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J Reconstr Microsurg 2021; 37(04): 357-364
DOI: 10.1055/s-0040-1716855
Original Article

Mechanism of Peripheral Nerve Regeneration Using a Bio 3D Conduit Derived from Normal Human Dermal Fibroblasts

Hirofumi Yurie
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
,
Ryosuke Ikeguchi
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
,
Tomoki Aoyama
2   Department of Physical Therapy, Human Health Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan
,
Akira Ito
2   Department of Physical Therapy, Human Health Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan
,
Mai Tanaka
2   Department of Physical Therapy, Human Health Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan
,
Takashi Noguchi
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
,
Hiroki Oda
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
,
Hisataka Takeuchi
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
,
Sadaki Mitsuzawa
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
,
Maki Ando
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
,
Koichi Yoshimoto
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
,
Shizuka Akieda
3   Cyfuse Biomedical K.K., Tokyo, Japan
,
Koichi Nakayama
4   Department of Regenerative Medicine and Biomedical Engineering Faculty of Medicine, Saga University, Saga, Japan
,
Shuichi Matsuda
1   Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan
› Author Affiliations Funding This study was supported by JSPS KAKENHI, grant number: 15K10441. Cyfuse Biomedical K.K. provided the bioprinter “Regenova,” which was used in this experiment to construct the Bio three-dimensional conduit and contributed financially to this study under the Collaborative Research Agreement of Kyoto University. Cyfuse was not involved in any experimental methods, data collection, evaluation of the results, or preparation of the manuscript.
› Further Information
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Abstract

Background We previously reported the development of a scaffold-free Bio three-dimensional (3D) nerve conduit from normal human dermal fibroblasts (NHDFs). The aim of this study was to investigate the regenerative mechanism of peripheral nerve cells using a Bio 3D conduit in a rat sciatic nerve defect model.

Methods Bio 3D conduits composed of NHDFs were developed, and cell viability was evaluated using a LIVE/DEAD cell viability assay immediately before transplantation and 1-week post-surgery. Tracking analysis using PKH26-labeled NHDFs was performed to assess the distribution of NHDFs within the regenerated nerve and the differentiation of NHDFs into functional Schwann cells (SCs).

Results The assessment of the viability of cells within the Bio 3D conduit showed high cell viability both immediately before transplantation and 1-week post-surgery (88.56 ± 1.70 and 87.58 ± 9.11, respectively). A modified Masson's trichrome staining of the Bio 3D conduit revealed the formation of a prominent extracellular matrix (ECM) in between the cells. We observed, via tracking analysis, that the tube-like distribution of the NHDFs remained stable, the majority of the regenerated axons had penetrated this structure and PKH26-labeled cells were also positive for S-100.

Conclusion Abundant ECM formation resulted in a stable tube-like structure of the Bio 3D conduit with high cell viability. NHDFs in the Bio 3D conduit have the potential to differentiate into SCs-like cells.

Keywords

peripheral nerve - regeneration - bio 3D printer - fibroblast - angiogenesis


Publication History

Received: 06 June 2020

Accepted: 15 August 2020

Article published online:
21 September 2020

© 2020. Thieme. All rights reserved.

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