Open Access

The role of antimiR-26a-5p/biphasic calcium phosphate in repairing rat femoral defects

  • Authors:
    • Xiaoyan Yuan
    • Lu Han
    • Hai Lin
    • Zeyou Guo
    • Yanling Huang
    • Shasha Li
    • Ting Long
    • Wei Tang
    • Weidong Tian
    • Jie Long
  • View Affiliations

  • Published online on: June 20, 2019     https://doi.org/10.3892/ijmm.2019.4249
  • Pages: 857-870
  • Copyright: © Yuan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Although miRNAs have been implicated in the osteogenic differentiation of stem cells, their role in bone repair and reconstruction in tissue‑engineered bone grafts remains unclear. We previously reported that microRNA (miR)‑26a‑5p inhibited the osteogenic differentiation of adipose‑derived mesenchymal stem cells (ADSCs), and that antimiR‑26a‑5p exerted the opposite effect. In the present study, the role of miR‑26a‑5p‑ and antimiR‑26a‑5p‑modified ADSCs combined with biphasic calcium phosphate (BCP) scaffolds was evaluated in a rat femur defect model. The aim of the present study was to improve the understanding of the role of miR‑26a‑5p in bone regeneration in vivo, as well as to provide a new method to optimize the osteogenic ability of BCPs. ADSCs were infected with Lv‑miR‑26a‑5p, Lv‑miR‑NC, Lv‑antimiR‑26a‑5p or Lv‑antimiR‑NC respectively, and then combined with BCP scaffolds to repair rat femoral defects. Using X‑rays, micro‑computed tomography and histology at 2, 4, and 8 weeks postoperatively, the quantity and rate of bone regeneration were analyzed, revealing that they were the highest in animals treated with antimiR‑26a‑5p and the lowest in the miR‑26a‑5p treatment group. The expression levels of osteocalcin, collagen I, Runt‑related transcription factor 2, Wnt family member 5A and calmodulin‑dependent protein kinase II proteins were positively correlated with the bone formation rate. Taken together, the present results demonstrated that miR‑26a‑5p inhibited bone formation while antimiR‑26a‑5p accelerated bone formation via the Wnt/Ca2+ signaling pathway. Therefore, antimiR‑26a‑5p‑modified ADSCs combined with BCP scaffolds may be used to construct an effective tissue‑engineering bone graft for bone repair and reconstruction.

References

1 

Lee JE, Kim MB, Han DH, Pyo SH and Lee YH: One-barrel microsurgical fibula flap for reconstruction of large defects of the femur. Ann Plast Surg. 80:373–378. 2018. View Article : Google Scholar : PubMed/NCBI

2 

Mohseni M, Jahandideh A, Abedi G, Akbarzadeh A and Hesaraki S: Assessment of tricalcium phosphate/collagen (TCP/collagene) nanocomposite scaffold compared with hydroxyapatite (HA) on healing of segmental femur bone defect in rabbits. Artif Cells Nanomed Biotechnol. 46:242–249. 2018. View Article : Google Scholar

3 

Sakkas A, Schramm A, Winter K and Wilde F: Risk factors for post-operative complications after procedures for autologous bone augmentation from different donor sites. J Craniomaxillofac Surg. 46:312–322. 2018. View Article : Google Scholar

4 

Burk T, Del Valle J, Finn RA and Phillips C: Maximum quantity of bone available for harvest from the anterior iliac crest, posterior iliac crest, and proximal tibia using a standardized surgical approach: A cadaveric study. J Oral Maxillofac Surg. 74:2532–2548. 2016. View Article : Google Scholar : PubMed/NCBI

5 

Aponte-Tinao LA, Albergo JI, Ayerza MA, Muscolo DL, Ing FM and Farfalli GL: What Are the complications of allograft reconstructions for sarcoma resection in children younger than 10 years at long-term followup? Clin Orthop Relat Res. 476:548–555. 2018. View Article : Google Scholar : PubMed/NCBI

6 

Khodakaram-Tafti A, Mehrabani D, Shaterzadeh-Yazdi H, Zamiri B and Omidi M: Tissue engineering in maxillary bone defects. World J Plast Surg. 7:3–11. 2018.PubMed/NCBI

