Ongoing projects

Regional Hospital Liberec

„Inhibition of oncogenic microRNA-21 in glioblastoma.“

Annotation: Short regulatory microRNA-21 (miR-21) plays an important role in glioblastoma development. Its increased level is detectable in patients with glioblastoma and contributes to tumor cell survival and disease progression. Using synthetic complementary RNA (antimiR) bound to a suitable carrier (nanoparticles, NPs), we can specifically reduce the level of targeted miR-21. The antimiR-21 / NP system has already been successfully used to reduce miR-21 in colon cancer cells (CT26 cell line) in dr. Benson´s laboratory. That time we used nanodiamond as a carrier, but it is not yet suitable for human application. In the current project, we will primarily use liposomes to determine the effectiveness of antimiR on glioblastoma cells. Subsequently, we plan to use biological nanoparticles of gold, which are biodegradable compared to nanodiamond, and we plan to use them in the subsequent project. Here, the effector antimiR-21 will be electrostatically bound to the carrier, which will ensure its delivery to the cells, release into the cytoplasm and, last but not least, increase the time of antimiR-21 in the tumor microenvironment (important in case of in vivo application in the future). The RNA will be released from the NP due to pH-sensitive binding after internalization of the antimiR-NP complex into the tumor cell. This internalization mostly takes place by endocytosis, and lowering the pH in the endosome is sufficient to release the RNA from the nanocarrier. In this method of antimiR-NP internalization, the endosome is apparently destabilized and the effector RNA enters the cytoplasm, where it binds to the sequence-homologous target miR-21 and the resulting duplex is subsequently degraded. A reduced level of miR-21 leads to impaired tumor cell proliferation.

Grant agency of Charles University

Grant Project: In vitro characterization of electroactive 3D scaffolds and electric stimulation of cells for tissue engineering

Annotation: The project will focus on in vitro testing of 3D nanofiber scaffolds suitable for cell culture. The proposed material should also be electroactive, for example,due to the portion of graphene intercalated between the fibers of the biodegradable polymer. Testing of the cytotoxicity of the material, cell differentiation and subsequently also their electrical stimulation will first be performed on suitable bone and nerve cell lines and afterward with primary cells. The experiments will be performed at first on a suitable cell line SAOS-2, whose cells are derived from osteosarcoma and are widely used in laboratories for their greatly resistance and easy handling. The degree of adhesion of the SAOS-2 cells will be monitored at first. Adhesion is affected by the chemical and physical properties of the material. The material will be exposed to surface modifications (pressing, oxygen plasma treatment) to achieve optimal adhesion properties. Further, the morphology, proliferation, and differentiation of cells will be monitored depending on the interaction with particular materials. The first year of the project will include working with the SAOS-2 cell line and the optimization of the existing methodology with the simultaneous introduction of new protocols for the cultivation and differentiation of the SH-SY5Y cell line. When cultured, SH-SY5Y cells form two phenotypes - neural and epithelial - which are unstable and must be chemically stimulated to maintain the required (neural)

phenotype. After testing the cytotoxicity of the materials by SAOS-2 resistant cell line and after optimizing the protocols for SH-SY5Y cell line, the cultivation of SH-SY5Y cells on the mentioned tested materials will follow, probably for the second year. The next step, probably for the third year of the project, will be to test a laboratory device capable of electrically stimulating the cells on the tested scaffold and to analyze properties of cells. By electrostimulation, we can probably increase the osteo-differentiation of osteoblasts and, for example,

influence the orientation of nerve cells and at the same time create a channel of interconnected neurons using the repair of the damaged nervous system. The aim of our work will be to design and test suitable material for use in tissue engineering, together with partners from TUL, specifically 3D electroactive nanofiber material serving as a scaffold of cells for their possible electrical stimulation and subsequent implantation into damaged nerve or connective tissue. Detailed characterization of cell growth on the prepared material will be followed by back-validation and verification of the importance of specific sample parameters. For this reason, we assume repeated modifications and retesting in cell cultures until the optimal surface for cell growth and stimulation is found.

TAČR SIGMA 2024

Degradable sandwich fibrous hydrogels for moist healing of hard-to-heal wounds

Annotation: Along with higher demands on the quality of patient care and lives, the request for new therapeutic methods is also increasing. The main goal of the project is to develop and optimize products for infected wound healing based on biodegradable submicron fibers from renewable sources. The primary layer of the sandwich membrane will be prepared from hydrophilic polymers modified with bioactive molecules giving the product antiseptic and/or healing properties. The secondary transparent layer will be composed of hydrophobic polymers, which will allow monitoring of wound healing and prevent the wound from drying out. The final demonstration of the healing and antiseptic properties of the membrane will allow the newly developed medical device to be brought closer to clinical practice.