AccScience Publishing / IJB / Volume 10 / Issue 1 / DOI: 10.36922/ijb.0161
Submit to IJB
Cite this article
221
Download
1446
Views
Journal Browser
Volume | Year
Issue
Search
Forthcoming Issue
Current Issue
View All
News and Announcements
View All
RESEARCH ARTICLE

Chondrocyte spheroid-laden microporous hydrogel-based 3D bioprinting for cartilage regeneration

Ruiquan Liu1 ,  Litao Jia2 ,  Jianguo Chen1 ,  Yi Long3 ,  Jinshi Zeng1 ,  Siyu Liu2 ,  Bo Pan1 ,  Xia Liu2* ,  Haiyue Jiang1*
Show Less
1 Department of Auricular Reconstruction, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100144, China
2 Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100144, China
3 Department of Biomedical Engineering, School of Medicine, Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing 10084, China
IJB 2024, 10(1), 0161 https://doi.org/10.36922/ijb.0161
Submitted: 11 May 2023 | Accepted: 14 June 2023 | Published: 28 July 2023
© 2023 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Download PDF
Cite
XML
Abstract

Three-dimensional (3D) bioprinting has brought new promising strategies for the regeneration of cartilage with specific shapes. In cartilage bioprinting, chondrocyte-laden hydrogels are the most commonly used bioinks. However, the dispersion of cells and the dense texture of the hydrogel in the conventional bioink may limit cell–cell/ cell–extracellular matrix (ECM) interactions, counting against cartilage regeneration and maturation. To address this issue, in this study, we developed a functional bioink for cartilage bioprinting based on chondrocyte spheroids (CSs) and microporous hydrogels, in which CSs as multicellular aggregates can provide extensive cell– cell/cell–ECM interactions to mimic the natural cartilage microenvironment, and microporous hydrogels can provide space and channel for the growth and fusion of the CSs. Firstly, we used a non-adhesive microporous system to produce homogeneous self-assembled CSs in high-throughput and evaluated the influence of different CSs preparation parameters (cell number and culture time) on CSs, which aids in the preparation of bioink suitable for cartilage bioprinting. Then, polyethylene oxide (PEO) was introduced into gelatin methacrylate (GelMA) to prepare microporous hydrogel. Finally, the CS-laden microporous hydrogels were printed, and the constructs were implanted into nude mice. The results showed that the CSs with 500 cells cultured for 1 day exhibited better proliferation and growth ability in microporous hydrogels compared to those with more cells and cultured for longer time. In addition, the results also demonstrated that the CS-laden bioink can be successfully printed into predefined lattice-shape constructs with little cell damage and regenerated cartilage tissue in vivo with a structure similar to natural cartilage characterized by typical lacunae structure and abundant cartilage-specific ECM deposition. In summary, our study verified the feasibility and advantages of using CSs as building blocks in cartilage bioprinting, which provides novel strategies for the fabrication and regeneration of patient-specific shaped cartilage.

Keywords
3D bioprinting
Chondrocyte spheroids
Microporous hydrogels
Cartilage regeneration
Funding
This work was supported by the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences (2021-I2M-1-052), the National Natural Science Foundation of China (81871575), and the Fundamental Research Funds for the Central Universities (3332022139).
