1. Razvoj novih metod izdelave in karakterizacije mikrožilja v in vitro modelihJernej Vajda, 2025, doctoral dissertation Abstract: Doktorska disertacija obravnava optimizacijo postopkov, materialov in analiznih metod v tkivnem inženirstvu mikrožilja z multidisciplinarnim pristopom, ki poleg uveljavljenih bioloških in kemijskih metod vključuje tudi računalniške simulacije, mehanske teste in nanotomografijo. V okviru raziskovalnega dela smo optimizirali linearnost ekstruzije mehanskih ekstruzijskih 3D tiskalnikov, s čimer smo dosegli boljše in bolj ponovljive rezultate 3D (bio)tiska vzorcev. Pri natisnjenih vzorcih smo primerjali uporabo CaCl2 in SrCl₂ kot ionskih zamreževalcev, pri čemer se je izkazalo, da je uporaba SrCl2 ob primerljivi celični metabolni aktivnosti izboljšala mehanske lastnosti materialov, kombinacija Sr²⁺ in Ca²⁺ pa bi lahko omogočala dodatno prilagoditev mehanskih lastnosti materialov. Uporaba Sr²⁺ hkrati zmanjša tudi vpliv kalcijevih ionov na signalizacijo celic, kar je ključno za simuliranje biološkega okolja. Na drugi strani smo s simulacijami računalniške dinamike tekočin določili mejne pogoje za razvoj peristaltične črpalke,
s katero smo eksperimentalno ovrednotili vpliv strižnih napetosti na endotelijske celice in validirali simulirane rezultate. Ločeno smo na endotelijskih celicah iz različnih virov in na podpornih celicah – kožnih fibroblastih – preiskovali lastnosti materialov in njihov vpliv na viabilnost, ohranitev celičnega fenotipa in funkcionalnosti ter celokupno metabolno aktivnost. Rezultati študij potrjujejo, da je z natančno optimizacijo procesov in materialov mogoče izboljšati funkcionalnost tkivnih in vitro modelov, kar odpira nove možnosti za napredek v tkivnem inženirstvu in regenerativni medicini.
Keywords: mikrožilje, in vitro model, 3D tisk, računalniška dinamika tekočin, nanotomografija
Published in DKUM: 04.07.2025; Views: 0; Downloads: 4
 Full text (52,49 MB) |
2. Novel small-molecule inhibitors of the SARS-CoV-2 spike protein binding to neuropilin 1Anja Kolarič, Marko Jukič, Urban Bren, 2022, original scientific article Abstract: Furin cleavage of the SARS-CoV-2 spike protein results in a polybasic terminal sequence
termed the C-end rule (CendR), which is responsible for the binding to neuropilin 1 (NRP1), enhancing
viral infectivity and entry into the cell. Here we report the identification of 20 small-molecule
inhibitors that emerged from a virtual screening of nearly 950,000 drug-like compounds that bind
with high probability to the CendR-binding pocket of NRP1. In a spike NRP1 binding assay, two of
these compounds displayed a stronger inhibition of spike protein binding to NRP1 than the known
NRP1 antagonist EG00229, for which the inhibition of the CendR peptide binding to NRP1 was
also experimentally confirmed. These compounds present a good starting point for the design of
small-molecule antagonists against the SARS-CoV-2 viral entry.
Keywords: neuropilin 1, SARS-CoV-2, COVID-19, spike binding inhibitors, virtual screening, small-molecule antagonists, molecular docking, in vitro binding assay
Published in DKUM: 09.05.2025; Views: 0; Downloads: 5
Full text (6,27 MB)
This document has many files! More... |
3. Copper sulfate elicitation effect on biomass production, phenolic compounds accumulation, and antioxidant activity of Morus nigra L. stem node cultureJan Senekovič, Špela Jelen, Andreja Urbanek Krajnc, 2025, original scientific article Abstract: Phenolic compounds are strong antioxidant and antibacterial agents with great pharmacological, medicinal, nutritional, and industrial value. The potential of Morus nigra in stem node culture was investigated for the production of phenolic compounds and their elicitation with CuSO4. Individual phenolic compounds in the samples were identified and quantified by using HPLC-PDA and HPLC-MS methods, while the content of total phenolic compounds, the content of total flavonoids, and the antioxidant activity of methanolic extracts were evaluated spectrophotometrically. The highest fresh and dry weights were obtained in plantlets treated with 0.5 mM CuSO4 for 42 days. The highest total phenolic content, total flavonoid content, and antioxidant activity of the extracts were determined in stem node cultures treated with 3 mM CuSO4 for 42 days. Under the latter conditions, the predominant representatives of the caffeoylquinic acids, p-coumaric acid derivatives, kaempferol derivatives, and quercetin derivatives also achieved the highest content. The most abundant phenolic compound in all samples was the chlorogenic acid. The nodal culture of M. nigra elicited with CuSO4 could potentially be used for the industrial production of phenolic compounds, especially caffeoylquinic acids. Moreover, considering the biochemical response to CuSO4 treatment and the ability to tolerate and accumulate copper, the potential application of M. nigra in phytoremediation is also highlighted.
