iPSC-derived tenocytes seeded on microgrooved 3D printed scaffolds for Achilles tendon regeneration

Giselle Kaneda; Julie L Chan; Chloe M Castaneda; Angela Papalamprou; Julia Sheyn; Oksana Shelest; Dave Huang; Nadine Kluser; Victoria Yu; Gian C Ignacio; Arkadiusz Gertych; Ryu Yoshida; Melodie F Metzger; Wafa Tawackoli; Andrea Vernengo; Dmitriy Sheyn

doi:10.1002/jor.25554

iPSC-derived tenocytes seeded on microgrooved 3D printed scaffolds for Achilles tendon regeneration

J Orthop Res. 2023 Oct;41(10):2205-2220. doi: 10.1002/jor.25554. Epub 2023 Apr 5.

Authors

Giselle Kaneda^{1

2}, Julie L Chan^{1

3}, Chloe M Castaneda^{1

2}, Angela Papalamprou^{1

2}, Julia Sheyn^{1

2}, Oksana Shelest², Dave Huang^{4

5}, Nadine Kluser⁶, Victoria Yu^{1

2}, Gian C Ignacio^{4

5}, Arkadiusz Gertych^{7

8}, Ryu Yoshida⁵, Melodie F Metzger^{4

5}, Wafa Tawackoli^{1

2

5

7

9}, Andrea Vernengo⁶, Dmitriy Sheyn^{1

2

5

7

9}

Affiliations

¹ Orthopaedic Stem Cell Research Laboratory, Cedars-Sinai Medical Center, Los Angeles, California, USA.
² Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, California, USA.
³ Department of Neurosurgery, Cedars-Sinai Medical Center, Los Angeles, California, USA.
⁴ Orthopedics Biomechanics Laboratory, Cedars-Sinai Medical Center, Los Angeles, California, USA.
⁵ Department of Orthopedics, Cedars-Sinai Medical Center, Los Angeles, California, USA.
⁶ AO Research Institute Davos, Davos, Switzerland.
⁷ Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, California, USA.
⁸ Department of Pathology and Laboratory Medicine, Cedars-Sinai Medical Center, Los Angeles, California, USA.
⁹ Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.

PMID: 36961351
PMCID: PMC10518032 (available on 2024-10-01)
DOI: 10.1002/jor.25554

Abstract

Tendons and ligaments have a poor innate healing capacity, yet account for 50% of musculoskeletal injuries in the United States. Full structure and function restoration postinjury remains an unmet clinical need. This study aimed to assess the application of novel three dimensional (3D) printed scaffolds and induced pluripotent stem cell-derived mesenchymal stem cells (iMSCs) overexpressing the transcription factor Scleraxis (SCX, iMSC^SCX+ ) as a new strategy for tendon defect repair. The polycaprolactone (PCL) scaffolds were fabricated by extrusion through a patterned nozzle or conventional round nozzle. Scaffolds were seeded with iMSC^SCX+ and outcomes were assessed in vitro via gene expression analysis and immunofluorescence. In vivo, rat Achilles tendon defects were repaired with iMSC^SCX+ -seeded microgrooved scaffolds, microgrooved scaffolds only, or suture only and assessed via gait, gene expression, biomechanical testing, histology, and immunofluorescence. iMSC^SCX+ -seeded on microgrooved scaffolds showed upregulation of tendon markers and increased organization and linearity of cells compared to non-patterned scaffolds in vitro. In vivo gait analysis showed improvement in the Scaffold + iMSC^SCX+ -treated group compared to the controls. Tensile testing of the tendons demonstrated improved biomechanical properties of the Scaffold + iMSC^SCX+ group compared with the controls. Histology and immunofluorescence demonstrated more regular tissue formation in the Scaffold + iMSC^SCX+ group. This study demonstrates the potential of 3D-printed scaffolds with cell-instructive surface topography seeded with iMSC^SCX+ as an approach to tendon defect repair. Further studies of cell-scaffold constructs can potentially revolutionize tendon reconstruction by advancing the application of 3D printing-based technologies toward patient-specific therapies that improve healing and functional outcomes at both the cellular and tissue level.

Keywords: Achilles tendon; iPSC; microgrooves; scaffold; scleraxis.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Achilles Tendon*
Animals
Induced Pluripotent Stem Cells*
Printing, Three-Dimensional
Rats
Regeneration
Tenocytes
Tissue Engineering / methods
Tissue Scaffolds / chemistry
Wound Healing

Grants and funding

K01 AR071512/AR/NIAMS NIH HHS/United States