Skip to main content


Log in

Simvastatin promotes rat Achilles tendon-bone interface healing by promoting osteogenesis and chondrogenic differentiation of stem cells

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript


To investigate the effect and mechanism of simvastatin on cell components of tendon-bone healing interface. The tendon-bone healing model was established by inserting the end of the Achilles tendon into the tibial tunnel on 24 rats, and simvastatin was used locally at the tendon-bone interface. Healing was evaluated at 8 weeks by mechanical testing, micro-CT, and qualitative histology including H&E, Toluidine blue, and immunohistochemical staining. In vitro, bone marrow stromal cells (BMSCs) and tendon-derived mesenchymal stem cells (TDSCs) underwent osteogenic and chondrogenic differentiation respectively by plate co-culture. An analysis was performed on days 7 and 14 of cell differentiation. Biomechanical testing demonstrated a significant increase in maximum stiffness in the simvastatin-treated group. Micro-CT analysis showed that the bone tunnels in the simvastatin group were smaller in diameter and had higher bone density. H&E and Toluidine blue staining demonstrated that tendon-bone healing was significantly greater with better tissue arrangement and more extracellular matrix in the simvastatin-treated group than that in the control group, and immunohistochemical staining showed the expression of VEGF in simvastatin group was significantly higher. Histological staining and RT-PCR confirmed that simvastatin could promote the differentiation of co-cultured BMSCs and TDSCs into osteoblasts and chondroblasts, respectively. The effect of promoting osteogenic differentiation was more tremendous at 14 days, while its effect on promoting chondroblast differentiation was more evident on the 7th day of differentiation. In conclusion, local administration of simvastatin can promote the tendon-bone healing by enhancing neovascularization, chondrogenesis, and osteogenesis in different stages of the tendon-bone healing process.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others


  • Asai S, Otsuru S, Candela ME, Cantley L, Uchibe K, Hofmann TJ, Zhang K, Wapner KL, Soslowsky LJ, Horwitz EMJSC (2014) Tendon progenitor cells in injured tendons have strong chondrogenic potential: the CD105‐negative subpopulation induces chondrogenic degeneration. 32

  • Ball SG, Shuttleworth AC, Kielty CM (2004) Direct cell contact influences bone marrow mesenchymal stem cell fate. Int J Biochem Cell Biol 36:714–727

    Article  CAS  Google Scholar 

  • Bamba Y, Shofuda T, Kanematsu D, Nonaka M, Yamasaki M, Okano H, Kanemura Y (2014) Differentiation, polarization, and migration of human induced pluripotent stem cell-derived neural progenitor cells co-cultured with a human glial cell line with radial glial-like characteristics. Biochem Biophys Res Commun 447:683–688

    Article  CAS  Google Scholar 

  • Beri A, Dwamena FC, Dwamena BA (2009) Association between statin therapy and tendon rupture: a case-control study. J Cardiovasc Pharmacol 53:401–404

    Article  CAS  Google Scholar 

  • Bitto A, Minutoli L, Altavilla D, Polito F, Fiumara T, Marini H, Galeano M, Calo M, Lo Cascio P, Bonaiuto M, Migliorato A, Caputi AP, Squadrito F (2008) Simvastatin enhances VEGF production and ameliorates impaired wound healing in experimental diabetes. Pharmacol Res 57:159–169

    Article  CAS  Google Scholar 

  • Cai J, Wang J, Ye K, Li D, Ai C, Sheng D, Jin W, Liu X, Zhi Y, Jiang J, Chen J, Mo X, Chen S (2018) Dual-layer aligned-random nanofibrous scaffolds for improving gradient microstructure of tendon-to-bone healing in a rabbit extra-articular model. Int J Nanomedicine 13:3481–3492

    Article  CAS  Google Scholar 

  • Canseco JA, Kojima K, Penvose AR, Ross JD, Obokata H, Gomoll AH, Vacanti CA (2012) Effect on ligament marker expression by direct-contact co-culture of mesenchymal stem cells and anterior cruciate ligament cells. Tissue Eng Part A 18:2549–2558

    Article  CAS  Google Scholar 

  • Contractor T, Beri A, Gardiner JC, Tang X, Dwamena FC (2015) Is statin use associated with tendon rupture? A population-based retrospective cohort analysis. Am J Ther 22:377–381

