Browsing by Author "Patetko, Liene"
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Item Cancer microcell initiation and determination(2021-12) Simsone, Zane; Freivalds, Tālivaldis; Bēma, Dina; Miķelsone, Indra; Patetko, Liene; Bērziņš, Juris; Harju, Līga; Buiķis, Indulis; Department of Human Physiology and BiochemistryBackground: Cancer remains one of the leading causes of death worldwide, despite the possibilities to detect early onset of the most common cancer types. The search for the optimal therapy is complicated by the cancer diversity within tumors and the unsynchronized development of cancerous cells. Therefore, it is necessary to characterize cancer cell populations after treatment has been applied, because cancer recurrence is not rare. In our research, we concentrated on small cancer cell subpopulation (microcells) that has a potential to be cancer resistance source. Previously made experiments has shown that these cells in small numbers form in specific circumstances after anticancer treatment. Methods: In experiments described in this research, the anticancer agents’ paclitaxel and doxorubicin were used to stimulate the induction of microcells in fibroblast, cervix adenocarcinoma, and melanoma cell lines. Mainly for the formation of microcells in melanoma cells. The drug-stimulated cells were then characterized in terms of their formation efficiency, morphology, and metabolic activity. Results: We observed the development of cancer microcells and green fluorescent protein (GFP) transfection efficiency after stress. In the time-lapse experiment, we observed microcell formation through a renewal process and GFP expression in the microcells. Additionally, the microcells were viable after anticancer treatment, as indicated by the nicotinamide adenine dinucleotide hydrogen phosphate (NADPH) enzyme activity assay results. Taken together, these findings indicate that cancer microcells are viable and capable of resisting the stress induced by anticancer drugs, and these cells are prone to chemical substance uptake from the environment. Conclusion: Microcells are not only common to a specific cancer type, but can be found in any tumor type. This study could help to understand cancer emergence and recurrence. The appearance of microcells in the studied cancer cell population could be an indicator of the individual anticancer therapy effectiveness and patient survival.Item Cultivation of 3D dermal tissue by application of autologous matrix(2020-02-01) Jakobsons, Ēriks; Ērglis, Kristaps; Ramata-Stunda, Anna; Blāķe, Ilze; Patetko, Liene; Tīcmane, Simona; Rupaine, Beatrise Lūcija; Ērglis, Mārtiņš; Ērgle, Māra; Strīķe, Eva; Strazdiņš, Uldis; Rubīns, Silvestrs; Rubīns, Andris; Ērglis, AndrejsThe most common reasons for major skin loss are thermal trauma — burns and scalds that can result in rapid, extensive, deep wounds as well as chronic non-healing wounds. Treatment using common techniques is poor and depending on the trauma level can result in death. There is a substantial need for skin integrity restoration. The main goal of this study was to develop an autologous 3D skin model that could eventually be translated into clinical applications. The study examined a variety of factors — extracellular matrix components, cell count, culture medium modification and role of structurally and functionally high-quality 3D skin dermis layer tissue culture production. The results of this study are an essential prerequisite to standardise the use of both clinical, as well as in vitro test systems. Dermal cell lines applied in the study were isolated form patient biopsies obtained at Pauls Stradiòð Clinical University Hospital. Blood plasma type AB was used for fibrin matrix formation. As catalysts, CaCl2 or calcium gluconate, and tranexamic acid were applied. 3D tissue functionality was assessed by evaluation of gene expression and changes in growth factor secretion. Fibrin matrix formulations with 1% and 1.5% CaCl2 and 5 mg, 7 mg and 10 mg tranexamic acid concentration were tested. Better matrix properties were observed with higher concentration of CaCl2 and tranexamic acid. Differences in levels of collagen gene expression and growth factor secretion were observed. Changes in levels of fibroblast growth factor and gene expression were observed in fibrin matrix samples and the surface-cultivated cell culture monolayer, but structural protein synthesis was not detected.Item In vitro impact preliminary assessment of airborne particulate from metalworking and woodworking industries(2021-10-12) Pavlovska, Ilona; Ramata-Stunda, Anna; Martinsone, Žanna; Boroduskis, Martins; Patetko, Liene; Mārtiņsone, Inese; Seile, Anita; Vanadziņš, Ivars; Institute of Occupational Safety and Environmental HealthInhalation is the main route of exposure to airborne pollutants. To evaluate