References
  1. Jessop ZM, Hague A, Dobbs TD, Stewart KJ, Whitaker IS. Facial cartilaginous reconstruction-A historical perspective, state-of-the-art, and future directions. Front Surg. 2021;8:680186. doi: 10.3389/fsurg.2021.680186
  2. Cao Y, Sang S, An Y, Xiang C, Li Y, Zhen Y. Progress of 3D printing techniques for nasal cartilage regeneration. Aesthetic Plast Surg. 2022;46(2):947–964. doi: 10.1007/s00266-021-02472-4
  3. Cao Y, Vacanti JP, Paige KT, Upton J, Vacanti CA. Transplantation of chondrocytes utilizing a polymer-cell construct to produce tissue-engineered cartilage in the shape of a human ear. Plast Reconstr Surg. 1997,100(2): 297–302; discussion 303–294. doi: 10.1097/00006534-199708000-00001
  4. Tan B, Gan S, Wang X, Liu W, Li Xiaoming. Applications of 3D bioprinting in tissue engineering: Advantages, deficiencies, improvements, and future perspectives. J Mater Chem B. 2021;9(27):5385–5413. doi: 10.1039/d1tb00172h
  5. Wang G, Zhang X, Bu X, Yang An, Bi Hongsen, Zhao Z. The application of cartilage tissue engineering with cell-laden hydrogel in plastic surgery: A systematic review. Tissue Eng Regen Med. 2022;19(1):1–9. doi: 10.1007/s13770-021-00394-5
  6. Tang P, Song P, Peng Z, et al. Chondrocyte-laden GelMA hydrogel combined with 3D printed PLA scaffolds for auricle regeneration. Mater Sci Eng C Mater Biol Appl. 2021;130(1):112423. doi: 10.1016/j.msec.2021.112423
  7. Zeng J, Jia L, Wang D, et al. Bacterial nanocellulose-reinforced gelatin methacryloyl hydrogel enhances biomechanical property and glycosaminoglycan content of 3D-bioprinted cartilage. Int J Bioprint. 2023;9(1):631. doi: 10.18063/ijb.v9i1.631
  8. Achilli TM, Meyer J, Morgan JR. Advances in the formation, use and understanding of multi-cellular spheroids. Expert Opin Biol Ther. 2012;12(10):1347–1360. doi: 10.1517/14712598.2012.707181
  9. Kronemberger GS, Matsui RAM, Miranda G, Granjeiro JM, Baptista LS. Cartilage and bone tissue engineering using adipose stromal/stem cells spheroids as building blocks. World J Stem Cells. 2020;12(2):110–122. doi: 10.4252/wjsc.v12.i2.110
  10. Zhang J, Xin W, Qin Y, et al. “All-in-one” zwitterionic granular hydrogel bioink for stem cell spheroids production and 3D bioprinting. Chem Eng J. 2022;430(50):132713. doi: 10.1016/j.cej.2021.132713
  11. Chen H, Tan XN, Hu S, et al. Molecular mechanisms of chondrocyte proliferation and differentiation. Front Cell Dev Biol. 2021;9:664168.doi: 10.3389/fcell.2021.664168
  12. Ghosh S, Laha M, Mondal S, Sengupta S, Kaplana DL. In vitro model of mesenchymal condensation during chondrogenic development. Biomaterials. 2009;30(33): 6530–6540. doi: 10.1016/j.biomaterials.2009.08.019
  13. Chae S, Hong J, Hwangbo H, Kim G. The utility of biomedical scaffolds laden with spheroids in various tissue engineering applications. Theranostics. 2021;11(14): 6818–6832. doi: 10.7150/thno.58421
  14. Decarli MC, Amaral R, Santos DPD, et al. Cell spheroids as a versatile research platform: formation mechanisms, high throughput production, characterization and applications. Biofabrication. 2021;13(3):1-37. doi: 10.1088/1758-5090/abe6f2
  15. Mironov V, Visconti RP, Kasyanov V, Gabor Forgacs, Drake CJ, Markwald RR. Organ printing: Tissue spheroids as building blocks. Biomaterials. 2009;30(12):2164–2174. doi: 10.1016/j.biomaterials.2008.12.084
  16. Grevenstein D, Mamilos A, Schmitt VH, et al. Excellent histological results in terms of articular cartilage regeneration after spheroid-based autologous chondrocyte implantation (ACI). Knee Surg Sports Traumatol Arthrosc. 2021;29(2):417–421. doi: 10.1007/s00167-020-05976-9
  17. Gryadunova A, Kasamkattil J, Gay MHP, et al. Nose to spine: Spheroids generated by human nasal chondrocytes for scaffold-free nucleus pulposus augmentation. Acta Biomater. 2021;134(4):240–251. doi: 10.1016/j.actbio.2021.07.064
