Viable Tendon Neotissue from Adult Adipose-derived Multipotent Stromal Cells

Overview

Journal Front Bioeng Biotechnol

Date 2024 Jan 23

PMID 38260742

Authors

Takashi Taguchi

Mandi Lopez

Catherine Takawira

Affiliations

Soon will be listed here.

Abstract

Tendon healing is frequently prolonged, unpredictable, and results in poor tissue quality. Neotissue formed by adult multipotent stromal cells has the potential to guide healthy tendon tissue formation. The objective of this study was to characterize tendon neotissue generated by equine adult adipose-derived multipotent stromal cells (ASCs) on collagen type I (COLI) templates under 10% strain in a novel bioreactor. The tested hypothesis was that ASCs assume a tendon progenitor cell-like morphology, express tendon-related genes, and produce more organized extracellular matrix (ECM) in tenogenic versus stromal medium with perfusion and centrifugal fluid motion. Equine ASCs on COLI sponge cylinders were cultured in stromal or tenogenic medium within bioreactors during combined perfusion and centrifugal fluid motion for 7, 14, or 21 days under 10% strain. Viable cell distribution and number, tendon-related gene expression, and micro- and ultra-structure were evaluated with calcein-AM/EthD-1 staining, resazurin reduction, RT-PCR, and light, transmission, and scanning electron microscopy. Fibromodulin was localized with immunohistochemistry. Cell number and gene expression were compared between culture media and among culture periods ( < 0.05). Viable cells were distributed throughout constructs for up to 21 days of culture, and cell numbers were higher in tenogenic medium. Individual cells had a round or rhomboid shape with scant ECM in stromal medium in contrast to clusters of parallel, elongated cells surrounded by highly organized ECM in tenogenic medium after 21 days of culture. Transcription factor, extracellular matrix, and mature tendon gene expression profiles confirmed ASC differentiation to a tendon progenitor-like cell in tenogenic medium. Construct micro- and ultra-structure were consistent with tendon neotissue and fibromodulin was present in the ECM after culture in tenogenic medium. Long-term culture in custom bioreactors with combined perfusion and centrifugal tenogenic medium circulation supports differentiation of equine adult ASCs into tendon progenitor-like cells capable of neotissue formation.

References

Kim T, Sridharan I, Zhu B, Orgel J, Wang R . Effect of CNT on collagen fiber structure, stiffness assembly kinetics and stem cell differentiation. Mater Sci Eng C Mater Biol Appl. 2015; 49:281-289. PMC: 7225775. DOI: 10.1016/j.msec.2015.01.014. View

Ning L, Zhang Y, Zhang Y, Zhu M, Ding W, Jiang Y . Enhancement of Migration and Tenogenic Differentiation of Macaca Mulatta Tendon-Derived Stem Cells by Decellularized Tendon Hydrogel. Front Cell Dev Biol. 2021; 9:651583. PMC: 8111289. DOI: 10.3389/fcell.2021.651583. View

Snedeker J, Foolen J . Tendon injury and repair - A perspective on the basic mechanisms of tendon disease and future clinical therapy. Acta Biomater. 2017; 63:18-36. DOI: 10.1016/j.actbio.2017.08.032. View

Geburek F, Roggel F, van Schie H, Beineke A, Estrada R, Weber K . Effect of single intralesional treatment of surgically induced equine superficial digital flexor tendon core lesions with adipose-derived mesenchymal stromal cells: a controlled experimental trial. Stem Cell Res Ther. 2017; 8(1):129. PMC: 5460527. DOI: 10.1186/s13287-017-0564-8. View

Zhao Y, Zhang S, Zhou J, Wang J, Zhen M, Liu Y . The development of a tissue-engineered artery using decellularized scaffold and autologous ovine mesenchymal stem cells. Biomaterials. 2009; 31(2):296-307. DOI: 10.1016/j.biomaterials.2009.09.049. View

Rossdale P, Hopes R, Digby N, Offord K . Epidemiological study of wastage among racehorses 1982 and 1983. Vet Rec. 1985; 116(3):66-9. DOI: 10.1136/vr.116.3.66. View

