SUCCESS STORIES

Publication of Second Research Article

In October 2024, Ilias Nikolits celebrated the success of publishing his second research article, titled "Characterization of MSC Growth, Differentiation, and EV Production in CNF Hydrogels under Static and Dynamic Cultures in Hypoxic and Normoxic Conditions", in the journal MDPI Bioengineering. This publication marks another milestone in Ilias' PhD journey, bringing him closer to completing his doctoral studies at BOKU.

The peer-review process was highly constructive, with positive feedback that contributed to the refinement of an already solid piece of research. Ilias expresses excitement and pride in seeing his work recognized in the scientific community, which further supports his ongoing efforts in understanding the behavior of mesenchymal stem cells (MSCs) within cellulose-based hydrogels. The paper represents a significant contribution to the field of cell-based therapies and biomaterials, showcasing innovative approaches in advanced cell culture techniques.

With the successful publication of this article, Ilias is not only closer to achieving his PhD but also excited for the opportunities this research will open up in the future.

In the quest to improve nerve regeneration for patients suffering from injuries to the peripheral nervous system, spider silk has emerged as a promising material. This remarkable natural fiber can be used as a filler in nerve guidance conduits, helping nerves regenerate more effectively. Imagine a silk-filled implant placed in the body between two damaged nerve ends, supporting the body’s natural healing process.

The goal of my research is to understand the structure-function relationships of natural spider silk, which is key to producing it artificially for medical use. To do this, I’m investigating how mechanical stress and strain impact the silk’s inner crystal structure.

One of the most exciting experiments we conducted involved testing the silk fibers under mechanical strain using X-rays. This was done at two world-leading synchrotron facilities—MAX IV in Sweden and the ESRF in France. By stretching single spider silk fibers while exposing them to a focused X-ray beam, we were able to observe how the structure inside the silk changes under tension.

This was no easy task! The X-ray beam, with a focus as small as 100 nanometers, is so powerful that it can damage the delicate silk fibers. We had to carefully adjust our scanning parameters to avoid cutting the fibers with the beam. Nevertheless, we successfully tested more than 100 individual silk fibers (including several night shifts), both in ambient conditions and at varying levels of humidity.

The results of this study bring us one step closer to understanding the unique properties of spider silk. By unlocking these secrets, we hope to design artificial silk that can improve the success of nerve regeneration in the future—giving patients better outcomes and reducing suffering.

In September 2024, Elettra Verin and Oscar García attended to the 5^th European summer school on microfluidics at the University “Magna Graecia” in Catanzaro, Italy. The training strengthened their knowledge on biomaterials for cell culture, and expanded their horizons on microfabrication, in-line analytics, organs on a chip, visualization and cell therapy in microfluidic devices, among other selected topics from experts on the field. The summer school also offered hands-on sessions and several networking opportunities with peers and industry partners.

Elettra, who recently joined our doctoral school, presented her project “Optimization of a 3D analysis platform for the assessment of cartilage graft maturation in vitro”, which provides an alternative to animal testing for cartilage regeneration therapies. Her project uses concepts from miniaturization and in-line sensors, developing a bioreactor platform to timely provide information on the maturity of developing cartilage grafts, sampling different metabolites from within and outside the cultured tissue. Oscar, who is on his third year at our doctoral school, presented the results of his project “Establishing a high throughput fluidic system for mesenchymal stem cells encapsulation and expansion”, which enables the generation of plentiful low cost and high fidelity microgels, to observe proliferation and metabolic activity of adherent cells in dynamic culturing systems. His technology has been successfully applied to stem cell differentiation and manufacture of extracellular vesicles.

“We are grateful that the doctoral school gave us such an opportunity, which for sure  contributes to the accomplishment of our scientific goals and connects us with our research community, we also encourage our colleagues to join to the next instalment of the microfluidics summer school which will be held in Denmark in 2025”.

Julia Moldaschl was honored to be accepted for a flash talk and poster presentation at the 16th ÖGMBT Annual Meeting in Graz, held on 17th September 2024. She was thrilled by the opportunity to participate in such a renowned event and looked forward to sharing her latest research and connecting with fellow scientists. However, due to unforeseen weather conditions and train delays caused by heavy rain, Julia was unable to arrive in time for the flash talk. Despite this unexpected challenge, the day turned out to be incredibly rewarding.

