Skip to main content

Advertisement

SpringerLink
Log in

Elastic Fiber-Reinforced Silk Fibroin Scaffold with A Double-Crosslinking Network for Human Ear-Shaped Cartilage Regeneration

  • Research Article
  • Published:
Advanced Fiber Materials Aims and scope Submit manuscript

Abstract

Tissue engineering provides a promising approach for regenerative medicine. The ideal engineered tissue should have the desired structure and functional properties suitable for uniform cell distribution and stable shape fidelity in the full period of in vitro culture and in vivo implantation. However, due to insufficient cell infiltration and inadequate mechanical properties, engineered tissue made from porous scaffolds may have an inconsistent cellular composition and a poor shape retainability, which seriously hinders their further clinical application. In this study, silk fibroin was integrated with silk short fibers with a physical and chemical double-crosslinking network to fabricate fiber-reinforced silk fibroin super elastic absorbent sponges (Fr-SF-SEAs). The Fr-SF-SEAs exhibited the desirable synergistic properties of a honeycomb structure, hygroscopicity and elasticity, which allowed them to undergo an unconventional cyclic compression inoculation method to significantly promote cell diffusion and achieve a uniform cell distribution at a high-density. Furthermore, the regenerated cartilage of the Fr-SF-SEAs scaffold withstood a dynamic pressure environment after subcutaneous implantation and maintained its precise original structure, ultimately achieving human-scale ear-shaped cartilage regeneration. Importantly, the SF-SEAs preparation showed valuable universality in combining chemicals with other bioactive materials or drugs with reactive groups to construct microenvironment bionic scaffolds. The established novel cell inoculation method is highly versatile and can be readily applied to various cells. Based on the design concept of dual-network Fr-SF-SEAs scaffolds, homogenous and mature cartilage was successfully regenerated with precise and complicated shapes, which hopefully provides a platform strategy for tissue engineering for various cartilage defect repairs.

Graphical Abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data availability

The data and materials are available.

References

  1. Lumelsky N, O’Hayre M, Chander P, Shum L, Somerman MJ. Autotherapies: enhancing endogenous healing and regeneration. Trends Mol Med. 2018;24:919.

    Article  Google Scholar 

  2. Wei W, Dai H. Articular cartilage and osteochondral tissue engineering techniques: recent advances and challenges. Bioact Mater. 2021;6:4830.

    Article  CAS  Google Scholar 

  3. Reid JA, Dwyer KD, Schmitt PR, Soepriatna AH, Coulombe KL, Callanan A. Architected fibrous scaffolds for engineering anisotropic tissues. Biofabrication. 2021, 13.

  4. Ju XJ, Liu XZ, Zhang Y, Chen X, Chen MM, Shen HS, Feng YH, Liu JS, Wang M, Shi Q. A photo-crosslinked proteinogenic hydrogel enabling self-recruitment of endogenous TGF-β1 for cartilage regeneration. Smart Mater in Med. 2022;3:85.

    Article  Google Scholar 

  5. Zhang YC, Gao HC, Luo HT, Chen DF, Zhou ZY, Cao XD. High strength HA-PEG/NAGA-Gelma double network hydrogel for annulus fibrosus rupture repair. Smart Mater in Med. 2022;3:128.

    Article  Google Scholar 

  6. Jia LT, Hua YJ, Zeng JS, Liu WS, Wang D, Zhou GD, Liu X, Jiang HY. Bioprinting and regeneration of auricular cartilage using a bioactive bioink based on microporous photocrosslinkable acellular cartilage matrix. Bioact Mater. 2022;16:66.

    Article  CAS  Google Scholar 

  7. Li Z, Gunn J, Chen MH, Cooper A, Zhang M. On-site alginate gelation for enhanced cell proliferation and uniform distribution in porous scaffolds. J Biomed Mater Res A. 2008;86:552.

    Article  Google Scholar 

  8. Radisic M, Euloth M, Yang L, Langer R, Freed LE, Vunjak-Novakovic G. High-density seeding of myocyte cells for cardiac tissue engineering. Biotechnol Bioeng. 2003;82:403.

    Article  CAS  Google Scholar 

  9. Miller JS, Stevens KR, Yang MT, Baker BM, Nguyen DH, Cohen DM, Toro E, Chen AA, Galie PA, Yu X, Chaturvedi R, Bhatia SN, Chen CS. Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues. Nat Mater. 2012;11:768.