  18. Huang BJ, Hu JC, Athanasiou KA. Effects of passage number and post-expansion aggregate culture on tissue engineered, self-assembled neocartilage. Acta Biomater. 2016;43(3): 150–159. doi: 10.1016/j.actbio.2016.07.044
  19. Jeon JH, Yun BG, Lim MJ, et al. Rapid cartilage regeneration of spheroids composed of human nasal septum-derived chondrocyte in rat osteochondral defect model. Tissue Eng Regen Med. 2020;17(1):81–90. doi: 10.1007/s13770-019-00231-w
  20. Wang G, An Y, Zhang X, Pengbing Ding, Bi H, Zhao Z. Chondrocyte spheroids laden in GelMA/HAMA hybrid hydrogel for tissue-engineered cartilage with enhanced proliferation, better phenotype maintenance, and natural morphological structure. Gels. 2021;7(4):247. doi: 10.3390/gels7040247
  21. De Moor L, Minne M, Tytgat L, et al. Tuning the phenotype of cartilage tissue mimics by varying spheroid maturation and methacrylamide-modified gelatin hydrogel characteristics. Macromol Biosci. 2021;21(5):e2000401. doi: 10.1002/mabi.202000401
  22. Kang HW, Lee SJ, Ko IK, Yoo JJ, Atala A. A 3D bioprinting system to produce human-scale tissue constructs with structural integrity. Nat Biotechnol. 2016;34(3):312–319. doi: 10.1038/nbt.3413
  23. Ying GL, Jiang N, Maharjan S, et al. Aqueous two-phase emulsion bioink-enabled 3D bioprinting of porous hydrogels. Adv Mater. 2018;30(50):e1805460. doi: 10.1002/adma.201805460
  24. Ying G, Jiang N, Parra C, et al. Bioprinted injectable hierarchically porous gelatin methacryloyl hydrogel constructs with shape-memory properties. Adv Funct Mater. 2020;30(46). doi: 10.1002/adfm.202003740
  25. Shi W, Shi H, Fu G, et al. Application of agarose gelatin in tissue pre-embedding before dehydration. J Clin Exp Pathol. 2021;37(3):365–366. doi: 10.13315/j.cnki.cjcep.2021.03.031
  26. Jia L, Hua Y, Zeng J, et al. Bioprinting and regeneration of auricular cartilage using a bioactive bioink based on microporous photocrosslinkable acellular cartilage matrix. Bioact Mater. 2022;16(1):66–81. doi: 10.1016/j.bioactmat.2022.02.032
  27. Van Winkle AP, Gates ID, Kallos MS. Mass transfer limitations in embryoid bodies during human embryonic stem cell differentiation. Cells Tissues Organs. 2012;196(1):34–47. doi: 10.1159/000330691
  28. Lin RZ, Chang HY. Recent advances in three-dimensional multicellular spheroid culture for biomedical research. Biotechnol J. 2008;3(9–10):1172–1184. doi: 10.1002/biot.200700228
  29. Shi W, Kwon J, Huang Y, et al. Facile tumor spheroids formation in large quantity with controllable size and high uniformity. Sci Rep. 2018;8(1):6837. doi: 10.1038/s41598-018-25203-3
  30. Niebruegge S, Bauwens CL, Peerani R, et al. Generation of human embryonic stem cell-derived mesoderm and cardiac cells using size-specified aggregates in an oxygen-controlled bioreactor. Biotechnol Bioeng. 2009;102(2):493–507. doi: 10.1002/bit.22065
  31. Sart S, Tsai AC, Li Y, et al. Three-dimensional aggregates of mesenchymal stem cells: Cellular mechanisms, biological properties, and applications. Tissue Eng Part B Rev. 2014;20(5):365–380. doi: 10.1089/ten.TEB.2013.0537
  32. Behan K, Dufour A, Garcia O, Kelly D. Methacrylated cartilage ECM-based hydrogels as injectables and bioinks for cartilage tissue engineering. Biomolecules. 2022;12(2):216. doi: 10.3390/biom12020216
  33. 33. Ozbolat IT, Hospodiuk M. Current advances and future perspectives in extrusion-based bioprinting. Biomaterials. 2016;76:321–343. doi: 10.1016/j.biomaterials.2015.10.076
Conflict of interest
The authors declare no conflicts of interest.
Share
Back to top
International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing
We use cookies on our website to ensure you get the best experience.
Read more about our cookies here
Accept