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

Hirofumi Yurie; Ryosuke Ikeguchi; Tomoki Aoyama; Akira Ito; Mai Tanaka; Takashi Noguchi; Hiroki Oda; Hisataka Takeuchi; Sadaki Mitsuzawa; Maki Ando; Koichi Yoshimoto; Shizuka Akieda; Koichi Nakayama; Shuichi Matsuda

doi:10.1055/s-0040-1716855

Journal of Reconstructive Microsurgery, Table of Contents

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

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

,

Ryosuke Ikeguchi

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

,

Tomoki Aoyama

²Department of Physical Therapy, Human Health Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan

,

Akira Ito

²Department of Physical Therapy, Human Health Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan

,

Mai Tanaka

²Department of Physical Therapy, Human Health Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan

,

Takashi Noguchi

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

,

Hiroki Oda

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

,

Hisataka Takeuchi

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

,

Sadaki Mitsuzawa

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

,

Maki Ando

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

,

Koichi Yoshimoto

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

,

Shizuka Akieda

³Cyfuse Biomedical K.K., Tokyo, Japan

,

Koichi Nakayama

⁴Department of Regenerative Medicine and Biomedical Engineering Faculty of Medicine, Saga University, Saga, Japan

,

Shuichi Matsuda

¹Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan

› Author Affiliations

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

Full Text

References

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