    Article  CAS  Google Scholar 

  10. Sari M, Hening P, Chotimah AID, Yusuf Y. Bioceramic hydroxyapatite-based scaffold with a porous structure using honeycomb as a natural polymeric Porogen for bone tissue engineering. Biomater Res. 2021;25:2.

    Article  CAS  Google Scholar 

  11. Buenzli PR, Lanaro M, Wong CS, McLaughlin MP, Allenby MC, Woodruff MA, Simpson MJ. Cell proliferation and migration explain pore bridging dynamics in 3D printed scaffolds of different pore size. Acta Biomater. 2020;114:285.

    Article  CAS  Google Scholar 

  12. Cengiz IF, Oliveira JM, Reis RL. Micro-CT—a digital 3D microstructural voyage into scaffolds: a systematic review of the reported methods and results. Biomater Res. 2018;22:26.

    Article  Google Scholar 

  13. Ci Z, Zhang Y, Wang Y, Wu GY, Hou MJ, Zhang PL, Jia LT, Bai BS, Cao YL, Liu Y, Zhou GD. 3D cartilage regeneration with certain shape and mechanical strength based on engineered cartilage gel and decalcified bone matrix. Front Cell Dev Biol. 2021;9: 638115.

    Article  Google Scholar 

  14. Lee M, Bae K, Guillon P, Chang J, Arlov Ø, Zenobi-Wong M. Exploitation of cationic silica nanoparticles for bioprinting of large-scale constructs with high printing fidelity. ACS Appl Mater Interfaces. 2018;10:37820.

    Article  CAS  Google Scholar 

  15. Cengiz IF, Pereira H, Espregueira-Mendes J, Oliveira JM, Reis RL. Treatments of meniscus lesions of the knee: current concepts and future perspectives. Regen Eng Transl Med. 2017;3:32.

    Article  Google Scholar 

  16. Wang Y, Kim HJ, Vunjak-Novakovic G, Kaplan DL. Stem cell-based tissue engineering with silk biomaterials. Biomaterials. 2006;27:6064.

    Article  CAS  Google Scholar 

  17. Omenetto FG, Kaplan DL. New opportunities for an ancient material. Science. 2010;329:528.

    Article  CAS  Google Scholar 

  18. Wang Y, Rudym DD, Walsh A, Abrahamsen L, Kim HJ, Kim HS, Kirker-Head C, Kaplan DL. In vivo degradation of three-dimensional silk fibroin scaffolds. Biomaterials. 2008;29:3415.

    Article  CAS  Google Scholar 

  19. Zhao CX, Liu YW, Lv ZC, Cao LT, Ren J, Shao ZZ, Ling SJ. Silk Fibroin Nacre. Adv Fiber Mater. 2022;4:1191.

    Article  CAS  Google Scholar 

  20. Shao ZZ, Vollrath F. Surprising strength of silkworm silk. Nature. 2002;418:741.

    Article  CAS  Google Scholar 

  21. Rousseau ME, Lefèvre T, Beaulieu L, Asakura T, Pézolet M. Study of protein conformation and orientation in silkworm and spider silk fibers using Raman microspectroscopy. Biomacromol. 2004;5:2247.

    Article  CAS  Google Scholar 

  22. Zhang Y, Lu HJ, Liang XP, Zhang MC, Liang HR, Zhang YY. Silk materials for intelligent fibers and textiles: potential, progress and future perspective. Acta Phys Chim Sin. 2022;38:2103034.

    Google Scholar 

  23. Lu HJ, Xia KL, Jian MQ, Liang XP, Yin Z, Zhang MC, Wang HM, Wang HM, Li S, Zhang YY. Mechanically reinforced silkworm silk fiber by hot stretching. Research (Wash D C). 2022;2022:9854063.

    CAS  Google Scholar 

  24. Zou SZ, Fan SN, Oliveira AL, Yao X, Zhang YP, Shao HL. 3D printed gelatin scaffold with improved shape fidelity and cytocompatibility by using antheraea pernyi silk fibroin nanofibers. Adv Fiber Mater. 2022;4:758.

    Article  CAS  Google Scholar 

  25. Lu HJ, Jian MQ, Yin Z, Xia KL, Shi SY, Zhang MC, Wang HM, Liang XP, Ma WG, Zhang X, Zhang YY. Silkworm silk fibers with multiple reinforced properties obtained through feeding Ag nanowires. Adv Fiber Mater. 2022;4:547.

    Article  CAS  Google Scholar 

  26. Wang Q, Ling SJ, Yao QZ, Li QY, Hu DB, Dai Q, Weitz DA, Kaplan DL, Buehler MJ, Zhang YY. Observations of 3 nm silk nanofibrils exfoliated from natural silkworm silk fibers. ACS Mater Lett. 2020;2:153.