7 

Diomede F, Gugliandolo A, Cardelli P, Merciaro I, Ettorre V, Traini T, Bedini R, Scionti D, Bramanti A, Nanci A, et al: Three-dimensional printed PLA scaffold and human gingival stem cell-derived extracellular vesicles: A new tool for bone defect repair. Stem Cell Res Ther. 9:1042018. View Article : Google Scholar : PubMed/NCBI

8 

Reichert JC, Saifzadeh S, Wullschleger ME, Epari DR, Schütz MA, Duda GN, Schell H, van Griensven M, Redl H and Hutmacher DW: The challenge of establishing preclinical models for segmental bone defect research. Biomaterials. 30:2149–2163. 2009. View Article : Google Scholar : PubMed/NCBI

9 

Le BQ, Nurcombe V, Cool SM, van Blitterswijk CA, de Boer J and LaPointe VLS: The components of bone and what they can teach us about regeneration. Materials (Basel). 11. pp. E142017, View Article : Google Scholar

10 

Horch RE, Beier JP, Kneser U and Arkudas A: Successful human long-term application of in situ bone tissue engineering. J Cell Mol Med. 18:1478–1485. 2014. View Article : Google Scholar : PubMed/NCBI

11 

Pennesi G, Scaglione S, Giannoni P and Quarto R: Regulatory influence of scaffolds on cell behavior: How cells decode bioma-terials. Curr Pharm Biotechnol. 12:151–159. 2011. View Article : Google Scholar

12 

Uzeda MJ, de Brito Resende RF, Sartoretto SC, Alves ATNN, Granjeiro JM and Calasans-Maia MD: Randomized clinical trial for the biological evaluation of two nanostructured biphasic calcium phosphate biomaterials as a bone substitute. Clin Implant Dent Relat Res. 19:802–811. 2017. View Article : Google Scholar : PubMed/NCBI

13 

Yun PY, Kim YK, Jeong KI, Park JC and Choi YJ: Influence of bone morphogenetic protein and proportion of hydroxyapatite on new bone formation in biphasic calcium phosphate graft: Two pilot studies in animal bony defect model. J Craniomaxillofac Surg. 42:1909–1917. 2014. View Article : Google Scholar : PubMed/NCBI

14 

Wang S, Zhang Z, Zhao J, Zhang X, Sun X, Xia L, Chang Q, Ye D and Jiang X: Vertical alveolar ridge augmentation with beta-tricalcium phosphate and autologous osteoblasts in canine mandible. Biomaterials. 30:2489–2498. 2009. View Article : Google Scholar : PubMed/NCBI

15 

Shuang Y, Yizhen L, Zhang Y, Fujioka-Kobayashi M, Sculean A and Miron RJ: In vitro characterization of an osteoinductive biphasic calcium phosphate in combination with recombinant BMP2. BMC Oral Health. 17:352016. View Article : Google Scholar : PubMed/NCBI

16 

Dragonas P, Palin C, Khan S, Gajendrareddy PK and Weiner WD: Complications associated with the use of recombinant human bone morphogenic protein-2 in ridge augmentation: A case report. J Oral Implantol. 43:351–359. 2017. View Article : Google Scholar : PubMed/NCBI

17 

Uludag H, D'Augusta D, Palmer R, Timony G and Wozney J: Characterization ofrhBMP-2 pharmacokinetics implanted with biomaterial carriers in the rat ectopic model. J Biomed Mater Res. 46:193–202. 1999. View Article : Google Scholar : PubMed/NCBI

18 

Arinzeh TL, Tran T, Mcalary J and Daculsi G: A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation. Biomaterials. 26:3631–3638. 2005. View Article : Google Scholar

19 

van Esterik FA, Zandieh-Doulabi B, Kleverlaan CJ and Klein-Nulend J: Enhanced Osteogenic and Vasculogenic differentiation potential of human adipose stem cells on biphasic calcium phosphate scaffolds in fibrin gels. Stem Cells Int. 2016:19342702016. View Article : Google Scholar : PubMed/NCBI

20 

Lian JB, Stein GS, van Wijnen AJ, Stein JL, Hassan MQ, Gaur T and Zhang Y: MicroRNA control of bone formation and homeostasis. Nat Rev Endocrinol. 8:212–227. 2012. View Article : Google Scholar : PubMed/NCBI

21 

Zuo B, Zhu J, Li J, Wang C, Zhao X, Cai G, Li Z, Peng J, Wang P, Shen C, et al: MicroRNA-103a functions as a mechanosensitive microRNA to inhibit bone formation through targeting Runx2. J Bone Miner Res. 30:330–345. 2015. View Article : Google Scholar

