Comparison of ligamentization potential between anterior cruciate ligament-derived cells and adipose-derived mesenchymal stem cells reseeded to acellularized tendon allograft

Jinsung Park; Sungsin Jo; Myung-Kyu Lee; Tae-Hwan Kim; Il-Hoon Sung; Jin K Lee

doi:10.1302/2046-3758.1111.BJR-2021-0548.R2

Comparison of ligamentization potential between anterior cruciate ligament-derived cells and adipose-derived mesenchymal stem cells reseeded to acellularized tendon allograft

Bone Joint Res. 2022 Nov;11(11):777-786. doi: 10.1302/2046-3758.1111.BJR-2021-0548.R2.

Authors

Jinsung Park^{1

2}, Sungsin Jo¹, Myung-Kyu Lee³, Tae-Hwan Kim^{1

2

4}, Il-Hoon Sung⁵, Jin K Lee⁵

Affiliations

¹ Hanyang University Institute for Rheumatology Research (HYIRR), Seoul, South Korea.
² Department of Translational Medicine, Graduate School of Biomedical Science and Engineering, Hanyang University, Seoul, South Korea.
³ Department of Research and Development, Korea Public Tissue Bank, Seongnam-si, South Korea.
⁴ Department of Rheumatology, Hanyang University Hospital for Rheumatic Diseases, Seoul, South Korea.
⁵ Department of Orthopaedic Surgery, Hanyang University Hospital, Seoul, South Korea.

PMID: 36342052
PMCID: PMC9680201
DOI: 10.1302/2046-3758.1111.BJR-2021-0548.R2

Abstract

Aims: To test the hypothesis that reseeded anterior cruciate ligament (ACL)-derived cells have a better ability to survive and integrate into tendon extracellular matrix (ECM) and accelerate the ligamentization process, compared to adipose-derived mesenchymal stem cells (ADMSCs).

Methods: Acellularized tibialis allograft tendons were used. Tendons were randomly reseeded with ACL-derived cells or ADMSCs. ACL-derived cells were harvested and isolated from remnants of ruptured ACLs during reconstruction surgery and cultured at passage three. Cell suspensions (200 µl) containing 2 × 10⁶ ACL-derived cells or ADMSCs were prepared for the purpose of reseeding. At days 1, 3, and 7 post-reseeding, graft composites were assessed for repopulation with histological and immunohistochemical analysis. Matrix protein contents and gene expression levels were analyzed.

Results: In the graft reseeded with ACL-derived cells, a large number of elongated cells that integrated into the matrix were evident at day 3 and day 7. However, in the graft reseeded with ADMSCs, only a small number of elongated cells were found integrated into the matrix. Immunofluorescence for Ki-67 and type I collagen confirmed the pronounced production of type I collagen by Ki-67-positive ACL-derived cells integrated into the ECM. A messenger RNA (mRNA) expression assay demonstrated significantly higher gene expression levels of types I (p = 0.013) and III (p = 0.050) collagen in the composites reseeded with ACL-derived cells than ADMSCs.

Conclusion: ACL-derived cells, when reseeded to acellularized tendon graft, demonstrated earlier better survival and integration in the tendon ECM and resulted in higher gene expression levels of collagen, which may be essential to the normal ligamentization process compared to ADMSCs.Cite this article: Bone Joint Res 2022;11(11):777-786.

Keywords: ACL-derived cells; ADMSC; Acellularized tendon allograft; Ligamentization; RNA; Recellularization; adipose-derived mesenchymal stem cells; anterior cruciate ligament (ACL); collagen; gene expression; mRNA; tendon allografts; tendon grafts; tendons; type I collagen.