.

Scientific Grant Agency of the Ministry of Education, Science, Research and Sport of the SR

• VEGA 2/0086/08 Developmental Origin of Metabolic Syndrome: Hypertension, Diabetes and Dyslipidemia

Project duration: 01/01/2008 – 31/12/2010

Project leader: DVM. Štefan Bezek, DSc.

• VEGA 2/0104/21 The use of mass spectrometry for comparative study of different rats strains glycoprofiles within metabolic disturbances intervention

Project duration: 01/01/2021 – 31/12/2023

Project leader: Ing. Zuzana Brnoliaková, Ph.D.

• VEGA 2/0056/09 Could the diabetic cataract prevention be achieved by substituted pyridoindoles with their antioxidant and aldose reductase inhibitory potential? Study on in vitro cultivated eye lenses from rats.

Project duration: 01/01/2009 – 31/12/2011

Project leader: Ing. Zuzana Brnoliaková, Ph.D.

• VEGA 2/0153/20 Bio-compatibility assessment of medical devices and novel medical device materials using in vitro methods based on 3d reconstructed human tissue models.

Project duration: 01/01/2020 – 31/12/2023

Project leader: Dr.rer.nat., Ing. Kanďárová Helena, ERT

• VEGA 2/0041/17 Redox Homeostasis, Proteostasis, and Inflammation as Potential Targets For Influencing Ageing and Age-Related Diseases: Modulation by the compounds of natural and synthetic origin. Project duration: 01/01/2017 – 31/12/2020

Project leader: Ing. Lucia Račková, Ph.D.

• VEGA 2/0031/12 Brain Aging and Neuroprotective Antioxidants: Influence of Glia as Therapeutic Strategy? Project duration: 01/01/2012 – 31/12/2015

Project leader: Ing. Lucia Račková, Ph.D.

University pedagogy

Lectures
Subject: Cellular and tissue cultures
Faculty of Chemical and Food Technology STU
Lecturer: Dr.rer.nat., Ing. Kanďárová Helena, ERT

Subject: Biomolecular design
Faculty of Chemical and Food Technology STU
Lecturer: Ing. Lucia Račková, Ph.D.

Laboratory classes

Subject: Cellular and tissue cultures
Faculty of Chemical and Food Technology STU
Instructor: Dr.rer.nat., Ing. Kanďárová Helena, ERT

Ing. Peter Pôbiš

Theses

Dissertations
Sonam KAPOOR. Neonatal asphyxia: changes in glucose metabolism in newborn

rats. Supervisor Dr. Brnoliaková, Bratislava, 2022.

• Hande ÖZBAȘAK (1st y.). Toxicology of novel molecular sensors and NO donors and their application in cellular models. Supervisor: Dr. Račková

• Peter PÔBIŠ (1st y.). In Vitro 3D reconstitution models of human tissues – use and implementation in toxicology as a substitute for in vivo tests. Supervisor: Dr. Kanďárová

• Erika CSEKES. Protoapigenone 1′-O-Butyl ether as a small senogenous molecule in antitumor therapy. Supervisor: Dr. Račková, Bratislava, 2020. 139 s.

• Martin ŠKANDÍK. Cellular and molecular mechanisms of ageing and related diseases: Modulation with substances of synthetic and natural origin. Supervisor: Dr. Račková, Bratislava, 2020. 157 s.

• Nataša MRVOVÁ. General pathogenic mechanisms in age-related neurodegeneration: modulation with natural and synthetic substances. Supervisor: Dr. Račková, Bratislava, 2016. 129 s.

Diploma theses

• Peter PÔBIŠ. Reconstituted model of 3D cornea for the study of phototoxicity of selected drugs and medical devices. FCHPT STU, Bratislava 2021. Supervisor: Dr. Kanďárová

• Martin ŠKANDÍK. Novel quinone derivative of quercetin: Cytotoxicity and anti-inflammatory potential in the neuroglia cell model. FCHPT STU, Bratislava 2016. Supervisor: Dr. Račková

Bachelor theses

• Júlia KUBALCOVÁ. FCHPT STU, Bratislava 2022. New approaches for assessing the biocompatibility of medical devices: Cellular lines and 3D models. Supervisor: Dr. Kanďárová, consultant: Ing. Pôbiš

Selected publications

RAČKOVÁ, L.**, CSEKES, E. Redox aspects of cytotoxicity and anti-neuroinflammatory profile of chloroquine and hydroxychloroquine in serum-starved BV-2 microglia. In Toxicology and Applied Pharmacology, 2022, 447:116084. (2021: 4.46 – IF, Q2 – JCR, Q1 – SJR, current contents – CCC).

KANĎÁROVÁ, H.**, HAYDEN, P.J. Standardised Reconstructed Skin Models in Toxicology and Pharmacology: State of the Art and Future Development. In Handbook of Experimental Pharmacology. – Springer Nature, 2021, 2021, vol. 265, p. 57-71. (2020: 1.605 – SJR, Q1 – SJR). ISSN 0171-2004.

