Biofabrication techniques to analyze and steer mechanobiology of epithelia

The motivation for this project area is to develop novel biofabrication techniques, tailored biomaterials, and methods to analyze how (stem) cells and epithelia respond to structural, physical, and mechanical properties and forces in 3D constructs and to study how their functions and behavior can be steered. The biomaterial constructs can be designed and produced to control these properties on the nm, µm, and mm scale, while they can be programmed to be dynamically altered via response to cell-induced or external triggers, such as magnetic, acoustic, or electric fields and light. Development of engineering tools that operate at the required time and length scale and allow for multimodal stimulation and measurements require know-how in mechanical, chemical, and electrical engineering. The project area bundles this type of engineering expertise and is tightly linked to the research in the other project areas in order to understand the experimental necessities and answer mechanobiological questions. Depending on the needs for each project, the building blocks can be adjusted especially with regard to stiffness, degradation kinetics, orientation, can be bioprinted into the desired architectures, and can finally be analyzed in situ in the cross-talk with the embedded cells.

Teams and Projects

DWI - Leibniz Institute for Interactive Materials

Pre-programming anisometric microgels to orthogonally study the effect of mechanical signals on epithelia in 3D tissue models

Laura De Laporte
Principal Investigator
Ramin Nasehi
Associated Postdoctoral Researcher
Iris Doolaar
Associated Doctoral Researcher
Light-responsive hydrogels to understand mechanotransduction in skeletal muscle and epithelia
Laura Klasen
Associated Doctoral Researcher
Thesis Title
Carolina Itzin
Doctoral Researcher
Pre-programming magnetic anisometric microgels to assemble into 3D macroporous regenerative structures
Project overview. (A) shows a comparison of spheroid outgrowth in PEG hydrogels (6.5% [w/w]) with different ratios of degradable crosslinkers. (B) PEG hydrogel stiffening and softening can be induced on demand with UV light. (C) Nerve growth and alignment depends on microgel stiffness in Anisogel. (D) The angle of orientation of the microgels inside an Anisogel can be preprogrammed by pre-aligning ellipsoidal maghemite nanoparticles, resulting in orthogonal alignment of microgels and cells. (E) presents RGD-functionalized ester-linked PEG microgels (8-arm, 20 kDa, 5% [w/v]) covered with immortalized CD10 kidney epithelial cells after 4 days of cultivation. (F) Degradation of ester-linked PEG microgels is observed after adding 20 mg/ml cellulase (8-arm, 20 kDa, 5% [w/v]; degradation time: 10 h).
Department of Dental Materials and Biomaterials Research (ZWBF), Uniklinik RWTH Aachen

Mechanobiological challenges related to hydrogel-based bioprinting technology for manufacturing novel 3D cell culture models

Horst Fischer
Principal Investigator
Alejandro Gómez Montoya
Associated Doctoral Researcher
Multiscale cell-preserving 3D bioprinting of human cells using the principle of nozzle-free acoustic droplet ejection
Mert Karpat
Doctoral Researcher
Mechanobiological challenges related to hydrogel-based bioprinting technology for manufacturing novel 3D cell culture models
Project overview.  (A) Collagen fibers align in response to defined dynamic stress application. (B) shows a bioreactor to investigate the effect of fluid shear stress on epithelial cells. (C) The acoustic bioprinting principle is used for realization of advanced 3D in vitro epithelial models. Single cells and cell clusters can be precisely printed in 3D and are subjected to much less shear stress during printing due to the nozzle-less technology developed in our lab.
Neuroelectronic Interfaces, RWTH Aachen University

Development of light-driven deformable and electroactive 3D scaffolds for electromechanical modulation of epidermal tissues

Francesca Santoro
Principal Investigator
Xin Yang
Doctoral Researcher
Development of a 3D oriented material construct to establish a human innervated skin disease model
Project overview. The aim is to use light-driven deformable 3D pillars (a-b) and photo-electroactive 3D pillars (c-d) to regulate keratinocyte motility, differentiation, and proliferations. (a, a’) shows poly(disperse red 1)-based light-driven deformable 3D pillars that were fabricated by means of photolithography and electron beam lithography [(a) before and (a’) after light stimulation]. (b, b’) shows fluorescence microscopy of Lifeact-RFP transfected U2OS cells growing on light-driven pillars before (b) and after light stimulation (b’). Scale bar: 10 µm. (c, c’) Shows F-actin ring-like arrangements of neurons around P3HT micropillars indicating membrane wrapping. The cells were transfected with Lifeact-RFP. Scale bar: 5 μm. (d, d’) Depicts paxillin-rich adhesions on P3HT micropillars (zoomed-in inset; green puncta). Cells were stained with TRITC-phalloidin (actin, red) and anti-paxillin antibodies (green). Scale bars: 5 μm, inset: 2 μm.