Korcari A, Muscat S, McGinn E, Buckley M, Loiselle A . Depletion of Scleraxis-lineage cells during tendon healing transiently impairs multi-scale restoration of tendon structure during early healing. PLoS One. 2022; 17(10):e0274227. PMC: 9565440. DOI: 10.1371/journal.pone.0274227. View

Kannus P, Jozsa L . Histopathological changes preceding spontaneous rupture of a tendon. A controlled study of 891 patients. J Bone Joint Surg Am. 1991; 73(10):1507-25. View

Taye N, Karoulias S, Hubmacher D . The "other" 15-40%: The Role of Non-Collagenous Extracellular Matrix Proteins and Minor Collagens in Tendon. J Orthop Res. 2019; 38(1):23-35. PMC: 6917864. DOI: 10.1002/jor.24440. View

10.

Engebretson B, Mussett Z, Sikavitsas V . Tenocytic extract and mechanical stimulation in a tissue-engineered tendon construct increases cellular proliferation and ECM deposition. Biotechnol J. 2016; 12(3). DOI: 10.1002/biot.201600595. View

11.

Duan W, Haque M, Kearney M, Lopez M . Collagen and Hydroxyapatite Scaffolds Activate Distinct Osteogenesis Signaling Pathways in Adult Adipose-Derived Multipotent Stromal Cells. Tissue Eng Part C Methods. 2017; 23(10):592-603. PMC: 5653142. DOI: 10.1089/ten.TEC.2017.0078. View

12.

Schneider P, Buhrmann C, Mobasheri A, Matis U, Shakibaei M . Three-dimensional high-density co-culture with primary tenocytes induces tenogenic differentiation in mesenchymal stem cells. J Orthop Res. 2011; 29(9):1351-60. DOI: 10.1002/jor.21400. View

13.

Yang F, Zhang A, Richardson D . Regulation of the tenogenic gene expression in equine tenocyte-derived induced pluripotent stem cells by mechanical loading and Mohawk. Stem Cell Res. 2019; 39:101489. PMC: 7082636. DOI: 10.1016/j.scr.2019.101489. View

14.

Dyson S . Medical management of superficial digital flexor tendonitis: a comparative study in 219 horses (1992-2000). Equine Vet J. 2004; 36(5):415-9. DOI: 10.2746/0425164044868422. View

15.

Williams R, Harkins L, Hammond C, Wood J . Racehorse injuries, clinical problems and fatalities recorded on British racecourses from flat racing and National Hunt racing during 1996, 1997 and 1998. Equine Vet J. 2001; 33(5):478-86. DOI: 10.2746/042516401776254808. View

16.

Zhang B, Luo Q, Deng B, Morita Y, Ju Y, Song G . Construction of tendon replacement tissue based on collagen sponge and mesenchymal stem cells by coupled mechano-chemical induction and evaluation of its tendon repair abilities. Acta Biomater. 2018; 74:247-259. DOI: 10.1016/j.actbio.2018.04.047. View

17.

Kuo C, Tuan R . Mechanoactive tenogenic differentiation of human mesenchymal stem cells. Tissue Eng Part A. 2008; 14(10):1615-27. DOI: 10.1089/ten.tea.2006.0415. View

18.

Cardwell R, Dahlgren L, Goldstein A . Electrospun fibre diameter, not alignment, affects mesenchymal stem cell differentiation into the tendon/ligament lineage. J Tissue Eng Regen Med. 2012; 8(12):937-45. DOI: 10.1002/term.1589. View

19.

Grier W, Moy A, Harley B . Cyclic tensile strain enhances human mesenchymal stem cell Smad 2/3 activation and tenogenic differentiation in anisotropic collagen-glycosaminoglycan scaffolds. Eur Cell Mater. 2017; 33:227-239. PMC: 5453510. DOI: 10.22203/eCM.v033a14. View

20.

Kaji D, Howell K, Balic Z, Hubmacher D, Huang A . Tgfβ signaling is required for tenocyte recruitment and functional neonatal tendon regeneration. Elife. 2020; 9. PMC: 7324157. DOI: 10.7554/eLife.51779. View