    Article  CAS  Google Scholar 

  27. Zhang J, Allardyce BJ, Rajkhowa R, Zhao Y, Dilley RJ, Redmond SL, Wang X, Liu X. 3D printing of silk particle-reinforced chitosan hydrogel structures and their properties. ACS Biomater Sci Eng. 2018;4:3036.

    Article  CAS  Google Scholar 

  28. Huang L, Du XY, Fan SN, Yang GS, Shao HL, Li DJ, Cao CB, Zhu YF, Zhu MF, Zhang YP. Bacterial cellulose nanofibers promote stress and fidelity of 3D-printed silk based hydrogel scaffold with hierarchical pores. Carbohydr Polym. 2019;221:146.

    Article  CAS  Google Scholar 

  29. Mandal BB, Grinberg A, Gil ES, Panilaitis B, Kaplan DL. High-strength silk protein scaffolds for bone repair. Proc Natl Acad Sci USA. 2012;109:7699.

    Article  CAS  Google Scholar 

  30. Jia LT, Zhang Y, Yao L, Zhang PL, Ci Z, Zhang W, Miao CL, Liang XQ, He AJ, Liu Y, Tang SJ, Zhang RH, Wang XY, Cao YL, Zhou GD. Regeneration of human-ear-shaped cartilage with acellular cartilage matrix-based biomimetic scaffolds. Appl Mater Today. 2020;20: 100639.

    Article  Google Scholar 

  31. Lei D, Yang Y, Liu ZH, Yang BQ, Gong WH, Chen S, Wang SF, Sun LJ, Song BY, Xuan HX, Mo XM, Sun BB, Li S, Yang Q, Huang SX, Chen SY, Ma YD, Liu WG, He CL, Zhu B, Jeffries EM, Qing FL, Ye XF, Zhao Q, You ZW. 3D printing of biomimetic vasculature for tissue regeneration. Mater Horiz. 2019;6:1197.

    Article  CAS  Google Scholar 

  32. Rodriguez A, Cao YL, Ibarra C, Pap S, Vacanti M, Eavey RD, Vacanti CA. Characteristics of cartilage engineered from human pediatric auricular cartilage. Plast Reconstr Surg. 1999;103:1111.

    Article  CAS  Google Scholar 

  33. Liu Y, Zhang L, Zhou GD, Li Q, Liu W, Yu ZY, Luo XS, Jiang T, Zhang WJ, Cao YL. In vitro engineering of human ear-shaped cartilage assisted with CAD/CAM technology. Biomaterials. 2010;31:2176.

    Article  CAS  Google Scholar 

  34. Zhou GD, Jiang HY, Yin ZQ, Liu Y, Zhang QG, Zhang C, Pan B, Zhou JY, Zhou X, Sun HY, Li D, He AJ, Zhang ZY, Zhang WJ, Liu W, Cao YL. In vitro regeneration of patient-specific ear-shaped cartilage and its first clinical application for auricular reconstruction. EBioMedicine. 2018;28:287.

    Article  Google Scholar 

  35. Li Q, Liu TY, Zhang L, Liu Y, Zhang WJ, Liu W, Cao YL, Zhou GD. The role of bFGF in down-regulating α-SMA expression of chondrogenically induced BMSCs and preventing the shrinkage of BMSC engineered cartilage. Biomaterials. 2011;32:4773.

    Article  CAS  Google Scholar 

  36. He AJ, Xia HT, Xiao KY, Wang TT, Liu Y, Xue JX, Li D, Tang SJ, Liu FJ, Wang XY, Zhang WJ, Liu W, Cao YL, Zhou GD. Cell yield, chondrogenic potential, and regenerated cartilage type of chondrocytes derived from ear, nasoseptal, and costal cartilage. J Tissue Eng Regen Med. 2018;12:1123.

    Article  CAS  Google Scholar 

  37. Li D, Yin ZQ, Liu Y, Feng SQ, Liu Y, Lu FJ, Xu Y, Min PR, Hou MJ, Li K, He AJ, Zhang WJ, Liu W, Zhang YX, Zhou GD, Cao YL. Regeneration of trachea graft with cartilage support, vascularization, and epithelization. Acta Biomater. 2019;89:206.

    Article  CAS  Google Scholar 

  38. Luo XS, Zhou GD, Liu W, Zhang WJ, Cen L, Cui L, Cao YL. In vitro precultivation alleviates post-implantation inflammation and enhances development of tissue-engineered tubular cartilage. Biomed Mater. 2009;4: 025006.

