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Biology subjects

Włodarczyk-Biegun, M. K.

Publications and source records attributed to Włodarczyk-Biegun, M. K..

3 recordsLinked to original sources

Melt Electrowriting of Scaffolds with a Porosity Gradient to Mimic the Matrix Structure of the Human Trabecular Meshwork

The permeability of the Human Trabecular Meshwork (HTM) regulates eye pressure via a porosity gradient across its thickness modulated by stacked layers of matrix fibrils and cells. Changes in HTM porosity are associated with increases in intraocular pressure and the progress of diseases like glaucoma. Engineered HTMs could help to understand the structure-function relation in natural tissues, and lead to new regenerative solutions. Here, melt electrowriting (MEW) is explored as a biofabrication technique to produce fibrillar, porous scaffolds that mimic the multilayer, gradient structure of native HTM. Poly(caprolactone) constructs with a height of 125-500 m and fiber diameters of 10-12 m are printed. Scaffolds with a tensile modulus between 5.6 and 13 MPa, and a static compression modulus in the range of 6-360 kPa are obtained by varying the scaffolds design, i.e., density and orientation of the fibers and number of stacked layers. Primary HTM cells attach to the scaffolds, proliferate, and form a confluent layer within 8-14 days, depending on the scaffold design. High cell viability and cell morphology close to that in the native tissue are observed. The present work demonstrates the utility of MEW to reconstruct complex morphological features of natural tissues.

bioengineering↗

Faithful Scanning Electron Microscopic (SEM) visualization of 3D printed alginate-based scaffolds

The morphological characterization of 3D printed hydrogel-based scaffolds is essential for monitoring their size, shape, surface texture and internal structure. Among other microscopic techniques, Scanning Electron Microscopy (SEM) is capable of visualizing nearly all kinds of materials at different length scales, with exceptional precision, if investigation under vacuum is possible. However, due to the high water content of hydrogel-based scaffolds and the connected volume change after drying, special preparation techniques are necessary to stabilize the 3D architecture when imaged by SEM. Here we present a straightforward cryo-SEM technique to visualize 3D printed hydrogel-based alginate scaffolds. By use of a homemade cryo-SEM holder and plunge-freezing in liquid ethane, scaffolds are visualized from the top and cross-sectional view at different magnifications. The proposed method is compared with SEM imaging in different modes (cyro-SEM, conventional SEM, ESEM) following other commonly used sample preparation techniques, such as plunging in liquid nitrogen, air-drying, freeze-drying and plunging in liquid ethane after graded dehydration. These approaches, except ESEM, lead to shrinkage, deformation, distortion or disintegration of the scaffolds and consequently give rise to artifacts in imaging. The presented results indicate that cryo-SEM after plunging in liquid ethane allows for the most faithful and time-efficient visualization of 3D printed alginate scaffolds.

bioengineering↗

Printability study of metal ion crosslinked PEG-catechol based inks

Inspired by reversible networks present in nature, we have explored the printability of catechol functionalized polyethylene glycol (PEG) based inks with metal-coordination crosslinking. Material formulations containing Al3+, Fe3+ or V3+ as crosslinking ions were tested. The printability and shape fidelity were dependent on the ink composition (metal ion type, pH, PEG molecular weight) and printing parameters (extrusion pressure and printing speed). The relaxation time, recovery rate and viscosity of the inks were analyzed in rheology studies and correlated with thermodynamic and ligand exchange kinetic constants of the dynamic bonds and the printing performance (i.e. shape fidelity of the printed structures). The relevance of the relaxation time and ligand exchange kinetics for printability was demonstrated. Cells seeded on the crosslinked materials were viable, indicating the potential of the formulations to be used as inks for cell encapsulation. The proposed dynamic ink design offers significant flexibility for 3D (bio)printing, and enables straightforward adjustment of the printable formulation to meet application-specific needs.

biochemistry↗