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Vpliv zunajtelesnega krvnega obtoka samega in v kombinaciji z zunajtelesno imunoadsorpcijo oziroma metilprednizolonom na zunajcelične vezikle pri srčnih operacijah
Jevnikar, Elizabeta (Avtor), Černe, Katarina (Mentor) Več o mentorju... Povezava se odpre v novem oknu, Podbregar, Matej (Komentor)

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Izvleček
Zunajtelesni krvni obtok (ZTKO) v obliki srčno-pljučnega obvoda je ključen za večino operacij na odprtem srcu. Z njim je povezan sistemski vnetni odgovor, ki lahko vodi do pooperativnih zapletov ali smrti. Potencialni biološki označevalci, ki bi lahko predvideli z ZTKO povezane zaplete, so zunajcelični vezikli (ZV) – heterogena skupina z membrano obdanih struktur, ki se sproščajo iz celic. Obstaja potreba po podrobnejšem razumevanju povezave med ZTKO in ZV in vpliva dodatnih ukrepov, kot sta na primer uporaba zunajtelesnega adsorberja citokinov CytoSorb® (CytoSorb®, CytoSorbents Inc., ZDA) ter administracija kortikosteroidov. Glavni namen magistrskega dela je bil proučiti vpliv samega ZTKO, ZTKO z zunajtelesno imunoadsorpcijo in ZTKO z administracijo metilprednizolona na velikost ter raven ZV v krvi pacientov, ki so prestali kompleksno srčno operacijo. Proučili smo tudi morebitno korelacijo rezultatov s kliničnimi podatki. Sekundarni cilj je bil optimizacija postopkov za karakterizacijo in kvantifikacijo ZV z analizo sledenja nanodelcem (NTA) ter pretočno citometrijo. V analizo smo vključili vzorce krvi 23 bolnikov, ki so bili operirani na srcu na SPS Kirurški kliniki, UKCLJ. Bolniki so bili razdeljeni v kontrolno skupino (9 bolnikov brez kapsule CytoSorb® in brez kortikosteroidov), skupino s kapsulo (8 bolnikov, pri katerih je bila med ZTKO uporabljena kapsula CytoSorb®) in kortikosteroidno skupino (6 bolnikov, ki so med ZTKO prejeli metilprednizolon). Vzorci krvi so bili odvzeti pred indukcijo v anestezijo (pred ZTKO) in po ZTKO (po ZTKO). Po pripravi vzorcev za analizo je sledila analiza ZV z NTA in pretočno citometrijo. Začetne velikosti in ravni ZV v vzorcih pred ZTKO so se med bolniki zelo razlikovale. Velikost ZV se je po operaciji ZTKO statistično značilno povišala (p < 0,001). Koncentracija ZV se po ZTKO ni statistično značilno spremenila (p = 0,298). Čeprav se je koncentracija celokupnih bioloških nanodelcev po ZTKO statistično značilno zmanjšala (p < 0,001), to zmanjšanje ne odraža sprememb v ravni ZV. Po ZTKO se je statistično značilno zmanjšala tudi fluorescenca ZV (p = 0,011), kar kaže na zmanjšanje esterazne aktivnosti znotraj ZV. Trend spremembe ravni ZV zaradi ZTKO ni bil enak pri vseh bolnikih, medtem ko je bilo pri vseh bolnikih prisotno povečanje velikosti ZV po ZTKO. Zunajtelesna imunoadsorpcija ni dodatno vplivala na povečanje velikosti ZV zaradi ZTKO (p < 0,001; kontrola p < 0,001), je pa dodatno vplivala na povečanje koncentracije ZV (p = 0,016; kontrola p = 0,466). Zunajtelesna imunoadsorcpija je dodatno vplivala na znižanje fluorescence (esterazne aktivnosti) ZV, ki je bilo statistično značilno le v skupini s kapsulo (p < 0,001). Administracija metilprednizolona ni dodatno vplivala na povečanje velikosti ZV, povzročeno z ZTKO (p < 0,001, kontrola p < 0,001). Metilprednizolon ni dodatno vplival na spremembo v koncentraciji ZV zaradi ZTKO (p = 0,566, kontrola p = 0,466). Administracija metilprednizolona ni dodatno vplivala na fluorescenco ZV oz. esterazno aktivnost (p = 0,059, kontrola p = 0,507). Velikost in raven ZV ter njihova fluorescenca v vzorcih krvi bolnikov pred ZTKO in po njem so bili povezani s kliničnimi podatki bolnikov. Prav tako so s kliničnimi podatki korelirale relativne spremembe v parametrih, ki karakterizirajo ZV. Dokazali smo, da operacija z ZTKO povzroči spremembe v ZV, dodatno pa lahko vplivajo tudi dodatni ukrepi, kot je zunajtelesna imunoadsorpcija. Dodatnega učinka na ZV zaradi administracije metilprednizolona nismo dokazali. Potrebne so nadaljnje raziskave, idealno na večjem številu bolnikov, ki bi podrobneje proučile vpliv ZTKO in dodatnih ukrepov na ZV. Pokazali smo tudi, da so ZV povezani s kliničnimi podatki, kar odpira možnosti njihove uporabe kot bioloških označevalcev.