    Article  Google Scholar 

  • Dimmen S, Nordsletten L, Engebretsen L, Steen H, Madsen JE (2009) The effect of parecoxib and indometacin on tendon-to-bone healing in a bone tunnel: an experimental study in rats. J Bone Joint Surg Br 91:259–263

    Article  CAS  Google Scholar 

  • Eliasson P, Dietrich-Zagonel F, Lundin AC, Aspenberg P, Wolk A, Michaelsson K (2019) Statin treatment increases the clinical risk of tendinopathy through matrix metalloproteinase release - a cohort study design combined with an experimental study. Sci Rep 9:17958

    Article  Google Scholar 

  • Eliasson P, Svensson RB, Giannopoulos A, Eismark C, Kjaer M, Schjerling P, Heinemeier KM (2017) Simvastatin and atorvastatin reduce the mechanical properties of tendon constructs in vitro and introduce catabolic changes in the gene expression pattern. PLoS ONE 12:e0172797

    Article  Google Scholar 

  • Font Tellado S, Balmayor ER, Van Griensven M (2015) Strategies to engineer tendon/ligament-to-bone interface: biomaterials, cells and growth factors. Adv Drug Deliv Rev 94:126–140

    Article  CAS  Google Scholar 

  • Hjorthaug GA, Soreide E, Nordsletten L, Madsen JE, Reinholt FP, Niratisairak S, Dimmen S (2018) Negative effect of zoledronic acid on tendon-to-bone healing. Acta Orthop 89:360–366

    Article  Google Scholar 

  • Ibrahim N, Mohamed N, Shuid AN (2013) Update on statins: hope for osteoporotic fracture healing treatment. Curr Drug Targets 14:1524–1532

    Article  CAS  Google Scholar 

  • Im GI, Kim TK (2020) Stem cells for the regeneration of tendon and ligament: a perspective. 13:335–341

  • Karaoglu S, Celik C, Korkusuz P (2009) The effects of bone marrow or periosteum on tendon-to-bone tunnel healing in a rabbit model. Knee Surg Sports Traumatol Arthrosc 17:170–178

    Article  Google Scholar 

  • Koellensperger E, Gramley F, Preisner F, Leimer U, Germann G, Dexheimer V (2014) Alterations of gene expression and protein synthesis in co-cultured adipose tissue-derived stem cells and squamous cell-carcinoma cells: consequences for clinical applications. Stem Cell Res Ther 5:65

    Article  Google Scholar 

  • Kuang GM, Yau WP, Lu WW, Chiu KY (2013) Use of a strontium-enriched calcium phosphate cement in accelerating the healing of soft-tissue tendon graft within the bone tunnel in a rabbit model of anterior cruciate ligament reconstruction. 95-B:923–928

  • Lee JI, Sato M, Kim HW, Mochida J (2011) Transplantatation of scaffold-free spheroids composed of synovium-derived cells and chondrocytes for the treatment of cartilage defects of the knee. Eur Cell Mater 22:275–290; discussion 290

  • Li H, Tang M, Liang H, Li Y, Wang J, Song Y, Zheng Y, Xi J, Zhang J, Hescheler J, Zhu M (2013) Coculture of embryonic ventricular myocytes and mouse embryonic stem cell enhance intercellular signaling by upregulation of connexin43. Cell Physiol Biochem 32:53–63

    Article  Google Scholar 

  • Lin TT, Lin CH, Chang CL, Chi CH, Chang ST, Sheu WH (2015) The effect of diabetes, hyperlipidemia, and statins on the development of rotator cuff disease: a nationwide, 11-year, longitudinal, population-based follow-up study. Am J Sports Med 43:2126–2132

    Article  Google Scholar 

  • Liu SH, Panossian V, Al-Shaikh R, Tomin E, Shepherd E, Finerman GA, Lane JM (1997) Morphology and matrix composition during early tendon to bone healing. Clin Orthop Relat Res 253–260

  • Lovati AB, Corradetti B, Cremonesi F, Bizzaro D, Consiglio AL (2012) Tenogenic differentiation of equine mesenchymal progenitor cells under indirect co-culture. Int J Artif Organs 35:996–1005