22 

Hupkes M, Sotoca AM, Hendriks JM, van Zoelen EJ and Dechering KJ: Micro-RNA miR-378 promotes BMP2-induced osteogenic differentiation of mesenchymal progenitor cells. BMC Mol Biol. 15:12014. View Article : Google Scholar

23 

Wang Q, Cai J, Cai XH and Chen L: MiR-346 regulates osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by targeting the Wnt/β-catenin pathway. PLoS One. 8:pp. e722662013, View Article : Google Scholar

24 

Bartel DP: MicroRNAs: Target recognition and regulatory functions. Cell. 136:215–233. 2009. View Article : Google Scholar : PubMed/NCBI

25 

Li JW, Hu C, Han L, Liu L, Jing W, Tang W, Tian WD and Long J: MiR-154-5p regulates osteogenic differentiation of adipose-derived mesenchymal stem cells under tensile stress through the Wnt/PCP pathway by targeting Wnt11. Bone. 78:130–141. 2015. View Article : Google Scholar : PubMed/NCBI

26 

Wu T, Zhou H, Hong Y, Li J, Jiang X and Huang H: MiR-30 family members negatively regulate osteoblast differentiation. J Biol Chem. 287:7503–7511. 2012. View Article : Google Scholar : PubMed/NCBI

27 

Li Z, Hassan MQ, Volinia S, van Wijnen AJ, Stein JL, Croce CM, Lian JB and Stein GS: A microRNA signature for a BMP2-induced osteoblast lineage commitment program. Proc Natl Acad Sci USA. 105:13906–13911. 2008. View Article : Google Scholar : PubMed/NCBI

28 

Suh JS, Lee JY, Choi YS, Chong PC and Park YJ: Peptide-mediated intracellular delivery of miRNA-29b for osteogenic stem cell differentiation. Biomaterials. 34:4347–4359. 2013. View Article : Google Scholar : PubMed/NCBI

29 

Li S, Hu C, Li J, Liu L, Jing W, Tang W, Tian W and Long J: Effect of miR-26a-5p on the Wnt/Ca(2+) Pathway and Osteogenic differentiation of mouse Adipose-Derived mesenchymal stem cells. Calcif Tissue Int. 99:174–186. 2016. View Article : Google Scholar : PubMed/NCBI

30 

Choudhery MS, Badowski M, Muise A and Harris DT: Comparison of human mesenchymal stem cells derived from adipose and cord tissue. Cytotherapy. 15:330–343. 2013. View Article : Google Scholar : PubMed/NCBI

31 

Ansari S, Diniz IM, Chen C, Sarrion P, Tamayol A, Wu BM and Moshaverinia A: Human periodontal ligament- and gingiva-derived mesenchymal stem cells promote nerve regeneration when encapsulated in alginate/hyaluronic acid 3D scaffold. Adv Healthc Mater. Oct 27–2017, Epub ahead of print. PubMed/NCBI

32 

Chen Y, Wang J, Zhu XD, Tang ZR, Yang X, Tan YF, Fan YJ and Zhang XD: Enhanced effect of β-tricalcium phosphate phase on neovascularizationof porous calcium phosphate ceramics: In vitro and in vivo evidence. Acta Biomater. 11:435–448. 2015. View Article : Google Scholar

33 

Huang L, Zhou B, Wu H, Zheng L and Zhao JM: Effect of apatite formation of Biphasic calcium phosphate (BCP) on the osteoblastogenesis using simulated body fluid with or without bovine serum albumin. Mater Sci Eng C Mater Biol Appl. 70:955–961. 2017. View Article : Google Scholar

34 

Meng YB, Li X, Li ZY, Zhao J, Yuan XB, Ren Y, Cui ZD, Liu YD and Yang XJ: MicroRNA-21 promotes osteogenic differentiation of mesenchymal stem cells by the PI3K/β-catenin pathway. J Orthop Res. 33:957–964. 2015. View Article : Google Scholar : PubMed/NCBI

35 

Liao YH, Chang YH, Sung LY, Li KC, Yeh CL, Yen TC, Hwang SM, Lin KJ and Hu YC: Osteogenic differentiation of adipose-derived stem cells and calvarial defect repair using baculovirus-mediated co-expression of BMP-2 and miR-148b. Biomaterials. 35:4901–4910. 2014. View Article : Google Scholar : PubMed/NCBI

