R&D inno Gewebe


R&D-Project 'Multimodal Tissue Models for preclinical testing of medicinal Products and Drugs'

ZIM-Coollaborative Project (funded by the federal Ministry of Economy and Energy)

Within the framework of the ZIM collaborative network project, an artificial tissue platform based on cell-seeded bacterial nanocellulose for modeling epithelial organ barriers (skin, lung, gut) shall be developed and tested. Tests include monitoring the effects of drugs on the epithelial layers of these organ systems. So far used animal models may suffer from limited transferability and intensive costs. Thus, our model will also contribute to reducing animal tests for drug screening.

In our project, established human epithelial cell lines (Caco-2, Calu-3, HaCaT) are used to build our in vitro tissue models for subsequent drug application and assessment of barrier function and intactness. Initially, PET membranes are cell-seeded (hanging insert systems with capillary-pore membranes), and later extended towards biotechnologically produced bacterial nanocellulose (BNC) fleeces inserted in cell crests. BNC is a very promising novel advanced hydropolymer material for tissue engineering. BNC-fleeces with variable micro-structured and functionalised surface properties are available through the commercial project partner JeNaCell (Fig. 1).

The effects of selected toxic (corrosive/irritative) drugs will be tested on the cell-scaffold-tissue barrier using microscopy technologies, in particular multiphoton excitation for minimally invasive online monitoring of the tissue and production of extracellular matrix (i.e. collagen-I). (Immuno-)fluorescence microscopy complements these analyses with respect to live/dead distributions of cells (calcein AM/ethidium-homodimer) and tight junction modulation. Moreover, additional assays to determine cell viability and activity will be employed (e.g. MTS, WST-1). From these analyses, dose-response curves for tested substances will be reconstructed. Our project idea follows the aspect of optical validation pipelines for cell and tissue models into BNC fleeces on the one hand and the direct fluorescence-based validation of cellular fitness, on the other hand.

Some project partners within the consortium (3 SMEs, 2 R&D-partners) will assess substance effects regarding their influence on barrier integrity/tightness by impedance and TER-recordings) and using multimodal micro-senors to obtsain environmental profiles on O2, CO2, pH. The miniaturised multimodal sensor elements from the SME partner Presens (Fig. 1) is extended far beyond sensor arrays available in existing commercially available in vitro test systems. The tissue models will be acclimatised within a micro-climate chamber.

The obtained drug effect endpoint avlues (regarding toxicity, corrosiveness, etc.) will be implemented into simulation models to predict drug toxicity and to extract dose-response parameters. Compounds used are stadnard substances with an established toxicity profile according to 'OECD guidelines for the testing of chemicals' from RHE-models (reconstructed human epidermis).

One of the cell models we employ is the Caco-2 cell model which is an established in vitro model for healthy human gut epithelium. It exerts excellent barrier functions and grows in well differentiated monolayers with clear polarisation.

Project partners:

Figure 1: Overview of the project partners within the ZIM consortium "innoGewebe" including 3 SMEs (JeNaCell, Presens, senetics healthcare) and two university R&D-institutions (MBT, TERM).

funded by:

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Literature:

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Abitbol T, Rivkin A, Cao Y, Nevo Y, Abraham E, Ben-Shalom T, Lapidot S, Shoseyov O. Nanocellulose, a tiny fiber with huge applications. Curr Opin Biotechnol. 2016 Jun;39:76-88. doi: 10.1016/j.copbio.2016.01.002. Review.

Dugan JM, Gough JE, Eichhorn SJ. Bacterial cellulose scaffolds and cellulose nanowhiskers for tissue engineering. Nanomedicine (Lond). 2013 Feb;8(2):287-98. doi: 10.2217/nnm.12.211. Review.