Julia fully embraced the opportunity to present her research during the poster session, where she had lively and enriching discussions with other scientists. Her poster, titled „Spheroid Trilineage Differentiation Model of Primary Mesenchymal Stem/Stromal Cells under Hypoxia and Serum-Free Culture Conditions” sparked meaningful exchanges, and she connected with new colleagues in the field – as visible on the picture.

Although the journey did not go as planned, the day still proved to be a fantastic success, filled with collaboration, inspiration, and fresh perspectives for the future.

Here are some highlights of my experience at the RÅC International Summer School 2023, which took place from 20 to 27 August 2023 in Lüneburg, Germany. The central theme of this event was "Cutting-edge Neutron and X-ray Research for a Sustainable Future". During my time at the Summer School, I had the privilege of presenting a poster entitled "Nanobeam X-ray Diffraction on Various Spider Silk Fibres". I am particularly grateful for the opportunity to interact with fellow participants from different parts of the world. To further enrich my participation, I took part in a Science Slam presentation where I shared my insights on spider silk. The overwhelmingly positive response from the audience was truly inspiring. In addition to my own contributions, the Summer School provided an invaluable opportunity to attend a series of cutting-edge scientific lectures by renowned scientists in their respective fields from Germany and Sweden. The Summer School broadened my intellectual horizons, rekindled my passion for materials science and strengthened my commitment to sustainable solutions.

“Life Sciences and cutting-edge technologies” was this year’s topic of the annual meeting (2023) of the Austrian Association of Molecular Life Sciences and Biotechnology (OEGMBT), which took place from 19^th-21^st of September in Salzburg, Austria. A broad palette of topics was covered, which included current developments in applied and medical life sciences. In this context, also basic research was represented by a diverse set of talks.  The organizers struck a good balance between internationally renowned speakers and young researchers, allowing them to showcase their work in front of a large crowd. Of the 62 high-quality oral lectures, 2 were given by DocSchool students (Julia Moldaschl, Ilias Nikolits) and 3 of the 71 posters came from students of the BioMatInt program. In the final award ceremony, the poster presentation by Sabrina Nebel was awarded one of the three prizes for an outstanding poster presentation kindly sponsored by The FEBS Journal. We congratulate!

Further information
https://febs.onlinelibrary.wiley.com/hub/journal/17424658/features/the-febs-journal-poster-prize-

The Article "Alginate Core-Shell Capsules for 3D Cultivation of Adipose-Derived Mesenchymal Stem Cells”, published in February 2022, was recently selected as "Editor's Choice". The paper was part of the Special Issue “Material and Engineering-Based Approaches for Organoids” in the Open Access journal MDPI Bioengineering and was the first lead-authorship publication of Sabrina Nebel, doctoral candidate at the Institute of Cell and Tissue Culture Technologies.

Sabrina’s research is focused on mesenchymal stem cells (MSCs), progenitor cells of the connective tissue that can be found in almost all tissues, even in adults. These cells are easily available and can be used in a wide variety of cell therapies and in-vitro testing systems. However, since millions of cells are required for any application, ex-vivo cell expansion is inevitable. Cultivation in petri dishes in 2D is still considered the gold standard, despite evidence that 3D cultivation not only retains but can even improve the beneficial properties of the cells.

Ilias Nikolits gave his first conference talk at the 15th ÖGMBT Annual Meeting in September 2023, held in Salzburg, Austria. As part of the "Regenerative Medicine & Stem Cells" session, Ilias presented his research titled, "Effects of different in vitro cultivation conditions on human mesenchymal stem cells encapsulated in TEMPO-oxidized nanofibrillated cellulose hydrogels." His talk focused on recent findings from his PhD work, which explores how varying cultivation environments impact MSC growth, differentiation, and extracellular vesicle production in cellulose-based hydrogels. This study is particularly relevant for improving cell-based therapies and in vitro testing approaches.

“This was my first experience presenting at an international conference, and it was both exciting and rewarding. I felt a great sense of pride in being able to share my findings with leading experts in the field. Engaging with fellow researchers and receiving feedback was incredibly valuable, and it has inspired me to continue pushing the boundaries of my research. Overall, the experience strengthened my confidence in public speaking and allowed me to showcase the progress and potential impact of my work.”