36 

Huang S, Wang S, Bian C, Yang Z, Zhou H, Zeng Y, Li H, Han Q and Zhao RC: Upregulation of miR-22 promotes osteogenic differentiation and inhibits adipogenic differentiation of human adipose tissue-derived mesenchymal stem cells by repressing HDAC6 protein expression. Stem Cells Dev. 21:2531–2540. 2012. View Article : Google Scholar : PubMed/NCBI

37 

Wang Y, Li YP, Paulson C, Shao JZ, Zhang X, Wu M and Chen W: Wnt and the Wnt signaling pathway in bone development and disease. Front Biosci (Landmark Ed). 19:379–407. 2014. View Article : Google Scholar

38 

Martineau X, Abed É, Martel-Pelletier J, Pelletier JP and Lajeunesse D: Alteration of Wnt5a expression and of the non-canonical Wnt/PCP and Wnt/PKC-Ca2+ pathways in human osteoarthritis osteoblasts. PLoS One. 12:pp. e01807112017, View Article : Google Scholar

39 

Lacroix D, Chateau A, Ginebra MP and Planell JA: Micro-finite element models of bone tissue-engineering scaffolds. Biomaterials. 27:5326–5334. 2006. View Article : Google Scholar : PubMed/NCBI

40 

Bouler JM, Pilet P, Gauthier O and Verron E: Biphasic calcium phosphate ceramics for bone reconstruction: A reviewof biological response. Acta Biomater. 53:1–12. 2017. View Article : Google Scholar : PubMed/NCBI

41 

Tang XH, Mao LX, Liu JQ, Yang Z, Zhang W, Shu MJ, Hu NT, Jiang LY and Fang B: Fabrication, characterization and cellular biocompatibility of porous biphasic calcium phosphate bioceramic scaffolds with different pore sizes. Ceram Int. 42:15311–15318. 2016. View Article : Google Scholar

42 

Wu Y, Xia L, Zhou Y, Ma W, Zhang N, Chang J, Lin KL, Xu YJ and Jiang XQ: Evaluation of osteogenesis and angiogenesis of icariin loaded on micro/nanohybrid structured hydroxyapatite granules as a local drug delivery system forfemoral defect repair. J Mater Chem B. 3:4871–4883. 2015. View Article : Google Scholar

43 

Ebrahimi M, Botelho MG and Dorozhkin SV: Biphasic calcium phosphates bioceramics (HA/TCP): Concept, physicochemical properties and the impact of standardization of study protocols in biomaterials research. Mater Sci Eng C Mater Biol Appl. 71:1293–1312. 2017. View Article : Google Scholar

44 

Jensen SS, Bornstein MM, Dard M, Bosshardt DD and Buser D: Comparative study of biphasic calcium phosphates with different HA/TCP ratios in mandibular bone defects. Along-term histo-morphometric study in minipigs. J Biomed Mater Res B Appl Biomater. 90:171–181. 2009.

45 

Zhu Y, Zhang K, Zhao R, Ye X, Chen X, Xiao Z, Yang X, Zhu X, Zhang K, Fan Y and Zhang X: Bone regeneration with micro/nano hybrid-structured biphasic calcium phosphate bioceramics at segmental bone defect and the induced immunoregulation of MSCs. Biomaterials. 147:133–144. 2017. View Article : Google Scholar : PubMed/NCBI

46 

Ng AM, Tan KK, Phang MY, Aziyati O, Tan GH, Isa MR, Aminuddin BS, Naseem M, Fauziah O and Ruszymah BH: Differential osteogenic activity of osteoprogenitor cells on HA and TCP/HA scaffold of tissue engineered bone. J Biomed Mater Res A. 85:301–312. 2008. View Article : Google Scholar

47 

Ebrahimian-Hosseinabadi M, Etemadifar M and Ashrafizadeh F: Effects of nanobiphasic calcium phosphate composite on bioactivity and osteoblast cell behavior in tissue engineering applications. J Med Signals Sens. 6:237–242. 2016.PubMed/NCBI

48 

Huang J, Ten E, Liu G, Finzen M, Yu W, Lee JS, Saiz E and Tomsia AP: Biocomposites of pHEMA with HA/beta-TCP (60/40) for bone tissue engineering: Swelling, hydrolytic degradation, and in vitro behavior. Polymer (Guildf). 54:1197–1207. 2013. View Article : Google Scholar