Keywords: antioxidative potential, black mulberry, copper stress, elicitation of phenolics, flavonoids, in vitro culture, nodal culture, phenolic acids
Published in DKUM: 25.03.2025; Views: 0; Downloads: 10
 Link to file |
4. Comparison of conventional and supercritical isolation methods from bee pollen and their effects on the concentrations of bioactive compoundsTaja Žitek Makoter, Vesna Postružnik, Anastasia Kyriakoudi, Elli Pantazi, Natasa Kalogiouri, Ioannis Mourtzinos, Željko Knez, Maša Knez Marevci, 2025, original scientific article Abstract: The present work represents a profound investigation of material pretreatment, comparison of different isolation methods (conventional and supercritical), and characterization of phenolic compounds and amino acids from bee pollen. The determination of amino acids was carried out after developing a novel hydrophilic interaction chromatography combined with quadrupole time-of-flight tandem mass spectrometric method (HILIC-QTOF-MS/MS) with limits of detection over the range 1 mg/kg -5 mg/kg and recoveries over the range 87.6% - 97.1%. The optimal conditions for the isolation of amino acids are in the range of temperatures between 25°C to 55°C and at pressures between 300 bar and 350 bar. It can be concluded that the extraction temperature and the pressure affect antioxidant activity. The favored conditions are lower temperatures and higher pressures.
Keywords: bee pollen, in vitro antioxidant activity, HILIC-LC-QTOF-MS.MS, amino acids, WM-266-4 cells
Published in DKUM: 20.03.2025; Views: 0; Downloads: 2
Full text (1,27 MB) |
5. Design of in vitro hair follicles for different applications in the treatment of alopecia : a reviewMatej Žnidarič, Žan Michel Žurga, Uroš Maver, 2021, review article Abstract: The hair research field has seen great improvement in recent decades, with in vitro hair follicle (HF) models being extensively developed. However, due to the cellular complexity and number of various molecular interactions that must be coordinated, a fully functional in vitro model of HFs remains elusive. The most common bioengineering approach to grow HFs in vitro is to manipulate their features on cellular and molecular levels, with dermal papilla cells being the main focus. In this study, we focus on providing a better understanding of HFs in general and how they behave in vitro. The first part of the review presents skin morphology with an emphasis on HFs and hair loss. The remainder of the paper evaluates cells, materials, and methods of in vitro growth of HFs. Lastly, in vitro models and assays for evaluating the effects of active compounds on alopecia and hair growth are presented, with the final emphasis on applications of in vitro HFs in hair transplantation. Since the growth of in vitro HFs is a complicated procedure, there is still a great number of unanswered questions aimed at understanding the long-term cycling of HFs without losing inductivity. Incorporating other regions of HFs that lead to the successful formation of different hair classes remains a difficult challenge.
Keywords: hair follicle, alopecia, dermal papilla cells, hair transplantation, in vitro hair follicle models
Published in DKUM: 11.03.2025; Views: 0; Downloads: 3
Full text (2,39 MB)
This document has many files! More... |
6. An overview of the current state of cell viability assessment methods using OECD classificationEneko Madorran, Miha Ambrož, Jure Knez, Monika Sobočan, 2025, review article Abstract: Over the past century, numerous methods for assessing cell viability have been
developed, and there are many different ways to categorize these methods accordingly. We
have chosen to use the Organisation for Economic Co-operation and Development (OECD)
classification due to its regulatory importance. The OECD categorizes these methods
into four groups: non-invasive cell structure damage, invasive cell structure damage, cell
growth, and cellular metabolism. Despite the variety of cell viability methods available,
they can all be categorized within these four groups, except for two novel methods based
on the cell membrane potential, which we added to the list. Each method operates on
different principles and has its own advantages and disadvantages, making it essential
for researchers to choose the method that best fits their experimental design. This review
aims to assist researchers in making this decision by describing these methods regarding
their potential use and providing direct references to the cell viability assessment methods.
