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Research project (§ 26 & § 27)
Duration : 2017-03-01 - 2020-02-29

The proposed project is designed to test the hypotheses that successful cartilage regeneration requires the recapitulation of embryogenic processes and that the secretome of fetal cells can initiate fetal-like regeneration of cartilage. Osteoarthritis (OA), a degenerative joint disease characterized by progressive articular cartilage degeneration, is one of the leading causes of disability worldwide and is associated with a tremendous individual and socioeconomic burden. Adult articular cartilage (AAC) has limited intrinsic repair capacity and current medical treatment options provide only symptomatic relief without significantly altering the disease progression or restoring cartilage integrity. Therefore, injured cartilage does not regenerate but forms fibrocartilaginous repair tissue with impaired biomechanical properties which does not adequately substitute for hyaline cartilage, thereby precipitating the continuation of joint inflammation leading to chronic osteoarthritis. In contrast to AAC, fetal articular cartilage (FAC) subjected to partial thickness lesions fully regenerates. In addition, fetal cells transplanted into an adult organism have been shown to retain their regenerative potential in skin, liver, tendon and cartilage models. Information on regenerative healing in fetal animals may allow improvement of the healing response in mature tissue. The cell secretome has a key role in tissue regeneration but is poorly understood. An improved knowledge of the factors involved in healing is a key prerequisite for utilizing this potentially powerful tool for therapeutic applications. We intend to develop a novel, biomimetic treatment strategy, recapitulating aspects of fetal articular cartilage morphogenesis to achieve fetal-like regeneration of adult articular cartilage. We propose to carry out a comprehensive study comparing 1) fetal (gestational day 80, term ~ 145 days of gestation) and adult cartilage healing in vivo 2) the effect of fetal chondrocytes, fetal mesenchymal stem cells (fMSCs), adult chondrocytes and adult bone marrow derived mesenchymal stem cells (aMSCs) and the secretome of these 4 cell types on adult articular chondrocyte proliferation, chondrogenic matrix production, and gene expression in vitro and 3) to identify key factors responsible for the induction of fetal healing rather than adult repair. The achieved knowledge will lead to further developmental work following up the proposed project, in order to establish an economically exploitable therapy approach, which will induce fetal healing in adult cartilage. The novel therapy will give the company partner involved in this project a competitive edge in this field at an international level.
Research project (§ 26 & § 27)
Duration : 2017-01-15 - 2020-01-14

Stress resistance and adaptation are crucial factors determining an organism’s survival to a number of biotic and a-biotic factors. The aim of this project is to elucidate the molecular basis for stress tolerance by proteomic methods and to ultimately detect proteins with key roles in counteracting stress insults. To this purpose, two strains of extremotolerant black fungi – i.e. the wild type and the natural non-melanized mutant of the species Knufia chersonesos – will be used as model organisms displaying a remarkable natural aptitude to tolerate massive ozone rates as well as protein thermo-stability. The recourse to black fungi is based on the extraordinary ability of these organisms to survive life-threatening environmental conditions, which makes them the most stress resistant Eukaryotes known to date. The two strains will enable to study the dynamics of the oxidative stress defense triggered by the exposure to ozone, in an extremotolerant species. Thus, the data obtained will allow shedding light on the oxidative stress, a condition associated with several physiological and also pathological conditions. The understanding of the basis for tolerance in adapted species will possibly aid accomplishing a second objective of the present project: the finding of novel protein candidates – e.g. radicals scavenging and antioxidant producing enzymes – with potential biotechnological and cosmetic applications. Protein stability and degradation tests will be carried out and special focus will be dedicated to enzymatic cascades reactions involved in degradation of polymers such as PBAT. Time points will be selected for the ozone treatment of the fungal strains cultivated in a multi fermenter under controlled parameters, in order to evaluate the effects of short- and long-term exposure. PBAT-mediated induction of cultures grown in liquid culture will be performed to identify polymer degrading enzymes in the extracellular medium. Shotgun quantitative proteomics approaches for simultaneous detection of changes in protein levels and identification, will be applied to the analysis of both whole-cell proteome and secretome. While the oxidative stress response will be investigated by means of thiol-based redox proteomics (cysTMTRAQ), label-free shotgun proteomics will serve to the identification of differentially expressed proteins in(un)-induced samples.
Research project (§ 26 & § 27)
Duration : 2016-12-13 - 2020-12-12

PBU, BOKU, Vienna, will provide axenic tissue cultures at different levels of organization, from shoot cultures from axillary and ad-ventitious buds, somatic embryogenesis from juvenile and adult explants, including single cell regeneration, allowing to reach the required sample size for screening within a reasonable time frame. Mutations will be induced y-ray, X-ray, EMS together with PBGL. PBU will attempt a molecular validation of mutants to provide a valid advice on how to generate, screen and recover interesting mutant genotypes of coffee (Coffea sp.) after irradiation using enabling techniques without the need for time consuming chimera dissolution processes. This might be crucial to the accelleration of success for CRP D22005.

Supervised Theses and Dissertations