49 

Daculsi G, Bouler JM and LeGeros RZ: Adaptive crystal formation in normal and pathological calcifications in synthetic calcium phosphate and related biomaterials. Int Rev Cytol. 172:129–191. 1997. View Article : Google Scholar : PubMed/NCBI

50 

Monchau F, Lefevre A, Descamps M, Belquinmyrdycz A, Laffargue P and Hildebrand HF: In vitro studies of human and rat osteoclast activity on hydroxyapatite, beta-tricalcium phosphate, calcium carbonate. Biomol Eng. 19:143–152. 2002. View Article : Google Scholar : PubMed/NCBI

51 

Maeda K, Kobayashi Y, Udagawa N, Uehara S, Ishihara A, Mizoguchi T, Kikuchi Y, Takada I, Kato S, Kani S, et al: Wnt5a-Ror2 signaling between osteoblast-lineage cells and osteoclast precursors enhances osteoclastogenesis. Nat Med. 18:405–412. 2012. View Article : Google Scholar : PubMed/NCBI

52 

Zhang X, Li Y, Chen YE, Chen J and Ma PX: Cell-free 3D scaffold with two-stage delivery of miRNA-26a to regenerate critical-sized bone defects. Nat Commun. 7:103762016. View Article : Google Scholar : PubMed/NCBI

53 

Sun LY, Wu L, Bao CY, Fu CH, Wang XL, Yao JF, Zhang XD and van Blitterswijk CA: Gene expressions of collagen type I, ALP and BMP-4 in osteoinductive BCP implants show similar pattern to that of natural healing bones. Mater Sci Eng C. 29:1829–1834. 2009. View Article : Google Scholar

54 

Wang J, Chen Y, Zhu X, Yuan T, Tan Y, Fan Y and Zhang X: Effect of phase composition on protein adsorption and osteoinduction of porous calcium phosphate ceramics in mice. J Biomed Mater Res A. 102:4234–4243. 2014.PubMed/NCBI

55 

Yi T, Jun CM, Kim SJ and Yun JH: Evaluation of in vivo osteogenic potential of bone morphogenetic Protein 2-Overexpressing human periodontal ligament stem cells combined with biphasic calcium phosphate block scaffolds in a Critical-Size bone defect model. Tissue Eng Part A. 22:501–512. 2016. View Article : Google Scholar : PubMed/NCBI

56 

Viti F, Landini M, Mezzelani A, Petecchia L, Milanesi L and Scaglione S: Osteogenic differentiation of MSC through calcium signaling activation: Transcriptomics and functional analysis. PLoS One. 11:pp. e01481732016, View Article : Google Scholar : PubMed/NCBI

57 

Tang Z, Tan Y, Ni Y, Wang J, Zhu X, Fan Y, Chen X, Yang X and Zhang X: Comparison of ectopic bone formation process induced by four calcium phosphate ceramics in mice. Mater Sci Eng C Mater Biol Appl. 70:1000–1010. 2017. View Article : Google Scholar

58 

González-Vázquez A, Planell JA and Engel E: Extracellular calcium and CaSR drive osteoinduction in mesenchymal stromal cells. Acta Biomater. 10:2824–2833. 2014. View Article : Google Scholar : PubMed/NCBI

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September 2019
Volume 44 Issue 3

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Online ISSN:1791-244X

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Copy and paste a formatted citation
APA
Yuan, X., Han, L., Lin, H., Guo, Z., Huang, Y., Li, S. ... Long, J. (2019). The role of antimiR-26a-5p/biphasic calcium phosphate in repairing rat femoral defects. International Journal of Molecular Medicine, 44, 857-870. https://doi.org/10.3892/ijmm.2019.4249
MLA
Yuan, X., Han, L., Lin, H., Guo, Z., Huang, Y., Li, S., Long, T., Tang, W., Tian, W., Long, J."The role of antimiR-26a-5p/biphasic calcium phosphate in repairing rat femoral defects". International Journal of Molecular Medicine 44.3 (2019): 857-870.
Chicago
Yuan, X., Han, L., Lin, H., Guo, Z., Huang, Y., Li, S., Long, T., Tang, W., Tian, W., Long, J."The role of antimiR-26a-5p/biphasic calcium phosphate in repairing rat femoral defects". International Journal of Molecular Medicine 44, no. 3 (2019): 857-870. https://doi.org/10.3892/ijmm.2019.4249