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

Kaneda, Giselle<sup>1,2,*</sup>; Chan, Julie L.<sup>1,3,*</sup>; Castaneda, Chloe M.<sup>1,2</sup>; Papalamprou, Angela<sup>1,2</sup>; Sheyn, Julia<sup>1,2</sup>; Shelest, Oksana<sup>2</sup>; Huang, Dave<sup>4,5</sup>; Kluser, Nadine<sup>6</sup>; Yu, Victoria<sup>1,2</sup>; Ignacio, Gian C.<sup>4,5</sup>; Gertych, Arkadiusz<sup>7,8</sup>; Yoshida, Ryu<sup>5</sup>; Metzger, Melodie F.<sup>4,5</sup>; Tawackoli, Wafa<sup>1,2,5,7,9</sup>; Vernengo, Andrea<sup>6</sup>; Sheyn, Dmitriy<sup>1,2,5,7,9</sup>

Affiliations

¹Orthopaedic Stem Cell Research Laboratory, Cedars‐Sinai Medical Center, Los Angeles, California, USA

²Board of Governors Regenerative Medicine Institute, Cedars‐Sinai Medical Center, Los Angeles, California, USA

³Department of Neurosurgery, Cedars‐Sinai Medical Center, Los Angeles, California, USA

⁴Orthopedics Biomechanics Laboratory, Cedars‐Sinai Medical Center, Los Angeles, California, USA

⁵Department of Orthopedics, Cedars‐Sinai Medical Center, Los Angeles, California, USA

⁶AO Research Institute Davos, Davos, Switzerland

⁷Department of Surgery, Cedars‐Sinai Medical Center, Los Angeles, California, USA

⁸Department of Pathology and Laboratory Medicine, Cedars‐Sinai Medical Center, Los Angeles, California, USA

⁹Department of Biomedical Sciences, Cedars‐Sinai Medical Center, Los Angeles, California, USA

Abstract and keywords

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

Links

DOI: 10.1002/jor.25554

PubMed: 36961351

WoS: 000963133100001

History

Received: 2022-11-03

Revised: 2023-03-06

Accepted: 2023-0311