Additionally, we use the OECD classification to facilitate potential regulatory use and to
highlight the need for adding a new category to their list.
Keywords: cell viability, cell-based methods, in vitro toxicology, OECD cell viability classification
Published in DKUM: 13.02.2025; Views: 0; Downloads: 7
Full text (2,77 MB)
This document has many files! More... |
7. Microbiota in vitro modulated with polyphenols shows decreased colonization resistance against Clostridioides difficile but can neutralize cytotoxicityAleksander Mahnič, Jennifer Auchtung, Nataša Poklar Ulrih, Robert A. Britton, Maja Rupnik, 2020, original scientific article Abstract: While the knowledge on gut microbiota - C. difficile interactions has improved over the years, the understanding of the underlying mechanisms providing colonization resistance as well as preventative measures against the infection remain incomplete. In this study the antibiotic clindamycin and polyphenol extracts from pomegranate and blueberries were used individually and in combination to modulate fecal microbial communities in minibioreactor arrays (MBRA). Modulated communities were inoculated with C. difficile (ribotype 027). Subsequent 7-day periodical monitoring included evaluation of C. difficile growth and activity of toxins TcdA and TcdB as well as analysis of MBRA bacterial community structure (V3V4 16%S metagenomics). Polyphenols affected multiple commensal bacterial groups and showed different synergistic and antagonistic effects in combination with clindamycin. Exposure to either clindamycin or polyphenols led to the loss of colonization resistance against C. difficile. The successful growth of C. difficile was most significantly correlated with the decrease in Collinsella and Lachnospiraceae. Additionally, we demonstrated that Clostridium sporogenes decreased the activity of both C. difficile toxins TcdA and TcdB. The feature was shown to be common among distinct C. sporogenes strains and could potentially be applicable as a non-antibiotic agent for the alleviation of C. difficile infection.
Keywords: microbiota in vitro, Clostridioides difficile, resistance, cytotoxicity
Published in DKUM: 29.01.2025; Views: 0; Downloads: 5
Full text (1,67 MB)
This document has many files! More... |
8. Razvoj in vitro modela kožeMaja Sever, Dominik Škrinjar, 2024, final research report Abstract: Tridimenzionalni (3D) biotiskani modeli kože se vse bolj uveljavljajo v predkliničnih in kliničnih raziskavah kot alternativa testiranju na živalih. Ključno za uspeh teh raziskav je zagotavljanje ponovljivosti in primerljivosti rezultatov, kar zahteva razvoj zanesljivih hidrogelov, ki omogočajo stabilno vključitev celic v 3D biotisk. Eden največjih izzivov pri tem je zagotavljanje konsistentnosti lastnosti hidrogela, ki je temeljni material za izdelavo 3D tiskanega in vitro modela kože, potrebnega za različne faze raziskav. Namen te raziskave je bil preučiti možnost priprave večje serije hidrogela ter raziskati načine njihovega dolgoročnega shranjevanja, s poudarkom na tehnikah zamrzovanja, z namenom ohranitve njihovih fizikalno-kemijskih lastnosti. Tako bi omogočili uporabo enotne serije hidrogela skozi celoten razvoj in vitro modela kože, kar bi zmanjšalo variacije, ki nastajajo pri posameznih pripravah hidrogelov za različne raziskovalne metode. Osredotočili smo se na vpliv temperature (-20 °C in -80 °C) ter trajanja shranjevanja (7 in 29 dni) na ključne lastnosti hidrogelov, kot so viskoznost, poroznost, sposobnost nabrekanja, degradacija in mehanske lastnosti po odmrznitvi. Potrditev teh hipotez bi prispevala k večji ponovljivosti in natančnosti 3D biotiskljivih in vitro modelov kože.