    Article  CAS  Google Scholar 

  • Lu J, Chamberlain CS, Ji ML, Saether EE, Leiferman EM, Li WJ, Vanderby R (2019) Tendon-to-bone healing in a rat extra-articular bone tunnel model: a comparison of fresh autologous bone marrow and bone marrow-derived mesenchymal stem cells. Am J Sports Med 47:2729–2736

    Article  Google Scholar 

  • Montero J, Manzano G, Albaladejo A (2014) The role of topical simvastatin on bone regeneration: a systematic review. J Clin Exp Dent 6:e286-290

    Article  Google Scholar 

  • Nagai H, Terada K, Watanabe G, Ueno Y, Aiba N, Shibuya T, Kawagoe M, Kameda T, Sato M, Senoo H, Sugiyama T (2002) Differentiation of liver epithelial (stem-like) cells into hepatocytes induced by coculture with hepatic stellate cells. Biochem Biophys Res Commun 293:1420–1425

    Article  CAS  Google Scholar 

  • Oka S, Matsumoto T, Kubo S, Matsushita T, Sasaki H, Nishizawa Y, Matsuzaki T, Saito T, Nishida K, Tabata Y, Kurosaka M, Kuroda R (2013) Local administration of low-dose simvastatin-conjugated gelatin hydrogel for tendon-bone healing in anterior cruciate ligament reconstruction. Tissue Eng Part A 19:1233–1243

    Article  CAS  Google Scholar 

  • Oryan A, Kamali A, Moshiri A (2015) Potential mechanisms and applications of statins on osteogenesis: current modalities, conflicts and future directions. J Control Release 215:12–24

    Article  CAS  Google Scholar 

  • Qin S, Wang W, Liu Z, Hua X, Fu SC, Dong F, Li A, Liu Z, Wang P, Dai L et al (2020) Fibrochondrogenic differentiation potential of tendon-derived stem/progenitor cells from human patellar tendon. 22

  • Reinoso RF, Sanchez Navarro A, Garcia MJ, Prous JR (2002) Preclinical pharmacokinetics of statins. Methods Find Exp Clin Pharmacol 24:593–613

    CAS  Google Scholar 

  • Rui YF, Lui PPY, Lee YW, Chan KM (2012) Higher BMP receptor expression and BMP-2-induced osteogenic differentiation in tendon-derived stem cells compared with bone-marrow-derived mesenchymal stem cells. 36:1099–1107

  • Schneider PR, Buhrmann C, Mobasheri A, Matis U, Shakibaei M (2011) Three-dimensional high-density co-culture with primary tenocytes induces tenogenic differentiation in mesenchymal stem cells. J Orthop Res 29:1351–1360

    Article  CAS  Google Scholar 

  • Schumm MA, Castellanos DA, Frydel BR, Sagen J (2003) Direct cell-cell contact required for neurotrophic effect of chromaffin cells on neural progenitor cells. Brain Res Dev Brain Res 146:1–13

    Article  CAS  Google Scholar 

  • Stein D, Lee Y, Schmid MJ, Killpack B, Genrich MA, Narayana N, Marx DB, Cullen DM, Reinhardt RA (2005) Local simvastatin effects on mandibular bone growth and inflammation. J Periodontol 76:1861–1870

    Article  CAS  Google Scholar 

  • Tai IC, Fu YC, Wang CK, Chang JK, Ho ML (2013) Local delivery of controlled-release simvastatin/PLGA/HAp microspheres enhances bone repair. Int J Nanomedicine 8:3895–3904

    Google Scholar 

  • Takahashi H, Tamaki H, Oyama M, Yamamoto N, Onishi H (2017) Time-dependent changes in the structure of calcified fibrocartilage in the rat Achilles tendon-bone interface with sciatic denervation. Anat Rec (hoboken) 300:2166–2174

    Article  CAS  Google Scholar 

  • Thylin MR, McConnell JC, Schmid MJ, Reckling RR, Ojha J, Bhattacharyya I, Marx DB, Reinhardt RA (2002) Effects of simvastatin gels on murine calvarial bone. J Periodontol 73:1141–1148

    Article  CAS  Google Scholar 

  • Wu T, Liu Y, Wang B, Li G (2014) The roles of mesenchymal stem cells in tissue repair and disease modification. Curr Stem Cell Res Ther 9:424–431