    Article  Google Scholar 

  39. Falsafi S, Koch RJ. Growth of tissue-engineered human nasoseptal cartilage in simulated microgravity. Arch Otolaryngol Head Neck Surg. 2000;126:759.

    Article  CAS  Google Scholar 

  40. Yin ZQ, Li D, Liu Y, Feng SQ, Yao L, Liang XQ, Miao CL, Xu Y, Hou MJ, Zhang RH, Zhang WJ, Liu W, Liu Y, Zhou GD, Cao YL. Regeneration of elastic cartilage with accurate human-ear shape based on PCL strengthened biodegradable scaffold and expanded microtia chondrocytes. Appl Mater Today. 2020;20: 100724.

    Article  Google Scholar 

  41. Visser J, Melchels FP, Jeon JE, van Bussel EM, Kimpton LS, Byrne HM, Dhert WJ, Dalton PD, Hutmacher DW, Malda J. Reinforcement of hydrogels using three-dimensionally printed microfibres. Nat Commun. 2015;6:6933.

    Article  CAS  Google Scholar 

  42. Mohabatpour F, Karkhaneh A, Sharifi AM. A hydrogel/fiber composite scaffold for chondrocyte encapsulation in cartilage tissue regeneration. RSC Adv. 2016;6:83135.

    Article  CAS  Google Scholar 

  43. Shen YB, Xu Y, Yi BC, Wang XL, Tang H, Chen C, Zhang YZ. Engineering a highly biomimetic chitosan-based cartilage scaffold by using short fibers and a cartilage-decellularized matrix. Biomacromol. 2021;22:2284.

    Article  CAS  Google Scholar 

  44. Muncie JM, Weaver VM. The physical and biochemical properties of the extracellular matrix regulate cell fate. Curr Top Dev Biol. 2018;130:1.

    Article  CAS  Google Scholar 

  45. Soans KG, Norden C. Shining a light on extracellular matrix dynamics in vivo. Semin Cell Dev Biol. 2021;120:85.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We gratefully acknowledge financial support from the National Key Research and Development Program of China (2018YFC1105800, 2017YFC1103900), the National Natural Science Foundation of China (82102211, 81871502), the Shanghai Municipal Key Clinical Specialty (shslczdzk06601), the Shanghai Collaborative Innovation Program on Regenerative Medicine and Stem Cell Research (2019CXJQ01), the Key Research and Development Program of Henan Province (No. 221111310100), the Major Science and Technology Projects of Xinxiang City (No. 21ZD006), the Clinical Research Plan of SHDC (No. SHDC2020CR2045B), and the Start-up Funds of Talent Construction and Scientific Research in Shanghai 9th People's Hospital (2021rcyj-ld).

Author information

Author notes

  1. Qianyi Wang, Xinyue Ran and Jian Wang have contributed equally to the work.

Authors and Affiliations

  1. Department of Plastic and Reconstructive Surgery, Shanghai 9th People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Lab of Tissue Engineering, Shanghai, 200011, People’s Republic of China

    Qianyi Wang, Xinyue Ran, Jian Wang, Sinan Wang, Peiling Zhang, Erji Gao, Baoshuai Bai, Junfeng Zhang, Guangdong Zhou & Dong Lei

  2. Research Institute of Plastic Surgery, Weifang Medical University, Weifang, 261053, People’s Republic of China

    Qianyi Wang, Xinyue Ran, Sinan Wang & Guangdong Zhou

  3. Department of Cardiology, Shanghai 9th People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, People’s Republic of China

    Junfeng Zhang & Dong Lei

Authors
  1. Qianyi Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xinyue Ran
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jian Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sinan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peiling Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Erji Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Baoshuai Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Junfeng Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Guangdong Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Dong Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Junfeng Zhang, Guangdong Zhou or Dong Lei.

Ethics declarations

Conflicts of interest

The authors declare no competing financial interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 599 KB)

Supplementary file2 (MP4 2859 KB)

Supplementary file3 (MP4 1398 KB)

Supplementary file4 (MP4 851 KB)

Supplementary file5 (MP4 724 KB)

Supplementary file6 (MP4 3295 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, Q., Ran, X., Wang, J. et al. Elastic Fiber-Reinforced Silk Fibroin Scaffold with A Double-Crosslinking Network for Human Ear-Shaped Cartilage Regeneration. Adv. Fiber Mater. 5, 1008–1024 (2023). https://doi.org/10.1007/s42765-023-00266-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s42765-023-00266-8

Keywords

Search

Navigation