KANĎÁROVÁ, H. Alternative methods to animal experimentation and their role in modern toxicology. In Toxicological Risk Assessment and Multi-System Health Impacts from Exposure. – Elsevier Inc., Academic Press, 2021, part 1, chapter 2, P. 13-22. ISBN 978-0-323-85215-9.

SPIELMANN, H.**, KANĎÁROVÁ, H. Integration of Advanced Technologies into Regulatory Toxicology. In Regulatory Toxicology. 2nd edition. – Cham: Springer Nature Switzerland AG, 2021, p. 149-161. ISBN 978-3-030-57498-7.

SVOBODOVÁ, L.**, RUCKI, M., VLKOVÁ, A., KEJLOVÁ, K., JÍROVÁ, D., DVOŘÁKOVÁ, M., KOLÁŘOVÁ, H., KANĎÁROVÁ, H., PÔBIŠ, P., HEINONEN, T., MALÝ, M. Sensitization potential of medical devices detected by in vitro and in vivo methods. In ALTEX : Alternatives to Animal Experimentation, 2021, vol. 38, no. 3, p. 419-430. (2020: 6.043 – IF, Q1 – JCR, 0.975 – SJR, Q1 – SJR). ISSN 1868-596X.

VINKEN, M., BENFENATI, E., BUSQUET, F., CASTELL, J., CLEVERT, D.-A., DE KOK, T. M.,  DIRVEN, H., FRITSCHE, E., GERIS, L., GOZALBES, R., HARTUNG, T., JENNEN, D., JOVER, R., KANĎÁROVÁ, H., KRAMER, N., KRUL, C., LEUCHTEFELD, T., MASEREEUW, R., ROGGEN, E., SCHALLER, S., VANHAECKE, T., YANG, Ch., PIERSMA, A.H. Safer chemicals using less animals: kick-off of the European ONTOX project. In Toxicology, 2021, vol. 458, art. no. 152846. (2020: 4.221 – IF, Q2 – JCR, 1.067 – SJR, Q1 – SJR current contents – CCC). (2021 – Current Contents). ISSN 0300-483X. (H2020, ONTOX : Ontology-driven and artificial intelligence-based repeated dose toxicity testing of chemicals for next generation risk assessment)

RAČKOVÁ, L.**, MACH, M. BRNOLIAKOVÁ, Z. An update in toxicology of ageing. In Environmental Toxicology and Pharmacology, 2021, vol. 84, art. no. 103611. (2020: 4.860 – IF, Q1 – JCR, 0.924 – SJR, Q1 – SJR, current contents – CCC). (2021 – Current Contents). ISSN 1382-6689.

ŠKANDÍK, M., MRVOVÁ, N., BEZEK, Š., RAČKOVÁ, L**. Semisynthetic quercetin-quinone mitigates BV-2 microglia activation through modulation of the Nrf2 pathway. In Free Radical Biology and Medicine, 2020, vol. 152, p. 18-32. (2019: 6.170 – IF, Q1 – JCR, 1.841 – SJR, Q1 – SJR, current contents – CCC). (2020 – Current Contents). ISSN 0891-5849. (Dissertation of M.Š.)

CSEKES, E., VÁGVÖLGYI, M., HUNYADI, A., RAČKOVÁ, L**. Protoflavones in melanoma therapy: Prooxidant and pro-senescence effect of protoapigenone and its synthetic alkyl derivative in A375 cells. In Life Sciences, 2020, vol. 260, art. no. 118419. (2019: 3.647 – IF, Q2 – JCR, 1.031 – SJR, Q1 – SJR, current contents – CCC). (2020 – Current Contents). ISSN 0024-3205. (Dissertation of E.C.)  

MRVOVÁ, N., ŠKANDÍK, M., BEZEK, Š., RAČKOVÁ, L**. Protective effect of semisynthetic and natural flavonoid on aged rat microglia-enriched cultures. In Neurotoxicity Research, 2019, vol. 36, no. 4, p. 844-858. (2018: 3.311 – IF, Q2 – JCR, 0.941 – SJR, Q1 – SJR). ISSN 1029-8428. (Dissertation of N.M.)

MRVOVÁ, N., ŠKANDÍK, M., BEZEK, Š., SEDLÁČKOVÁ, N., MACH, M., GÁSPÁROVÁ, Z., LUPTÁKOVÁ, D., PADEJ, I., RAČKOVÁ, L**. Pyridoindole SMe1EC2 as cognition enhancer in ageing-related cognitive decline. In Interdisciplinary Toxicology, 2017, vol. 10, no. 1, p. 11-19. (2016: 0.409 – SJR, Q3 – SJR). (2017 – SCOPUS). ISSN 1337-6853.

MRVOVÁ, N., ŠKANDÍK, M., KUNIAKOVÁ, M., RAČKOVÁ, L**. Modulation of BV-2 microglia functions by novel quercetin pivaloyl ester. In Neurochemistry International, 2015, vol. 90, p. 246-254. (2014: 3.092 – IF, Q2 – JCR, 1.371 – SJR, Q2 – SJR, current contents – CCC). (2015 – Current Contents). ISSN 0197-0186. (Dissertation of N.M.)