Ispitivanje dejstva niskotemperaturne atmosferske plazme in vitro na bakterije i mezenhimalne matične ćelije

Abstract

Poslednjih godina se intenzivno istražuje mogućnost primene različitih izvora niskotemperaturne atmosferske plazme za in vivo dezinfekciju i antimikrobnu terapiju u oblasti medicine i stomatologije. U ovom istraživanju kao izvor niskotemperaturne atmosferske plazme korišćena je modifikovana plazma igla razvijena u našoj zemlji i izvršeno je definisanje uslova pod kojima plazma stvorena ovim izvorom ispoljava baktericidni efekat, ali uz istovremeno očuvanje zdravog tkiva. Pokazano je da niskotemperaturna atmosferska plazma stvorena plazma iglom deluje snažno baktericidno na bakterije u suspenzijama različitih koncentracija, pri čemu je ovaj efekat u direktnoj zavisnosti od snage plazme, vremena izlaganja i početne koncentracije ispitivanih suspenzija bakterija. Utvrđeno je da su na dejstvo plazme bile osetljivije ispitivane Gram-negativne bakterije, Pseudomonas aeruginosa i Escherichia coli od Gram pozitivnih, Enterococcus faecalis i Staphylococcus aureus. Ispitivanjem uticaja niskotemperaturne atmosferske plazme na biofilm bakterija meticilin rezistentni Staphylococcus aureus i Streptococcus mutans pokazano je da plazma deluje inhibitorno na rast bakterija u biofilmu, dok ne utiče značajno na već formiran biofilm. Citotoksičnost niskotemperaturne atmosferske plazme ispitivana je na kulturi humanih mezenhimalnih matičnih ćelija poreklom iz periferne krvi. Utvrđeno je da primenjena pod uslovima pod kojim ispoljava snažno antimikrobno dejstvo, plazma ne utiče značajno na vijabilnost ovih ćelija niti dovodi do njihovog odlepljivanja od podloge. Prema podacima koji su dostupni u literaturi, do sada nije ispitivan uticaj niskotemperaturne atmosferske plazme na parodontalno tkivo. Stoga je u ovom radu po prvi put ispitivan uticaj niskotemperaturne atmosferske plazme na različite ćelijske funkcije humanih mezenhimalnih matičnih ćelija iz periodoncijuma. Humane mezenhimalne matične ćelije su uspešno izolovane i uspostavljena je njihova dugotrajna kultivacija. Ove ćelije su pokazale tipične karakteristike mezenhimalnih matičnih ćelija. Za ispitivanje uticaja niskotemperaturne atmosferske plazme na ćelijske funkcije humanih mezenhimalnih matičnih ćelija iz periodoncijuma odabrani su eksperimentalni uslovi pod kojima plazma ispoljava snažno antibakterijsko dejstvo na suspenzije bakterija i biofilm...

In recent years, there has been intensive research about the possibility of the application of low-temperature atmospheric plasma for in vivo desinfection and antimicrobial therapy in medicine and stomatology. In this study, as a source of non-thermal atmospheric plasma, a plasma needle device developed in our country was used and an investigation was carried out to define the conditions under which plasma generated by this source manifests bactericidal effect, while simultaneously preserving the healthy tissue. It was demonstrated that low-temperature atmospheric plasma generated by a plasma needle had a strong bactericidal effect on bacteria in suspensions of various concentrations, this effect being directly dependant on the plasma power, time of exposure and initial concentration of the examined suspensions. It was found that the examined Gram-negative bacteria, Pseudomonas aeruginosa and Escherichia coli, were more sensitive to plasma influence than the Gram-positive, Enterococcus faecalis and Staphylococcus aureus. By examining the effect of low-temperature atmospheric plasma on the bacteria biofilm of methicillin-resistant Staphylococcus aureus and Streptococcus mutans, it was demonstrated that it had an inhibitory effect on biofilm formation, however without significantly affecting the already formed biofilm. Cytotoxicity of lowtemperature atmospheric plasma was examined on the culture of human mesenchymal stem cells from peripheral blood. It was determined that, applied under the conditions under which it manifests strong antimicrobial effects, plasma did not have significant influence on the viability of these cells, nor caused detachment from the substrate. According to available literature data, the possibility of low-temperature atmospheric plasma application in parodontal therapy has not yet been examined. Therefore in this paper, the effect of non-thermal atmospheric plasma on different cell functions of mesenchymal stem cells isolated from human periodontal ligament was examined for the first time. Human mesenchymal stem cells were successfully isolated and their long-term cultivation was established. These cells showed typical human mesenchymal stem cell characteristics. To evaluate the influence of non-thermal atmospheric plasma on different cell functions of human periodontal-ligament derived mesenchymal stem cells, experimental conditions under which plasma expresses strong antibacterial effect on the bacteria suspensions and biofilm were applied...