    Article  CAS  Google Scholar 

  • Wu T, Liu Y, Wang B, Sun Y, Xu J, Yuk-Wai LW, Xu L, Zhang J, Li G (2016) The use of cocultured mesenchymal stem cells with tendon-derived stem cells as a better cell source for tendon repair. Tissue Eng Part A 22:1229–1240

    Article  CAS  Google Scholar 

  • Xu K, Zhang Z, Chen M, Moqbel S, Wu LD (2020) Nesfatin-1 promotes the osteogenic differentiation of tendon-derived stem cells and the pathogenesis of heterotopic ossification in rat tendons via the mTOR pathway. 8:547342

  • Yeh WL, Lin SS, Yuan LJ, Lee KF, Lee MY, Ueng SW (2007) Effects of hyperbaric oxygen treatment on tendon graft and tendon-bone integration in bone tunnel: biochemical and histological analysis in rabbits. J Orthop Res 25:636–645

    Article  Google Scholar 

  • Yoshikawa T, Tohyama H, Katsura T, Kondo E, Kotani Y, Matsumoto H, Toyama Y, Yasuda K (2006) Effects of local administration of vascular endothelial growth factor on mechanical characteristics of the semitendinosus tendon graft after anterior cruciate ligament reconstruction in sheep. Am J Sports Med 34:1918–1925

    Article  Google Scholar 

  • Zhang L, Tran N, Chen HQ, Kahn CJ, Marchal S, Groubatch F, Wang X (2008) Time-related changes in expression of collagen types I and III and of tenascin-C in rat bone mesenchymal stem cells under co-culture with ligament fibroblasts or uniaxial stretching. Cell Tissue Res 332:101–109

    Article  CAS  Google Scholar 

  • Zhang P, Han F, Li Y, Chen J, Chen T, Zhi Y, Jiang J, Lin C, Chen S, Zhao P (2016) Local delivery of controlled-release simvastatin to improve the biocompatibility of polyethylene terephthalate artificial ligaments for reconstruction of the anterior cruciate ligament. Int J Nanomedicine 11:465–478

    Article  CAS  Google Scholar 

  • Zhang Y, Yu J, Zhang J, Hua Y (2019) Simvastatin with PRP promotes chondrogenesis of bone marrow stem cells in vitro and wounded rat Achilles tendon-bone interface healing in vivo. Am J Sports Med 47:729–739

    Article  Google Scholar 

  • Zhernasechanka HA, Isaikina YI, Filipovich TV, Liakh EG (2021) Osteogenic and chondrogenic differentiation potential of mesenchymal stem cells obtained from the bone marrow and placenta. 18:36–45

  • Zhou Y, Zhang J, Yang J, Narava M, Zhao G, Yuan T, Wu H, Zheng N, Hogan MV, Wang JH (2017) Kartogenin with PRP promotes the formation of fibrocartilage zone in the tendon-bone interface. J Tissue Eng Regen Med 11:3445–3456

    Article  CAS  Google Scholar 

Download references


This work was supported by grants from the National Natural Science Foundation of China (No. 81803275), the Fundamental Research Funds for the Central Universities (No. 2042017kf0086), and the Zhongnan Hospital of Wuhan University Science, Technology and Innovation Seed Fund (No. ZNPY2016017).

Author information

Authors and Affiliations



Qubo Ni, Liaobin Chen, and Jiayong Zhu designed this research; Qubo Ni, Jiayong Zhu, and Zhenyu Li performed the research; Qubo Ni, Jiayong Zhu, Bin Li, and Hui Wang analyzed the data; Jiayong Zhu, Liaobin Chen, and Qubo Ni wrote and revised the paper; and all authors approved the final manuscript.

Corresponding author

Correspondence to Liaobin Chen.

Ethics declarations

Ethics approval

All cell isolation and surgery procedures on experimental animals were carried out according to the Care and Use of Laboratory Animals guide and approved by the Institutional Animal Care and Use Committee of Wuhan University (IACUC Approval: 14016).

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ni, Q., Zhu, J., Li, Z. et al. Simvastatin promotes rat Achilles tendon-bone interface healing by promoting osteogenesis and chondrogenic differentiation of stem cells. Cell Tissue Res 391, 339–355 (2023).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: