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. 2022 May 13;17(1):266.
doi: 10.1186/s13018-022-03152-y.

Upregulation of Runt related transcription factor 1 (RUNX1) contributes to tendon-bone healing after anterior cruciate ligament reconstruction using bone mesenchymal stem cells

Kai Kang  1 , Qian Geng  1 , Lukuan Cui  1 , Lijie Wu  1 , Lei Zhang  1 , Tong Li  1 , Qian Zhang  1 , Shijun Gao  2
Affiliations

Upregulation of Runt related transcription factor 1 (RUNX1) contributes to tendon-bone healing after anterior cruciate ligament reconstruction using bone mesenchymal stem cells

Kai Kang et al. J Orthop Surg Res. .

Abstract

Background: Anterior cruciate ligament (ACL) injury could lead to functional impairment along with disabilities. ACL reconstruction often fails owing to the regeneration failure of tendon-bone interface. Herein, we aimed to investigate the effects of Runt related transcription factor 1 (RUNX1) on tendon-bone healing after ACL reconstruction using bone mesenchymal stem cells (BMSCs).

Methods: BMSCs were isolated from the marrow cavity of rat femur, followed by the modification of RUNX1 with lentiviral system. Then, an ACL reconstruction model of rats was established with autografts.

Results: Results of flow cytometry exhibited positive-antigen CD44 and CD90, as well as negative-antigen CD34 and CD45 of the BMSCs. Then, we found that RUNX1-upregulated BMSCs elevated the decreased biomechanical strength of the tendon grafts after ACL reconstruction. Moreover, based on the histological observation, upregulation of RUNX1 was linked with better recovery around the bone tunnel, a tighter tendon-bone interface, and more collagen fibers compared to the group of BMSCs infected with LV-NC. Next, RUNX1-upregulated BMSCs promoted osteogenesis after ACL reconstruction, as evidenced by the mitigation of severe loss and erosion of the cartilage and bone in the tibial and femur area, as well as the increased number of osteoblasts identified by the upregulation of alkaline phosphatase, osteocalcin, and osteopontin in the tendon-bone interface.

Conclusion: Elevated expression of RUNX1 contributed to tendon-bone healing after ACL reconstruction using BMSCs.

Keywords: Anterior cruciate ligament reconstruction; Bone mesenchymal stem cells; Runt related transcription factor 1; Tendon–bone healing.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Phenotype identification of BMSCs. A Primary BMSCs exhibited a long spindle shape (100× magnification). B BMSCs surface antigen identification by flow cytometry. C BMSCs infected with LV-NC or LV-RUNX1 by fluorescence microscopy (100× magnification). D The mRNA and protein expression of RUNX1 in BMSCs. ##p < 0.01
Fig. 2
Fig. 2
RUNX1-upregulated BMSCs elevates biomechanical strength after ACL reconstruction. A–F Procedures for applying the BMSCs infected with LV-NC or LV-RUNX1 sheet to the bone tunnel during ACL reconstruction. A, B Harvest the native tendon as the tendon graft. C, D Create the femoral and tibial bone tunnels. E Insert the tendon graft into both bone tunnels. F Fix the tendon graft with suture tied over the neighboring periosteum. G Ultimate load of the tendon graft. H Stiffness of the tendon graft. #p < 0.05, ##p < 0.01; NS not significant
Fig. 3
Fig. 3
RUNX1-upregulated BMSCs accelerates the healing of tendon–bone interface after ACL reconstruction. A Fluorescence imaging of tendon–bone interface (400× magnification). B H&E staining of the interface (400× magnification). C Masson’s trichrome staining of the interface (400× magnification). B bone, IF interface, T tendon
Fig. 4
Fig. 4
RUNX1-upregulated BMSCs promotes osteogenesis after ACL reconstruction. A Micro-CT images of the knee joints. B BMD and BV/TV. C ALP-stained cells in tendon to bone interface (400× magnification). Black arrows indicated the ALP-positive cells. D, E The expression of OCN and OPN was assessed by immunohistochemistry (800× magnification). Black arrows indicated the OCN and OPN-positive cells in tendon to bone interface. #p < 0.05, ##p < 0.01; NS not significant
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References

    1. Rothrauff BB, Tuan RS. Cellular therapy in bone-tendon interface regeneration. Organogenesis. 2014;10(1):13–28. doi: 10.4161/org.27404. - DOI - PMC - PubMed
    1. Killian ML. Growth and mechanobiology of the tendon–bone enthesis. Semin Cell Dev Biol. 2021;123:64–73. doi: 10.1016/j.semcdb.2021.07.015. - DOI - PMC - PubMed
    1. Cottrell JA, Turner JC, Arinzeh TL, O'Connor JP. The biology of bone and ligament healing. Foot Ankle Clin. 2016;21(4):739–761. doi: 10.1016/j.fcl.2016.07.017. - DOI - PubMed
    1. Hjorthaug GA, Søreide E, Nordsletten L, Madsen JE, Reinholt FP, Niratisairak S, et al. Negative effect of zoledronic acid on tendon-to-bone healing. Acta Orthop. 2018;89(3):360–366. doi: 10.1080/17453674.2018.1440189. - DOI - PMC - PubMed
    1. Ryu K, Saito M, Kurosaka D, Kitasato S, Omori T, Hayashi H, et al. Enhancement of tendon–bone interface healing and graft maturation with cylindrical titanium-web (TW) in a miniature swine anterior cruciate ligament reconstruction model: histological and collagen-based analysis. BMC Musculoskelet Disord. 2020;21(1):198. doi: 10.1186/s12891-020-03199-0. - DOI - PMC - PubMed

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