Extracellular Biotechnology

In the Extracellular Biotechnology research group, expertise from tissue regeneration, ageing, vascular biology, and nanoparticle-based therapies comes together. The shared goal is to better understand the biological, cellular, and physical foundations of tissue healing across the lifespan – and to translate these insights into diagnostic and therapeutic strategies.

Tissue Regeneration and Ageing

As the body ages, its ability to regenerate organs and tissues declines significantly. The first signs of reduced resilience following traumatic injury can already be observed between the ages of 45 and 50. A key contributor to this decline is the accumulation of senescent cells, whose senescence-associated secretory phenotype (SASP) promotes inflammation and inhibits the body’s natural repair mechanisms.

A central focus of the group is to investigate the role of microRNAs (miRNAs) and differentially regulated proteins in senescent and stressed cells, and how these affect tissue regeneration and organ ageing. By analysing tissue models and studying extracellular vesicles (EVs), the team aims to identify novel senolytic targets and therapeutics that selectively combat cellular senescence and enhance regenerative capacity. Particular attention is given to the use of mesenchymal stem cell (MSC)-derived EVs as a platform technology to support the healing of bone, cartilage, skin, and neural tissue in both young and ageing organisms.

Vascular Biology

A functioning blood and lymphatic supply is fundamental to any form of tissue regeneration. Endothelial cells, which line the vascular system, are essential in this context: they not only regulate blood flow but also actively participate in processes such as angiogenesis, the formation of new blood vessels.

The group therefore also focuses on the role of endothelial cells in the formation and growth of blood and lymphatic vessels. This includes co-cultures of endothelial cells and mesenchymal stem cells, which are applied in tissue engineering to enhance the vascularisation of implanted tissues. In addition, the role of EVs and purinergic signalling pathways in angiogenesis is being explored.

The objective is to develop clinically applicable vascularisation strategies, with an emphasis on the characterisation and development of artificially engineered vascular structures, as well as the regeneration of existing blood and lymphatic vessels.

Nanoparticle-Based Therapies

In addition to biological research, the group pursues a strong technological focus: the development of methods for the quantitative analysis of biological systems at the nanoscale. This includes extensive experience in single-molecule fluorescence microscopy, atomic force microscopy, and 2D/3D nanolithography.

By precisely locating and measuring the dynamics of biomolecules, and by determining molecular interactions, the group seeks to gain deeper insight into molecular function in both model systems and living cells. Current research centres on the development of methods to characterise and understand the physical properties of individual cells, nanobioparticles, lipids, and proteins.

Another key aspect is the translation of these findings into new device technologies and imaging tools.

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Selected Publications

Bobbili MR, Görgens A, Yan Y, Vogt S, Gupta D, Corso G, Barbaria S, Patrioli C, Weilner S, Pultar M, Jacak J, Hackl M, Schosserer M, Grillari R, Kjems J, Andaloussi SE, Grillari J (2024). Snorkel-tag based affinity chromatography for recombinant extracellular vesicle purification. J Extracell Vesicles. 2024 Oct;13(10):e12523. 
(free PDF)

Lushchak O, Schosserer M, Grillari J. Senopathies-Diseases Associated with Cellular Senescence (2023). Biomolecules. 2023 Jun 8;13(6):966. 
(free PDF)

Messner Z, Carro-Vazquez D, Haschka J, Grillari J, Resch H, Muschitz C, Pietschmann P, Zwerina J, Hackl M, Kocijan R (2022). Circulating miRNAs Respond to Denosumab Treatment after Two Years in Postmenopausal Women with Osteoporosis. J Clin Endocrinol Metab. 2022 Nov 19:dgac667. 

Schanda JE, Heher P, Weigl M, Drechsler S, Schädl B, Prueller J, Kocijan R, Heuberer PR, Hackl M, Muschitz C, Grillari J, Redl H, Feichtinger X, Fialka C, Mittermayr R (2022). Muscle-Specific Micro-Ribonucleic Acids miR-1-3p, miR-133a-3p, and miR-133b Reflect Muscle Regeneration After Single-Dose Zoledronic Acid Following Rotator Cuff Repair in a Rodent Chronic Defect Model. Am J Sports Med. 2022 Oct;50(12):3355-3367. 

Carro Vázquez D, Emini L, Rauner M, Hofbauer C, Grillari J, Diendorfer AB, Eastell R, Hofbauer LC, Hackl M (2022) Effect of Anti-Osteoporotic Treatments on Circulating and Bone MicroRNA Patterns in Osteopenic ZDF Rats. Int J Mol Sci. 2022 Jun 10;23(12):6534. 
(free PDF).

Muschitz C, Hummer M, Grillari J, Hlava A, Birnger AH, Hemetsberger M, Bimai HP (2022) Epidemiology and economic burden of fragility fractures in Austria. Osteoporos Int 2022. Mar;33(3):637-647. 
(free PDF)

Zupan J, Strazar K, Kocijan R, Nau T, Grillari J, Marolt Presen D (2021) Age-related alterations and senescence of mesenchymal stromal cells: Implications for regenerative treatments of bones and joints. Mech Ageing Dev 2021 Sep;198:111539. 

