Improvement in viability and mineralization of osteoporotic bone marrow mesenchymal stem cell through combined application of photobiomodulation therapy and oxytocin

Abstract

The probable positive effects of photobiomodulation therapy (PBMT) and oxytocin (OT) treatments together or alone were evaluated on cell viability along with the changes in the gene expression of Osteocalcin (OC), Osteoprotegerin (OPG), and Runt-related transcription factor 2 (Runx2) levels of sham (healthy)-Bone marrow mesenchymal stem cell(BMMSC) and ovariectomy-induced osteoporosis (OVX)-BMMSC. BMMSC was harvested from healthy and OVX rats and was cultured in osteogenic induction medium (OIM). There were five groups of BMMSCs: (1) sham -BMMSCs; (2) control -OVX-BMMSCs; (3) OT-treated-OVX-BMMSCs; (4) PBMT–treated-OVX-BMMSCs, and (5) OT + PBMT-OVX-BMMSCs. In all 5 groups, BMMSC viability and proliferation as well as gene expression of OC, OPG, and RUNX2 were evaluated. PBMT and PBMT + OT treatments showed a promising effect on the increased viability of OVX-BMMSC (ANOVA test; LSD test, p = 0.01, p = 0.002). The results of gene expression analysis revealed that the sham- BMMSCs responded optimally to OT treatment. It was also found that OVX-BMMSCs responded optimally to PBMT + OT and PBMT treatments at early and middle stages of osteogenic induction process. Nevertheless, they responded optimally to PBMT + OT and OT especially at the late stage of osteogenic induction process. PBMT and PBMT + OT treatments significantly increased viability of OVX-BMMSC in OIM in vitro. Both PBMT and PBMT + OT treatments could promote mineralization of OVX-BMMSC in the culture medium at early and middle stages of osteogenic induction process. Both OT and PBMT + OT treatments could promote mineralization of OVX-BMMSC in vitro at late stages of osteogenic induction process.

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References

  1. 1.

    Rucci N (2008) Molecular biology of bone remodelling. Clin Cases Miner Bone Metab 5:49

    PubMed  PubMed Central  Google Scholar 

  2. 2.

    Sözen T, Özışık L, Başaran NÇ (2017) An overview and management of osteoporosis. Eur J Rheumatol 4:46

    PubMed  Article  Google Scholar 

  3. 3.

    Bielby R, Jones E, McGonagle D (2007) The role of mesenchymal stem cells in maintenance and repair of bone. Injury. 38:S26–S32

    PubMed  Article  Google Scholar 

  4. 4.

    Tan J, Xu X, Tong Z, Yu Q, Lin Y, Kuang W (2015) Decreased osteogenesis of adult mesenchymal stem cells by reactive oxygen species under cyclic stretch: a possible mechanism of age related osteoporosis. Bone Res 3:15003

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  5. 5.

    Rodríguez JP, Garat S, Gajardo H, Pino AM, Seitz G (1999) Abnormal osteogenesis in osteoporotic patients is reflected by altered mesenchymal stem cells dynamics. J Cell Biochem 75:414–423

    PubMed  Article  Google Scholar 

  6. 6.

    Kiernan J, Davies JE, Stanford WL (2017) Concise review: musculoskeletal stem cells to treat age-related osteoporosis. Stem Cells Transl Med 6:1930–1939

    PubMed  PubMed Central  Article  Google Scholar 

  7. 7.

    Bonyadi M, Waldman SD, Liu D, Aubin JE, Grynpas MD, Stanford WL (2003) Mesenchymal progenitor self-renewal deficiency leads to age-dependent osteoporosis in Sca-1/Ly-6A null mice. Proc Natl Acad Sci 100:5840–5845

    CAS  PubMed  Article  Google Scholar 

  8. 8.

    Pignolo RJ, Suda RK, McMillan EA, Shen J, Lee SH, Choi Y, Wright AC, Johnson FB (2008) Defects in telomere maintenance molecules impair osteoblast differentiation and promote osteoporosis. Aging Cell 7:23–31

    CAS  PubMed  Article  Google Scholar 

  9. 9.

    Tewari D, Khan MP, Sagar N, China SP, Singh AK, Kheruka SC, Barai S, Tewari MC, Nagar GK, Vishwakarma AL (2015) Ovariectomized rats with established osteopenia have diminished mesenchymal stem cells in the bone marrow and impaired homing, osteoinduction and bone regeneration at the fracture site. Stem Cell Rev Rep 11:309–321

    CAS  PubMed  Article  Google Scholar 

  10. 10.

