A translational approach integrating developmental biology and tissue

Background

Intervertebral disc (IVD) degeneration can lead to chronic low back pain, which is a leading cause of disability worldwide. IVD degeneration can be characterized by the dehydration of the central nucleus pulposus (NP) and subsequent structural breakdown of the peripheral annulus fibrosus (AF). The current state of AF tissue engineering could be advanced with a strategy that captures the essential building blocks of AF tissue morphogenesis: Biomimetic cell patterning and mechanical stimulation to induce circumferential cell elongation.

Goal

1) Development of a free-form reversible embedding technique that supports the bioprinting of cells and cell-laden bioinks into biomimetic patterns within three-dimensional hydrogel matrices, and 2) Exploration of cell-scale and macro-scale mechanical stimulations on cytoskeletal organization and AF collagen assembly by the patterned cells.

Results

We have developed a cell-friendly hydrogel support bath comprised of temperature-sensitive poly(N-isopropylacrylamide) hydrogels and embedded Carbopol® microparticles. The hydrogel support baths possess flow points and self-healing rheological behaviors that enable the patterning of extruded bioinks in three-dimensional space. The support baths also present controllable mechanical cues for tuning the cytoskeletal properties of the patterned cells.


  • Publication

    Alexeev D, Cui S, Grad S, Li Z, Ferguson S. Mechanical and biological characterisation of a composite annulus fibrosus repair strategy in an endplate delamination model. JOR Spine. 2020 Jul 16;3(4):e1107. doi: 10.1002/jsp2.1107. eCollection 2020 Dec.

    Pfannkuche JJ, Guo W, Cui S, Ma J, Lang G, Peroglio M, Richards RG, Alini M, Grad S, Li Z. Intervertebral disc organ culture for the investigation of disc pathology and regeneration - benefits, limitations, and future directions of bioreactors. Connect Tissue Res. 61(3-4):304-321, 2020. doi: 10.1080/03008207.2019.1665652.

    Vernengo AJ, Grad S, Eglin D, Alin M, Li Z. Bioprinting Tissue Analogues with Decellularized Extracellular Matrix Bioink for Regeneration and Tissue Models of Cartilage and Intervertebral Discs. Adv. Funct. Mater. 2020, 1909044. DOI: 10.1002/adfm.201909044

    Kluser N. 3D Printed Multi-Scale Scaffolds with Topographical Guidance for Annulus Fibrosus Regeneration. 2020 ETH Zürich (A. Vernengo, S. Grad, S. Ferguson) – MSc ETH HST (Thesis)

    Zhang F, Liu X, Li B, Li Z, Grad S, Chen D, Gao M, Liu S.  The effect of hyaluronic acid on nucleus pulposus extracellular matrix production through hypoxia-inducible factor-1alpha transcriptional activation of CD44 under hypoxia.  Eur Cell Mater. 2021;41:142-52 https://www.doi.org/10.22203/eCM.v041a10

    Chen J-X, Li Y-H, Wen J, Li Z, Yu B-S, Huang Y-C. Annular defects impair the mechanical stability of the intervertebral disc. Global Spine J. 2021;epub Mar 30. https://doi.org/10.1177/21925682211006061

    Guo W, Douma L, Hu MH, Eglin D, Alini M, Secerovic A, Grad S, Peng X, Zou X, D'Este M, Peroglio M. Hyaluronic acid-based interpenetrating network hydrogel as a cell carrier for nucleus pulposus repair. Carbohydr Polym. 2021;277:118828. https://doi.org/10.1016/j.carbpol.2021.118828 

    Zhou L, Guo P, D'Este M, Tong W, Xu J, Yao H, Stoddart MJ, van Osch GJVM, Ki-Wai Ho K, Li Z, Qin L. Functionalized hydrogels for articular cartilage tissue engineering. Engineering. 2022;epub April 12, https://doi.org/10.1016/j.eng.2022.03.008

    Zhou J, Ren R, Li Z, Zhu S, Jiang N. Temporomandibular joint osteoarthritis: A review of animal models induced by surgical interventions. Oral Dis. 2022;epub May 25.  https://doi.org/10.1111/odi.14266 

    Russo F, Ambrosio L, Peroglio M, Guo W, Wangler S, Gewiess J, Grad S, Alini M, Papalia R, Vadalà G, Denaro V. A hyaluronan and platelet-rich plasma hydrogel for mesenchymal stem cell delivery in the intervertebral disc: an organ culture study. Int. J. Mol. Sci. 2021, 22, 2963.

    Guo W, Douma L, Hu MH, Eglin D, Alini M, Šećerović A, Grad S, Peng X, Zou X, D'Este M, Peroglio M. Hyaluronic acid-based interpenetrating network hydrogel as a cell carrier for nucleus pulposus repair. Carbohydr Polym. 2022 Feb 1;277:118828. DOI: 10.1016/j.carbpol.2021.118828.

    Lee NN, Salzer E, Bach FC, Bonilla AF, Cook JL, Gazit Z, Grad S, Ito K, Smith LJ, Vernengo A,Wilke HJ, Engiles JB, Tryfonidou MA. A comprehensive tool box for large animal studies of intervertebral disc degeneration. JOR Spine. 2021;4(2):e1162. https://doi.org/10.1002/jsp2.1162

     

     

     
  • Presentation

    Alig G, D'Este M, Grad S, Alini M, Vernengo A. Temperature-responsive engineered matrices for the spatial positioning and long-term culture of bioprinted cells.  2021 BIOFAB online (poster)

    Guo P, Vernengo A, Miklosic G, D'Este M, Grad S, Alini M, Li Z. A natural biomimetic hydrogel for cartilage regeneration. 2021 SSB+RM YSS (oral)

    Guo P, Mini C, Miklosic G, Vernengo A, D'Este M, Grad S, Alini M, Li Z. A hydrogel based on decellularized extracellular matrix particles for cartilage regeneration.
    2021 eCM cancelled (oral)/abstract published. https://www.ecmconferences.org/abstracts/2021/Collection3/oral.pdf 

    Egger S, Alig G, D'Este M, Grad S, Alini M, Vernengo A. Temperature-responsive engineered matrices for spatial positioning and long-term culture of bioprinted cells. 2021 eCM cancelled (poster)/abstract published. https://www.ecmconferences.org/abstracts/2021/Collection3/poster.pdf

    Egger S, Alig G, D'Este M, Wychowaniec JK, Weiser JR, Grad S, Vernengo AJ. Microextrusion-based anisotropic cell patterning within temperature-sensitive hydrogel matrices for annulus fibrosus regeneration. 2022 eCM (oral) 

    Egger S, Alig G, D'Este M, Grad S, Alini M, Vernengo A. 2021. 'Temperature-responsive engineered matrices for spatial positioning and long-term culture of bioprinted cells'. eCM XX: Biofabrication for Orthopaedics, Davos, Switzerland, (poster). 
     
    Alig G, D'Este M, Grad S, Alini M, Vernengo A. 2021. Temperature-responsive engineered hydrogel support baths for spatial positioning and long-term culture of bioprinted cells. International Conference on Biofabrication: Biofabrication 2021, virtual meeting, (paper and poster).
     
     

     

     
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