Karakterizacija ljudskih osteoblasta izoliranih iz svježeg i smrznutog koštanog tkiva

master's thesis

• Title (english): Characterization of human osteoblasts isolated from fresh and frozen bone tissue
• Author: Zlata Sošić
• Mentor: Marija Zekušić (mentor)
• Mentor: Inga Urlić (komentor)
• Committee member: Inga Urlić (predsjednik povjerenstva)
• Committee member: Petar Kružić (član povjerenstva)
• Committee member: Sandra Hudina (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2023-08-31, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Poznavanje građe koštanog tkiva i razumijevanje funkcije stanica i signalnih molekula važno
je za napredak regenerativne medicine. Kosti su najčešće transplantirana muskuloskeletna
tkiva. Autologne kosti smatraju se zlatnim standardnom osiguravajući osteogenezu,
osteokondukciju i osteoindukciju. Krioprezervacijom 10%-tnim dimetil sulfoksidom (DMSO),
tkiva se dugotrajno pohranjuju na ultraniskim temperaturama < -80 °C s ciljem održavanja
vitalnosti stanica (npr. autologni osteohondralni graft). Korištenje autolognih kostiju je
limitirano, zato se koristi alogeno koštano tkivo (npr. glava bedrene kosti) osiguravajući
osteokondukciju i osteoindukciju. Pohranjuju se bez korištenja krioprotektanta na ultraniskim
temperaturama. Zamrzavanje ima mali utjecaj na mehanička svojstva koštanog tkiva te
umanjuje imunogenost. Kost je izgrađena od stanica osteoblasta, osteoklasta, osteocita i stanica
koje oblažu površinu kosti. Osteoblasti imaju ulogu u izgradnji, remodeliranju i regeneraciji
koštanog tkiva. Obzirom na potencijalno štetne i citotoksične učinke DMSO-a na pacijente,
važno je ispitati njegovo djelovanje na koštano tkivo i stanice jer uloga krioprotektanta kod
pohrane muskuloskeletnih tkiva nije u potpunosti jasna. U ovome istraživanju se ispituju i
uspoređuju citološka i molekularna svojstva osteoblasta izoliranih iz svježeg koštanog tkiva te
tkiva pohranjenog s i bez DMSO-a u periodu od 15 i 30 dana. Morfologija, vijabilnost,
mogućnost proliferacije i genska ekspresija osnovne su karakteristike stanica na koje
krioprotektant može imati utjecaja.

Abstract (english)

Knowing the structure of bone tissue and understanding the function of cells and signaling
molecules is important for the progress of regenerative medicine. Bones are the most frequently
transplanted musculoskeletal tissues. Autologous bone is considered the gold standard,
providing osteogenesis, osteoconduction and osteoinduction. By cryopreservation with 10%
dimethyl sulfoxide (DMSO), tissues are stored for a long time at ultra-low temperatures < -80
°C with the aim of maintaining cell vitality (eg autologous osteochondral graft). The use of
autologous bones is limited, that's why allogeneic bone tissue (eg femoral head) is used,
ensuring osteoconduction and osteoinduction. They are stored without the use of
cryoprotectants at ultra-low temperatures. Freezing has little effect on the mechanical properties
of bone tissue and reduces immunogenicity. Bone is made up of osteoblasts, osteoclasts,
osteocytes and bone lining cells. Osteoblasts play a role in the construction, remodeling and
regeneration of bone tissue. Considering the potentially harmful and cytotoxic effects of DMSO
on patients, it is important to examine its effect on bone tissue and cells, because the role of
cryoprotectants in the storage of musculoskeletal tissues is not entirely clear. In this research,
the cytological and molecular properties of osteoblasts isolated from fresh bone tissue and tissue
stored with and without DMSO for a period of 15 and 30 days are examined and compared.
Morphology, viability, the possibility of proliferation and gene expression are the basic
characteristics of cells on which the cryoprotectant can have an effect.

• Keywords: dimetil sulfoksid; koštani fragmenti; krioprezervacija; osteogena diferencijacija
• Keywords (english): dimethyl sulfoxide; bone fragments; cryopreservation; osteogenic differentiation
• Language: croatian
• URN:NBN: urn:nbn:hr:217:655950
• 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: Embargoed access; Embargo expiration date: 2025-08-31
• Created on: 2023-09-12 11:16:51