Modeliranje utjecaja ekstrakta ličinke Hermetia illucens na cijeljenje rana in vitro

master's thesis

• Title (english): In vitro modelling of the infuence of extracts from Hermetia illucens larvae on wound healing
• Author: Mislav Majnarić
• Mentor: Herwig O. Gutzeit (mentor)
• Mentor: Aleksandar Vojta (mentor)
• Committee member: Aleksandar Vojta (predsjednik povjerenstva)
• Committee member: Damjan Franjević (član povjerenstva)
• Committee member: Martina Šeruga Musić (član povjerenstva)
• Committee member: Vlatka Zoldoš (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2016-09-21, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Ličinke crne vojničke muhe (Black soldier fly), Hermetia illucens, su strvinari koji žive u izrazito neprijatnom okolišu, poput gnojiva i komposta, naseljenima bakterijama i gljivicama. Strvine uginulih životinja i trule biljke mogu se razgraditi ličinkama crne vojničke muhe. Ovakve biološke karakteristike upućuju na to da su ličinke vrste Hermetia illucens bogate antimikrobnim peptidima (AMP) i ostalim tvarima koje posjeduju aktivnost za borbu protiv rezistentnih bakterijskih sojeva. Cijeljenje rana je složen proces koji se sastoji od četiri preklapajuće ali strogo definirane faze: hemostaze, upalne faze, faze cijeljenja (proliferacije) i faze remodelacije (maturacije i epitelizacije). Sve faze procesa cijeljenja rana ovisne su o lučenju različitih staničnih spojeva poput: faktora rasta, kemokina, citokina, proteinaza, proteina izvanstaničnog matriksa. In vitro proces cijeljenja rana može se pratiti uporabom ovih različitih staničnih spojeva kao molekularnih markera. Fokus ovog istraživanja je obogaćivanje ekstrakata ličinki vrste Hermetia illucens u svrhu dobivanja pročišćenih AMP-ova i ostalih antibiotskih spojeva te in vitro praćenje utjecaja ekstrakata iz ličinki vrste H. illucens na rast bakterija te na citotoksičnost ljudskih stanica. Ekstrakti su pokazali visok kapacitet inhibicije bakterijskog rasta, posebice vrste Pseudomonas fluorescens. Štoviše, većina esktrakata koji su inhibirali bakterijski rast nisu se pokazali citotoksičnim za ljudske stanice.

Abstract (english)

Larvae of black soldier fly, Hermetia illucens, are scavengers who live in extremely unpleasant and harsh environments, such as manure and compost, populated by bacteria and fungi. Carcasses of dead animals and rotting plants could be degraded by larvae of black soldier fly. These biological characteristics suggest that the larvae of black soldier fly are rich in antimicrobial peptides (AMPs) and other substances which possess the activity to fight against resistant bacterial strains.
Wound healing is a complex process consisting of four overlapping but strictly defined phases: hemostasis, inflammatory phase, proliferative phase and the remodelling phase (maturation and epithelialization). All phases of wound healing are dependent on the secretion of a variety of cellular compounds such as growth factors, chemokines, cytokines, proteinases, and extracellular matrix proteins. In vitro process of wound healing could be monitored using these different cellular compounds as molecular markers. Focus of this research is enrichment of extracts from H. illucens larvae in order to obtain purified AMPs and other antibiotic compounds and in vitro monitoring of the impact of extracts from H. illucens larvae on bacterial growth and cytotoxicity of human cells. Extracts showed high capacity of inhibition of bacterial growth, especially species Pseudomonas fluorescens. Moreover, majority of extracts which inhibited bacterial growth did not show citotoxic effect on human cells.

• Keywords: Crna vojnička muha (Hermetia illucens); ekstrakti ličinke; HepG2- ljudska stanična linija raka jetre; normalni ljudski fibroblasti kože; antimikrobini efekt; cijeljenje rana
• Keywords (english): Black soldier fly (Hermetia illucens); larval extracts; Hep G2-human liver cancer cell line; Normal human dermal fibroblasts (NHDF); antimicrobial effect; wound healing
• Language: croatian
• URN:NBN: urn:nbn:hr:217:250794
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-05-08 09:46:07