Jezik:Slovenski jezik
Ključne besede:zunajtelesni krvni obtok, zunajcelični vezikli, zunajtelesna imunoadsorpcija, Cytosorb, metilprednizolon
Vrsta gradiva:Magistrsko delo/naloga (mb22)
Tipologija:2.09 - Magistrsko delo
Organizacija:FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Leto izida:2020
COBISS.SI-ID:36540931 Povezava se odpre v novem oknu
Število ogledov:146
Število prenosov:64
Metapodatki:XML RDF-CHPDL DC-XML DC-RDF
 
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Sekundarni jezik

Jezik:Angleški jezik
Naslov:The impact of extracorporeal blood circulation alone and in combination with extracorporeal immunoadsorption or methylprednisolone on extracellular vesicles in cardiac surgery
Izvleček:
Extracorporeal blood circulation (EBC) in the form of cardiopulmonary bypass is essential for most open-heart surgeries. EBC often triggers a systemic inflammatory response, which can lead to postoperative complications or death. Extracellular vesicles (EVs), heterogeneous populations of membrane vesicles released by the cells, are potential biomarkers of adverse events during EBC. There is a need for a more detailed understanding of the relation between EBC and EVs and the effect of additional measures such as extracorporeal cytokine adsorber CytoSorb® (CytoSorb®, CytoSorbents Inc., USA) and corticosteroid administration. The main purpose of the master’s thesis was to study the impact of extracorporeal blood circulation alone and in combination with extracorporeal immunoadsorption or methylprednisolone on EVs levels and their size in blood samples of patients who have undergone a complex heart surgery. We also examined potential correlation of the results with clinical data. A secondary goal was to optimize the protocols for EVs characterization and quantification with the nanoparticle tracking analysis (NTA) and flow cytometry. Blood samples of 23 patients who have undergone heart surgery on SPS Kirurška klinika, University Medical Centre Ljubljana, were included in the research. Patients were divided in the control group (9 patients, without CytoSorb® capsule and without corticosteroids during EBC), Cytosorb group (8 patients, a capsule CytoSorb® was used during EBC), and the corticosteroid group (6 patients receiving methylprednisolone during EBC). The blood samples were taken before the induction into anaesthesia (pre-EBC) and after EBC (post-EBC). After the preparation of the samples, the analysis with NTA and flow cytometry followed. Preoperative EV levels and their size in the pre-EBC samples varied greatly between patients. EV size increased significantly after the surgery with EBC (p < 0,001). EV concentration did not change significantly after EBC (p = 0,298). Although the total biological nanoparticle concentration decreased statistically significantly after the EBC (p <0,001), this decrease does not reflect changes in the level of EV. EV fluorescence decreased statistically significantly after the EBC (p = 0,011) which indicates a decrease in esterase activity within the EVs. The trend of change in the EV levels due to EBC was not the same in all patients, while there was an increase in the size of the EVs after EBC in all patients. Extracorporeal immunoadsorption did not additionally affect the increase in EV size due to EBC (p < 0,001; control p < 0,001); however, it did additionally affect the increase of EV concentration (p = 0,016; control p = 0,466). Extracorporeal immunoadsorption had an additional effect on the decrease in fluorescence (esterase activity) of EVs, which was statistically significant only in the Cytosorb group (p < 0,001). Administration of methylprednisolone did not additionally affect the increase in EV size caused by EBC (p < 0,001; control p < 0,001). Methylprednisolone had no additional effect on the change in EV concentration due to EBC (p = 0,566, control p = 0,466). Administration of methylprednisolone did not further affect EV fluorescence or esterase activity (p = 0,059, control p = 0,507). The EV levels, their size and fluorescence in the pre-EBC and post-EBC blood samples correlated with the clinical data of patients. Relative changes in parameters that characterize EV correlated with clinical data, as well. We have demonstrated that surgery with EBC causes changes in EVs, and supplementary measures such as extracorporeal immunoadsorption may have an additional effect. No additional effect on the EVs due to methylprednisolone administration was demonstrated. Further research is needed, ideally on a larger number of patients, to examine the impact of EBC and additional measures on EVs in more detail. We have also showed that EVs are associated with clinical data, which opens up the possibility of their use as biological markers.

Ključne besede:extracorporeal blood circulation, extracellular vesicles, extracorporeal immunoadsorption, Cytosorb, methylprednisolone

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