the safety and assess the risks of occupational hazards different testing approaches are used. 3D airway epithelial tissues allow to mimic exposure conditions in vitro, generates human-relevant toxicology data, allows to elucidate the mode of action of pollutants. Gillian3500 pumps were used to collect the airborne particulate from woodworking and metalworking environments. EpiAirway tissues were used to model half working day (4 h), full working day (8 h), and 3 working day exposures to occupational pollutants. Tissue viability was assessed using an MTT assay. For preliminary assessment, RT-qPCR analyses were performed to analyze the expression of gelsolin, caspase-3, and IL-6. Tissue morphology was assessed by hematoxylin/eosin staining. An effect on the proliferation of lung epithelial cell line A549 was assessed. Acute exposure to workspace pollutants slightly affected tissue viability and did not change the morphology. No inhibiting effect was observed on the proliferation of A549 cells. Preliminary analysis showed that both types of particles suppressed the expression of gelsolin, with the effect of metalworking samples being more pronounced. A slight reduction in caspase-3 expression was observed. Particles from metalworking suppressed IL-6 expression.Item In Vitro Safety and Efficacy Evaluation of a Juniperus communis Callus Culture Extract and Matricaria recutita Processing Waste Extract Combination as a Cosmetic Ingredient(2024-01-18) Ramata-Stunda, Anna; Boroduskis, Martins; Pastare, Laura; Berga, Marta; Kienkas, Liene; Patetko, Liene; Skudrins, Gundars; Reihmane, Dace; Nakurte, IlvaFor skin health promotion and cosmetic applications, combinations of plant cell extracts are extensively utilized. As most natural ingredient suppliers offer crude extracts from individual plants or specific isolated compounds, the potential interactions between them are assessed in the development phase of cosmetic products. The industry seeks extract combinations that have undergone optimization and scrutiny for their bioactivities. This study presents a combination of two sustainably produced botanical ingredients and outlines their chemical composition, in vitro safety, and bioactivity for skin health enhancement. The amalgamation comprises the extract of Matricaria recutita processing waste and the extract from Juniperus communis callus culture. Chemical analysis revealed distinct compounds within the extracts, and their combination led to a broader array of potentially synergistic compounds. In vitro assessments on skin cells demonstrated that the combination possesses robust antioxidant properties and the ability to stimulate keratinocyte proliferation, along with regulating collagen type I and matrix metalloproteinase 1 (MMP-1) production by dermal fibroblasts. The identified traits of this combination render it an appealing cosmetic component. To the best of our knowledge, this represents the first case when the extracts derived from medicinal plant processing waste and biotechnological plant cell cultivation processes have been combined and evaluated for their bioactivity.Item New Modalities in Knee Osteoarthritis Treatment Using Autologous Bone Marrow-Derived Mononuclear Cells(2018-08-01) Gončars, Valdis; Kalnbērzs, Konstantīns; Jākobsons, Ērks; Briede, Ieva; Blums, Kristaps; Ērglis, Kristaps; Ērglis, Mārtiņš; Patetko, Liene; Muižnieks, Indriķis; Ērglis, AndrejsThe clinical effects on knee osteoarthritis (OA) symptoms and tissue structure were evaluated after bone marrow-derived mononuclear cell intraarticular injection. A group of 32 patients with 34 knee joints in stage II-III osteoarthritis were treated by intraarticular injection of mononuclear cell suspension. Clinical results were obtained by KOOS (Knee Osteoarthritis Outcome Score) and KSS (Knee Society Score) scores during a 12 months follow-up period. Radiological evaluation was performed using magnetic resonance imaging. A comparison with a control group of 28 patients treated with routinely used three hyaluronic acid intra-articular injections was made. No adverse effects were observed after the bone marrow derived mononuclear cells (BM-MNC) injection. At the end point of the follow up all score results had improved, compared to those at to the starting point. 65% of patients maintained minimal perceptible clinical improvement of the score results. The Whole Organ Magnetic Resonance Imaging Score showed improvement from 44.31 to 42.93 points (p < 0.05) during a 6-7 month period. Comparing score results to the control group, a statistically significant (p < 0.05) improvement in the KOOS pain subscale score at the 6 and 12 months was observed in the mononuclear cell group. BM-MNC injection leads to a decrease of knee OA symptoms and slows changes in structure of the degenerative joint tissue.