Julia Moldaschl is a doctoral candidate at the Institute of Cell and Tissue Culture Technologies (ICTCT) at the Department of Biotechnology since September 2021. One of her most exciting achievements so far was her contribution to the 14^th ÖGMBT Annual Meeting, in Vienna in September 2022. She gave a talk titled “Multilineage Differentiation of Juvenile Primary Bone Marrow-Derived Mesenchymal Stem Cells in 3D under Normoxic and Physioxic Conditions” in the section “Cell Based Assays, Therapies and Products”. This posed an ideal opportunity for Julia to share her experience with mesenchymal stem/stromal cells (MSCs) cultured and differentiated under physiological conditions.

In particular, her research focuses on the establishment of a MSC multilineage differentiation model under physiological conditions, which comprise scaffold-free 3D culture approaches, reduced oxygen conditions, that can also be considered as “Physioxia”, and the application of xeno- and serum-free media.

Poster Prize – ÖGMBT Workshop 2023

On 17^th March 2023 Julia attended the ÖGMBT Workshop “Advanced Cell Culture Technologies” and was awarded the prize for the best poster. The workshop was organized by the ÖGMBT working group “Cell based assays, therapies and products” and took place directly at BOKU. During the workshop Julia joined lively discussions, exchanged experience in 3D cell culture technologies and connected with new colleagues. “It is such an enriching experience! I got helpful feedback on my work and so many questions that benefited my thesis writing”. On top of this exciting day, Julia won the poster prize for her poster entitled “3D Multilineage Differentiation Model of Mesenchymal Stem/Stromal Cells under Physiological Conditions”.

Karolina Peter, member of the Doctoral School Biomaterials and Biointerfaces, could engage many interested listeners with her scientific presentation at the JUNIOR EUROMAT conference (19-22 July, Coimbra, Portugal). The presentation, titled "Zooming inside spider silk" - Using synchrotron nanoXRD to compare the ultrastructure of spider silks." was very well received and motivated many of the audience to ask interested questions. There were research results presented, which were collected at the Institute of Physics and Materials Science (BOKU).

Moreover, this was the first conference for Karolina that was not held in a virtual setting, and her first oral presentation.In addition to numerous other interesting lectures, one could also participate in workshops such as Electron Microscopy or Horizon Europe. In particular, the contact with other young scientists was great at JUNIOR EUROMAT 2022 in Coimbra. All in all, a very successful experience, both personally and scientifically valuable.

Everything started with a remote experiment at the end of 2020, to which the team from the Institute of Physics and Material Science (IPM) was unfortunately unable to go in person due to Covid-19. Then, in June 2022, the time had finally come, and it was possible to travel in person to the ESRF (European Synchrotron and Radiation Facility) in Grenoble, France. The proposal for an experiment on spider silk was accepted, which is reason to celebrate.

While I boarded my flight from Vienna to Copenhagen on March 1^st, I knew that I would get to meet one of the most renowned professors for computer simulation of intrinsically disordered proteins in Europe. However, I did not expect to also find a group of great colleagues and a progressive and beautiful city to live in. There is a lot to learn from the city of Copenhagen, especially how to build a top-notch biking infrastructure; the best I have ever seen in my life.

I was able to significantly progress my PhD project, specifically in regard to ensemble reweighting; a topic the Lindorff-Larsen research group has vast experience in. I was given time to think through the mathematical details of their method and to experiment with their data.

During my stay in Copenhagen, I learned:

* … how to use the BME-Reweighting tool.
* … the workflow of a RNA reweighting-example.
* … insight into methodology and mathematical background of the BME method.
* … insight into danish culture and a (little) bit of their language

After visiting Scandinavia in the northern part of Europe, I had the chance to join a one-week long seminar about disordered proteins at Institut d'Etudes Scientifiques de Cargèse. Located on the island of Corsica, France, it was an adventure to get there with in-between stops in Paris, Nice and Ajaccio. The seminar was hold at a very remote location directly on the shore of the Mediterranean Sea. I was able to meet more than 30 like-minded colleagues from Europe, the US and from Japan. Lectures and trainings where held by senior scientists from France, Germany and the United States.

