A translational approach integrating developmental biology and tissue engineering towards regeneration of the annulus fibrosus of the intervertebral disc (PrintDisc)

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.

Summary of the 3D bioprinting system for annulus fibrosus (AF) tissue engineering.

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

Zhou Z, Wang W, Wang J, Wang H, Wang W, Lai Y, Lin X, Huang Y, Zou X, Stoddart M, Li Z, Tian W, Liu S, Wu X, Gao M, Li J, Yang L, Chen D. Function-oriented design: A novel strategy for advanced biomedical materials. J Mat Sci Technol. 2023;145:197-209. https://doi.org/10.1016/j.jmst.2022.09.066

Ristaniemi A, Šećerović A, Dischl V, Crivelli F, Heub S, Ledroit D, Weder G, Grad S, Ferguson SJ. Physiological and degenerative loading of bovine intervertebral disc in a bioreactor: A finite element study of complex motions. J Mech Behav Biomed Mater. 2023;143:105900. https://doi.org/10.1016/j.jmbbm.2023.105900
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).

Grad S. Simulation of physiological and detrimental loading in whole intervertebral disc organ models. 2023 EORS (oral)