Vogt S, Bobbili MR, Stadlmayr G, Stadlbauer K, Kjems J, Rüker F, Grillari J, Wozniak-Knopp (2021). An engineered CD81-based combinatorial library for selecting recombinant binders to cell surface proteins: Laminin binding CD81 enhances cellular uptake of extracellular vesicles. J Extracell Vesicles. 2021 Sep;10(11):e12139.
(free PDF)

Pils V, Terlecki-Zaniewicz L, Schosserer M, Grillari J, Lämmermann I (2021) The role of lipid-based signalling in wound healing and senescence. Mech Ageing Dev2021 Sep;198:111527.

Kocijan R, Weigl M, Skalicky S, Geiger E, Ferguson J, Leinfellner G, Heimel P, Pietschmann P, Grillari J, Redl H, Hackl M (2020) MicroRNA levels in bone and blood change during bisphosphonate and teriparatide therapy in an animal model of postmenopausal osteoporosis. Bone. 2020 Feb;131:115104.

Hauser F, Naderer C, Priglinger E, Peterbauer A, Fischer MB, Redl H, Jacak J (2024). Single molecule studies of dynamic platelet interactions with endothelial cells. Front Bioeng Biotechnol. 2024 Apr 3;12:1372807. 
(free PDF)

Sivun D, Murtezi E, Karimian T, Hurab K, Marefat M, Klimareva E, Naderer C, Buchroithner B, Klar TA, Gvindzhiliia G, Horner A, Jacak J (2024). Multiphoton lithography with protein photoresists. Mater Today Bio. 2024 Feb 10;25:100994. 
(free PDF)

Sych T, Schlegel J, Barriga HMG, Ojansivu M, Hanke L, Weber F, Beklem Bostancioglu R, Ezzat K, Stangl H, Plochberger B, Laurencikiene J, El Andaloussi S, Fürth D, Stevens MM, Sezgin E. (2023). High-throughput measurement of the content and properties of nano-sized bioparticles with single-particle profiler. Nat Biotechnol. 2023 Jun 12. 

Puthukodan S, Hofmann M, Mairhofer M, Janout H, Schurr J, Hauser F, Naderer C, Preiner J, Winkler S, Sivun D, Jacak J (2023). Purification Analysis, Intracellular Tracking, and Colocalization of Extracellular Vesicles Using Atomic Force and 3D Single-Molecule Localization Microscopy. Anal Chem. 2023 Apr 11;95(14):6061-6070. 
(free PDF)

Strohmeier K, Hofmann M, Jacak J, Narzt MS, Wahlmueller M, Mairhofer M, Schaedl B, Holnthoner W, Barsch M, Sandhofer M, Wolbank S, Priglinger E. (2023). Multi-Level Analysis of Adipose Tissue Reveals the Relevance of Perivascular Subpopulations and an Increased Endothelial Permeability in Early-Stage Lipedema. Biomedicines. 2022 May 18;10(5):1163.
(free PDF)

Puthukodan S, Hofmann M, Mairhofer M, Janout H, Schurr J, Hauser F, Naderer C, Preiner J, Winkler S, Sivun D, Jacak J. (2023). Purification Analysis, Intracellular Tracking, and Colocalization of Extracellular Vesicles Using Atomic Force and 3D Single-Molecule Localization Microscopy. Anal Chem. 2023 Apr 11;95(14):6061-6070. 
(free PDF)

Strohmeier K, Hofmann M, Hauser F, Sivun D, Puthukodan S, Karner A, Sandner G, Le Renard PE, Jacak J, Mairhofer M (2021). CRISPR/Cas9 Genome Editing vs. Over-Expression for Fluorescent Extracellular Vesicle-Labeling: A Quantitative Analysis. Int J Mol Sci. 2021 Dec 28;23(1):282. 
(free PDF)

Priglinger E, Strasser J, Buchroithner B, Weber F, Wolbank S, Auer D, Grasmann E, Arzt C, Sivun D, Grillari J, Jacak J, Preiner J, Gimona M (2021). Label-free characterization of an extracellular vesicle-based therapeutic. J Extracell Vesicles. 2021 Oct;10(12):e12156.
(free PDF)

Crnic A, Rohringer S, Tyschuk T, Holnthoner W (2024). Engineering blood and lymphatic microvascular networks. Atherosclerosis. 2024 Jun;393:117458. 
(free PDF)

Pultar M, Oesterreicher J, Hartmann J, Weigl M, Diendorfer A, Schimek K, Schädl B, Heuser T, Brandstetter M, Grillari J, Sykacek P, Hackl M, Holnthoner W (2024). Analysis of extracellular vesicle microRNA profiles reveals distinct blood and lymphatic endothelial cell origins. J Extracell Biol. 2024 Jan 15;3(1):e134. 
(free PDF)

Hromada C, Hartmann J, Oesterreicher J, Stoiber A, Daerr A, Schädl B, Priglinger E, Teuschl-Woller AH, Holnthoner W, Heinzel J, Hercher D (2022). Occurrence of Lymphangiogenesis in Peripheral Nerve Autografts Contrasts Schwann Cell-Induced Apoptosis of Lymphatic Endothelial Cells In Vitro. Biomolecules. 2022 Jun 12;12(6):820. 
(free PDF).