Keywords: in vitro model kože, 3D biotiskanje, hidrogel, predkinična in klinična raziskava
Published in DKUM: 24.01.2025; Views: 0; Downloads: 11
Full text (2,90 MB) |
9. Kratek pregled platform za ADME testiranjeTina Maver, Boštjan Vihar, Uroš Maver, 2024, original scientific article Abstract: V zadnjem času je bil dosežen pomemben napredek pri razvoju ADME (absorpcija, distribucija, metabolizem, ekskrecija) modelov, vendar izziv ostaja vzpostaviti platforme, ki bi zmanjšale testiranje na živalih in stroške raziskav. Naraščajoča pomembnost farmakokinetičnih interakcij poudarja potrebo po zanesljivih in
ponovljivih ADME modelih, ki so vse bolj ključni za razvoj zdravil in zagotavljanje varnosti z željo po preprečevanju
resnih kliničnih zapletov in hospitalizacije.
Keywords: ADME, drug interaction studies, in vitro models, pharmacokinetics, multi-organ models
Published in DKUM: 07.01.2025; Views: 0; Downloads: 23
Full text (405,16 KB)
This document has many files! More...
This document is also a collection of 1 document! |
10. Pravna ureditev oploditve z biomedicinsko pomočjo v luči raziskav na človeških zarodkih in njihovih matičnih celicah : magistrsko deloLara Krneža, 2024, master's thesis Abstract: V 20. stoletju so z namenom razvoja medicine, preprečevanja genetsko bolnih potomcev in izboljšanja vrste izvajali neetične poskuse na ljudeh. Ti so bili opravljeni brez privolitve žrtev. Zaradi vedno večje ozaveščenosti družbe o človekovih pravicah in napredka pravne znanosti, se je po drugi svetovni vojni začela sprejemati zakonodaja, ki je postavila etične smernice za izvajanje raziskav na ljudeh. Leta 1947 je bil sprejet Nürnberški kodeks, leto kasneje pa je Svetovno zdravniško združenje sprejelo Ženevsko deklaracijo o zdravniških dolžnostih. Pomembne svetovne organizacije so zatem podpisale še Evropsko konvencijo o varstvu človekovih pravic in temeljnih svoboščin, Helsinško deklaracijo o etičnih načelih medicinskih raziskav na ljudeh, Oviedsko konvencijo, Splošno deklaracijo o človeškem genomu in človekovih pravicah, Splošno deklaracijo o bioetiki in človekovih pravicah in Listino EU o temeljnih pravicah. Vsi navedeni pravni akti so pomembno vplivali na pravno ureditev raziskav (na človeških zarodkih in njihovih matičnih celicah) v Sloveniji.
Opravljanje medicinskih poskusov na ljudeh brez njihove svobodne privolitve je pri nas prepovedano že z Ustavo Republike Slovenije. Ta določa tudi ostale ustavne kategorije, v luči katerih se morajo izvajati raziskave. Biomedicinske raziskave in kazniva dejanja v zvezi z njimi so opredeljena v Kazenskem zakoniku, ki prepoveduje kloniranje, ustvarjanje človeških zarodkov v raziskovalne (in druge) namene ter izvajanje drugih biomedicinskih posegov. Pravice pacientov ureja Zakon o pacientovih pravicah. Za magistrsko nalogo sta ključnega pomena še Zakon o zdravljenju neplodnosti in postopkih oploditve z biomedicinsko pomočjo, ki ureja raziskave na zarodkih »in vitro«, pridobivanje in raziskovanje na embrionalnih matičnih celicah in Kodeks zdravniške etike, ki določa etične standarde za zdravnike pri izvajanju raziskav na pacientih.
Pravna teorija in sodna praksa se pri pravnem urejanju raziskav na človeških zarodkih in njihovih matičnih celicah pogosto srečujeta s kolizijo med dvema ustavnopravnima kategorijama, pravico do življenja (zarodka) in svobodo znanosti. Do te kolizije pogosto prihaja zaradi nerešenega vprašanja o pravnem statusu človeškega zarodka. Evropsko sodišče za človekove pravice je o tem odločalo že večkrat, a enotne rešitve na to vprašanje doslej še ni podalo. Pomembni precedensi ESČP v zvezi z raziskavami na človeških zarodkih in pravnim statusom zarodka so zadeve Evans proti Združenemu Kraljestvu, Vo proti Franciji in Parillo proti Italiji.
Keywords: medicinsko pravo, človekove pravice, medicinska etika, medicinske raziskave, poskusi na ljudeh, evgenika, privolitev, znanstvene raziskave na in vitro zarodkih, in vitro oploditev
Published in DKUM: 11.12.2024; Views: 0; Downloads: 129
Full text (877,37 KB) |
