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J Reconstr Microsurg
DOI: 10.1055/a-2671-7768
Original Article

Adipose-Derived Stem Cell Sheets Prepared with Ascorbate 2-Phosphate Enhance Nerve Regeneration in Rat Sciatic Nerve Autografts

Atsuro Murai
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Kaoru Tada
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Mika Akahane
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Yuta Nakamura
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Soichiro Honda
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Masashi Matsuta
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Akari Mori
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Satoru Demura
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Jureepon Roboon
2   Department of Anatomy, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand
3   Centre of Excellence in Medical Biotechnology, Naresuan University, Phitsanulok, Thailand
,
Tsuyoshi Hattori
4   Department of Neuroanatomy, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Osamu Hori
4   Department of Neuroanatomy, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
,
Hiroyuki Tsuchiya
1   Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
5   Department of Orthopaedic Surgery, Yokohama Sakae Kyosai Hospital, Kanagawa, Japan
› Author Affiliations
Funding The authors would like to thank Applied Medical Research Laboratory for the preparation of sections with toluidine blue staining, and the Biopathology Institute Co., Ltd. for the pathology sections with DiI staining and pathology photographs. They would also like to thank Editage for English translation services. This work was supported by JSPS KAKENHI (grant number: JP20K22974).
› Further Information
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Abstract

Background

Autologous nerve grafts remain the gold standard for peripheral nerve repair, but have limited regenerative potential. Adipose-derived stem cells (ADSCs) have been investigated for their potential in nerve regeneration, and ascorbate 2-phosphate (A2P) enables the formation of ADSC sheets. This study examined whether ADSC sheets applied around autologous nerve grafts enhance functional and histological recovery in a rat sciatic nerve model.

Methods

A 15 mm sciatic nerve segment was excised, inverted, and sutured for autologous grafting in rats. Three groups were compared: phosphate-buffered saline (control), ADSC suspension, and ADSC sheets. Functional recovery was assessed at 12 weeks (n = 10 per group) using the sciatic functional index (SFI), nerve conduction studies (NCS; latency and amplitude), and tibialis anterior muscle wet weight. Histological analyses, including toluidine blue staining, evaluated axonal changes at 1, 2, 4, 8, and 12 weeks (n = 3 per group per time point). DiI-labeled ADSCs were tracked at 1 week to assess cell retention (n = 3 per group).

Results

At 12 weeks, the ADSC sheet group showed significantly improved SFI and muscle wet weight compared with controls and ADSC suspension groups. NCS revealed shorter distal latency in the ADSC sheet group versus controls, with no significant differences in the suspension group. While histological analysis did not demonstrate statistically significant differences among the groups, qualitative observations suggested that the ADSC sheet group tended to exhibit a greater number of myelinated axons at 12 weeks and fewer degenerative changes at earlier time points (1 and 2 weeks). DiI-labeled ADSCs were more frequently observed around the graft in the sheet group compared with the suspension group.

Conclusion

Application of ADSC sheets to autologous nerve grafts may promote functional recovery. Forming ADSC sheets with A2P may represent a favorable approach for improving outcomes in peripheral nerve repair.

Keywords

sciatic nerve - peripheral nerve regeneration - adipose-derived stem cells - autograft - Wallerian degeneration - cell sheet technology


Publication History

Received: 22 March 2025

Accepted: 27 July 2025

Accepted Manuscript online:
31 July 2025

Article published online:
21 August 2025

© 2025. Thieme. All rights reserved.

Thieme Medical Publishers, Inc.
333 Seventh Avenue, 18th Floor, New York, NY 10001, USA

 

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