Novel ex vivo human osteochondral explant model of knee and spine osteoarthritis enables assessment of inflammatory and drug treatment responses

master's thesis

• Title (croatian): Novi ex vivo humani osteohondralni eksplantacijski model koljena i kralježnice omogućuju procjenu upalnog i terapijskog odgovora
• Author: Doria Jurić
• Mentor: Mislav Jelić (mentor)
• Committee member: Mislav Jelić (predsjednik povjerenstva)
• Committee member: Ivan Bojanić (član povjerenstva)
• Committee member: Tomislav Đapić (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; (Department of Orthopedics); Zagreb
• Defense date and country: 2019-07-12, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences; Orthopedics

Abstract

Osteoarthritis of the knee and spine is highly prevalent in the modern society, yet a disease-modifying pharmacological treatment remains an unmet clinical need. A major challenge for drug development includes selection of appropriate preclinical models that accurately reflect clinical phenotypes of human disease. The aim of this study was to establish an ex vivo explant model of human knee and spine osteoarthritis that enables assessment of osteochondral tissue responses to inflammation and drug treatment. Equal-sized osteochondral fragments from knee and facet joints (both n = 6) were subjected to explant culture for 7 days in the presence of a toll-like receptor 4 (TLR4) agonist and an inhibitor of transforming growth factor-beta (TGF-β) receptor type I signaling. Markers of inflammation, interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), but not bone metabolism (pro-collagen-I) were significantly increased by treatment with TLR4 agonist. Targeting of TGF-β signaling resulted in a strong reduction of pro-collagen-I and significantly decreased IL-6 levels. MCP-1 secretion was increased, revealing a regulatory feedback mechanism between TGF-β and MCP-1 in joint tissues. These findings demonstrate proof-of-concept and feasibility of explant culture of human osteochondral specimens as a preclinical disease model, which might aid in definition and validation of diseasemodifying drug targets.

Abstract (croatian)

Osteoartritis koljena i kralježnice vrlo je čest u modernom društvu, no farmakološko liječenje koje utječe na bolest ostaje i dalje neispunjena klinička potreba. Glavni izazov za razvoj lijeka uključuje odabir odgovarajućih predkliničkih modela koji točno odražavaju kliničke fenotipove bolesti. Cilj ovog istraživanja bilo je utvrditi ex vivo model eksplantacije osteoartritisa ljudskog koljena i kralježnice koji omogućuju procjenu odgovora osteokondralnog tkiva na upalu i liječenje lijekovima. Osteokondralni fragmenti jednakih veličina iz zglobova koljena i faceta (oba n=6) podvrgnuti su kulturi eksplantacije 7 dana u prisutnosti agonista TLR4 i inhibitoraTGF-β receptora tipa I. Markeri upale, interleukin 6 (IL-6), monocitnikemoatraktantni
protein-1, ali ne i metabolizam kosti (pro-kolagen-1) bili su značajno povišeni liječenjem s TLR4 agonistom. Ciljanje TGF-β signalizacije rezultiralo je jakom redukcijom pro-kolagena-1 i značajno smanjilo razine IL-6. MCP-1 sekrecija je povećana, otkrivajući regulatorni mehanizam povratne sprege između TGF-β i MCP-1 u zglobnom tkivu. Ovi nalazi dokazuju, koncept i izvedivost kulture eksplantacije ljudskih osteokondralnihuzoraka kao predklinički model bolesti, koji bi mogao pomoći u definiranju i validaciji ciljeva lijekova koji modificiraju bolesti.

• Keywords: osteoarthritis; osteochondral; experimental model; inflammation; bone metabolism; knee; spine
• Keywords (croatian): osteoartritis; osteokondralni; eksperimentalni model; upala; metabolizam kosti; koljeno; kralježnica
• Language: english
• URN:NBN: urn:nbn:hr:105:587056
• Study programme: Title: Medicine (in English language); Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica medicine (doktor/doktorica medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-07-13 08:47:02