Wychowaniec, J. K. | Search bioRxiv

bioRxiv2025

3D Bioprinted Cell-laden GrooveNeuroTube: A Multifunctional Platform for Ex Vivo Neural Cell Migration and Growth Studies

Extensive peripheral nerve injuries often lead to the loss of neurological function due to slow regeneration and limited recovery over large gaps. Current clinical interventions, such as nerve guidance conduits (NGCs), face challenges in creating biomimetic microenvironments that effectively support nerve repair. The developed GrooveNeuroTube is composed of hyaluronic acid methacrylate and gelatin methacrylate hydrogel, incorporating active agents (growth factors and antibacterial agents) encapsulated within an NGC conduit made of 3D-printed PCL grid fibers. In vitro studies showed that GrooveNeuroTube significantly promoted migration of dorsal root ganglion (DRG) neuronal cells, 3D bioprinted at the far ends of the conduit to imitate a proximal nerve injury as a novel ex vivo model. A long-term culture of up to 60 days was employed to better mimic in vivo conditions. This model tested the effects of pulsed electromagnetic field (PEMF) stimulation on neural tissue development. After 60 days, GrooveNeuroTube showed a 32% cell migration increase compared to the growth-factor-group and 105% compared to the no-growth-factor condition. These results confirm that the GrooveNeuroTube system can effectively support sustained neural cell migration and maturation over extended periods, proving a new technology for testing peripheral nerve injury ex vivo. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=77 SRC="FIGDIR/small/639097v2_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@14c7799org.highwire.dtl.DTLVardef@14011c3org.highwire.dtl.DTLVardef@14e5a79org.highwire.dtl.DTLVardef@115e281_HPS_FORMAT_FIGEXP M_FIG C_FIG The graphical abstract was created with BioRender.com.

bioengineering↗

bioRxiv2024

Effect of molecular weight of tyramine-modified hyaluronan on polarization state of peripheral blood mononuclear cells-derived macrophages

The immunomodulatory properties of hyaluronan and its derivatives are key to their use in medicine and tissue engineering. In this work we evaluated the capability of soluble tyramine-modified hyaluronan (THA) of two molecular weights (low Mw=280 kDa and high Mw=1640 kDa) for polarization of THP-1 and peripheral blood mononuclear cells (PBMCs)-derived macrophages (M{Phi}s). We demonstrate the polarization effects of the supplemented THA by flow cytometry and multiplex ELISA for the THP-1 derived M{Phi}s and by semi-automated image analysis from confocal microscopy, immunofluorescent staining utilising CD68 and CD206 surface markers, RT-qPCR gene expression analysis, as well as using the enzyme-linked immunosorbent assay (ELISA) for PBMCs-derived M{Phi}s. Our data indicate that supplementation with LMW THA drives changes in THP-1 derived M{Phi}s towards a pro-inflammatory M1-like phenotype, whereas supplementation with the HMW THA leads to a more mixed profile with some features of both M1 and M2 phenotypes, suggesting either a heterogeneous population or a transitional state. For cells directly sourced from human patients, PMBCs-derived M{Phi}s, results exhibit a higher degree of variability, pointing out a differential regulation of factors including IL-10 and CD206 between the two cell sources. While human primary cells add to the clinical relevance, donor diversity introduces wider variability in the dataset, preventing drawing strong conclusions. Nevertheless, the M{Phi}s profiles observed in THP-1 derived cells for treatments with LMW and HMW THA are generally consistent with what might be expected for the treatment with non-modified hyaluronans of respective molecular weights, confirming the known association holds true for the chemically tyramine-modified hyaluronan. We stipulate that these responses will provide basis for more accurate in vivo representation and translational immunomodulatory guidance for the use of THA-based biomaterials to a wider biomaterials and tissue engineering communities. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=114 SRC="FIGDIR/small/575241v3_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@899d95org.highwire.dtl.DTLVardef@c8b80aorg.highwire.dtl.DTLVardef@1aad98dorg.highwire.dtl.DTLVardef@1b0ce60_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