While joining the seminar, I learned:

* … how to use an ensemble refinement method.
* … how to use Flexible-Meccano to generate conformational ensembles.
* … how to approach IDP phase-separation simulation.

I’m very glad that BioMatInt gave me opportunity to take part in those events and I look forward to an equally successful 2023.

In October 2022, Goodness Osondu-Chuka won an attendance grant to participate in and present her Ph.D. work at the ESCMID/ASM Conference on Drug Development to Meet the Challenge of Antimicrobial Resistance (2022), which took place in Dublin, Ireland.

Goodness is a doctoral candidate of the BioMatInt at the Institute of Biologically Inspired Materials. In her project, she creates a three-dimensional biofilm model that replicates key aspects of the biofilms found in the lungs of individuals with cystic fibrosis, a chronic illness without a cure. Through this model, she intends to investigate and comprehend the protective functions of biofilms in persistent infections, particularly in the lungs of cystic fibrosis patients, that prevent effective antibiotic treatments.

She applied for this grant because it offered an excellent opportunity to share and gain more knowledge on various aspects of microbial biofilms as well as novel techniques and approaches for solving the problem of antimicrobial resistance and tolerance in chronic infections. The grant allowed Goodness to present her first results on extracting, reconstituting, and printing the extracellular polymers that provide the structural properties of cystic fibrosis biofilms.

The conference, which was focused on the challenges of developing new agents and solutions for antimicrobial resistance, started with a half-day boot camp designed to share the wealth of experience from the community. This was followed by two and a half days of high-quality symposia, roundtable discussions, keynote, and young investigator lectures. The meeting was designed to bring together researchers, academics, industry regulators, and funders to share experiences, collaborate and learn. In her own words, “Attending the conference was a fantastic chance to stay informed about current research, express my thoughts, and network effectively. I eagerly anticipate similar opportunities to attend and actively participate in future conferences”. 

The ESEM has not been used for the last two years. I had to completely skip and clean the water cooling to get the brown algae out of the pipes. But the result is very satisfying, you can examine a fresh organic sample without much drying effects in environmental mode with water vapour at 400 Pascal pressure. Between the sample, cooled to 4 °C, and the electron beam outlet are water molecules that protect the sample from drying out and also guide the secondary electrons to the detector. In the pictures you can see the setup in the chamber of the ESEM and in the micrograph a vacuole filled with essential oil in the outer shell of a blood orange. 

First lead authorship publication

On February 2023 the article “Hydrogels from TEMPO-Oxidized Nanofibrillated Cellulose Support In Vitro Cultivation of Encapsulated Human Mesenchymal Stem Cells” was published on the ACS Applied Bio Materials journal. This was the first lead authorship publication of Ilias Nikolits, doctoral candidate at the Institute of Cell and Tissue Culture Technologies (ICTCT) at the Department of Biotechnology of BOKU.

The aim of Ilias’ PhD project is on the development of hydrogels for 3D in vitro cultivation of human mesenchymal stem cells (MSCs). The material he is focusing on his research is cellulose, a highly abundant and inexpensive natural polymer, with high biocompatibility and tunable properties. The hydrogel scaffolds synthesized from cellulose are aiming to provide a 3D in vivo-like environment during cultivation of MSCs in vitro, compared to the gold standard 2D plastic adherent methods, by preserving the physiological characteristics and functions of the cells.

In this publication, Ilias shows that hydrogels synthesized from chemically functionalized cellulose nanofibers consist a very promising platform for in vitro cultivation of humans MSCs. Encapsulation of MSCs in those hydrogels mimics a 3D tissue-like environment and cells can perform their physiological functions and behavior.

“When people ask me what hydrogels are, I give them the example of Gummy Bears. So, I am basically putting stem cells in Gummy Bears made from wood powder. But is seems the cells really like it” -Ilias quotes.

Research stay abroad - Magnetically triggered drug nanocapsules in hydrogels

University of Siegen, Germany (February 2023) 

In February 2023, Giacomo Chizzola spent a month at the University of Siegen, Germany, to expand his knowledge and abilities in polymer synthesis. Giacomo, a member of the Biomaterials and Biointerfaces Doctoral School, is a doctoral candidate at the Institute of Biologically Inspired Materials.