Strohmeier K, Hofmann M, Jacak J, Narzt MS, Wahlmueller M, Mairhofer M, Schaedl B, Holnthoner W, Barsch M, Sandhofer M, Wolbank S, Priglinger E (2022). Multi-Level Analysis of Adipose Tissue Reveals the Relevance of Perivascular Subpopulations and an Increased Endothelial Permeability in Early-Stage Lipedema. Biomedicines. 2022 May 18;10(5):1163. 
(free PDF)

Trisko J, Fleck J, Kau S, Oesterreicher J, Holnthoner W (2022). Lymphatic and Blood Endothelial Extracellular Vesicles: A Story Yet to Be Written. Life (Basel). 2022 Apr 28;12(5):654. 
(free PDF)

Schneider J, Pultar M, Oesterreicher J, Bobbili MR, Mühleder S, Priglinger E, Redl H, Spittler A, Grillari J, Holnthoner W (2021) Cre mRNA Is Not Transferred by EVs from Endothelial and Adipose-Derived Stromal/Stem Cells during Vascular Network Formation. Int J Mol Sci 2021 Apr 14;22(8):4050
(free PDF)

Oesterreicher J, Pultar M, Schneider J, Mühleder S, Zipperle J, Grillari J, Holnthoner W (2021). Fluorescence-Based Nanoparticle Tracking Analysis and Flow Cytometry for Characterization of Endothelial Extracellular Vesicle Release. Int J Mol Sci. 2020 Dec 4;21(23):9278.
(free PDF)

Mühleder S, Fuchs C, Basílio J, Szwarc D, Pill K, Labuda K, Slezak P, Siehs C, Pröll J, Priglinger E, Hoffmann C, Junger WG, Redl H, Holnthoner W (2019). Purinergic P2Y2 receptors modulate endothelial sprouting. Cell Mol Life Sci. 2019 Jul 5.

Nürnberger S, Lindner C, Maier J, Strohmeier K, Wurzer C, Slezak P, Suessner S, Holnthoner W, Redl H, Wolbank S, Priglinger E (2019). Adipose-tissue-derived therapeutic cells in their natural environment as an autologous cell therapy strategy: the microtissue-stromal vascular fraction. Eur Cell Mater. 2019 Feb 22;37:113-133.

Pill K, Melke J, Mühleder S, Pultar M, Rohringer S, Priglinger E, Redl HR, Hofmann S, Holnthoner W (2018). Microvascular Networks From Endothelial Cells and Mesenchymal Stromal Cells From Adipose Tissue and Bone Marrow: A Comparison. Front Bioeng Biotechnol. 2018 Oct 25;6:156.
(free PDF)

Holnthoner W, Bonstingl C, Hromada C, Muehleder S, Zipperle J, Stojkovic S, Redl H, Wojta J, Schoechl H, Grillari J, Weilner S, Schlimp C. (2017). Endothelial Cell-derived Extracellular Vesicles Size-dependently Exert Procoagulant Activity Detected by Thromboelastometry. SciRep 25th April 2017 : 3707 (2017)
(free PDF)

Hromada C, Muehleder S, Grillari J, Redl H, Holnthoner W. (2017). Endothelial Extracellular Vesicles – Promises and Challenges. Front Physiol. 2016; doi: 10.3389/fphys.2017.00275
(free PDF)

Knezevic L, Schaupper M, Mühleder S, Schimek K, Hasenberg T, Marx U, Priglinger E, Redl H, Holnthoner W. (2017). Engineering Blood and Lymphatic Microvascular Networks in Fibrin Matrices. Front. Bioeng. Biotechnol. 5:25. doi: 10.3389/fbioe.2017.00025
(free PDF)

Schaupper MV, Jeltsch M, Rohringer S, Redl H, & Holnthoner W. (2016). Lymphatic vessels in Regenerative Medicine and Tissue Engineering. Tissue Eng Part B Rev, 22(5):395-407.

Schneider K, Aigner P, Monforte X, Nuernberger S, Holnthoner W, Ruenzler D, Redl H, & Teuschl AH. (2016). Decellularized human placenta chorion matrix as a favorable source of small diameter vascular grafts. Acta Biomaterialia, 29:125-34.

Pill K, Hofmann S, Redl H, & Holnthoner W. (2015). Vascularization mediated by mesenchymal stem cells from bone marrow and adipose tissue: a comparison. Cell Regen (Lond), 4:8.
(free PDF)