    An JH, Park H, Song JA, Ki KH, Yang J-Y, Choi HJ, Cho SW, Kim SW, Kim SY, Yoo JJ (2013) Transplantation of human umbilical cord blood-derived mesenchymal stem cells or their conditioned medium prevents bone loss in ovariectomized nude mice. Tissue Eng A 19:685–696

    CAS  Article  Google Scholar 

  11. 11.

    Lindtner RA, Tiaden AN, Genelin K, Ebner HL, Manzl C, Klawitter M, Sitte I, von Rechenberg B, Blauth M, Richards PJ (2014) Osteoanabolic effect of alendronate and zoledronate on bone marrow stromal cells (BMSCs) isolated from aged female osteoporotic patients and its implications for their mode of action in the treatment of age-related bone loss. Osteoporos Int 25:1151–1161

    CAS  PubMed  Article  Google Scholar 

  12. 12.

    Nakamura A, Dohi Y, Akahane M, Ohgushi H, Nakajima H, Funaoka H, Takakura Y (2009) Osteocalcin secretion as an early marker of in vitro osteogenic differentiation of rat mesenchymal stem cells. Tissue Eng Part C Methods 15:169–180

    CAS  PubMed  Article  Google Scholar 

  13. 13.

    Boyce BF, Xing L (2008) Functions of RANKL/RANK/OPG in bone modeling and remodeling. Arch Biochem Biophys 473:139–146

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  14. 14.

    Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G (1997) Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89:747–754

    CAS  PubMed  Article  Google Scholar 

  15. 15.

    Khalid O, Baniwal SK, Purcell DJ, Leclerc N, Gabet Y, Stallcup MR, Coetzee GA, Frenkel B (2008) Modulation of Runx2 activity by estrogen receptor-alpha: implications for osteoporosis and breast cancer. Endocrinology. 149:5984–5995

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  16. 16.

    Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, Sato M, Okamoto R, Kitamura Y, Yoshiki S, Kishimoto T (1997) Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. 89:755–764

    CAS  PubMed  Article  Google Scholar 

  17. 17.

    Salingcarnboriboon R, Tsuji K, Komori T, Nakashima K, Ezura Y, Noda M (2006) Runx2 is a target of mechanical unloading to alter osteoblastic activity and bone formation in vivo. Endocrinology. 147:2296–2305

    CAS  PubMed  Article  Google Scholar 

  18. 18.

    Verma SK, Maheshwari S, Singh RK, Chaudhari PK (2012) Laser in dentistry: an innovative tool in modern dental practice. Natl J Maxillofac Surg 3:124

    PubMed  PubMed Central  Article  Google Scholar 

  19. 19.

    de Freitas LF, Hamblin MR (2016) Proposed mechanisms of photobiomodulation or low-level light therapy. IEEE J Sel Top Quantum Electron:22

  20. 20.

    Wang X, Tian F, Soni SS, Gonzalez-Lima F, Liu H (2016) Interplay between up-regulation of cytochrome-c-oxidase and hemoglobin oxygenation induced by near-infrared laser. Sci Rep 6:30540

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  21. 21.

    Wang X, Tian F, Reddy DD, Nalawade SS, Barrett DW, Gonzalez-Lima F, Liu H (2017) Up-regulation of cerebral cytochrome-c-oxidase and hemodynamics by transcranial infrared laser stimulation: a broadband near-infrared spectroscopy study. J Cereb Blood Flow Metab 37:3789–3802

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  22. 22.

    Fallahnezhad S, Amini A, Hajihossainlou B, Chien S, Dadras S, Rezaei F, Bayat M (2018) Combined effects of photobiomodulation and alendronate on viability of osteoporotic bone marrow-derived mesenchymal stem cells. J Photochem Photobiol B Biol 182:77–84

    CAS  Article  Google Scholar 

  23. 23.

    Fallahnezhad S, Piryaei A, Tabeie F, Nazarian H, Darbandi H, Amini A, Mostafavinia A, Ghorishi SK, Jalalifirouzkouhi A, Bayat M (2016) Low-level laser therapy with helium–neon laser improved viability of osteoporotic bone marrow-derived mesenchymal stem cells from ovariectomy-induced osteoporotic rats. J Biomed Opt 21:098002

    Article  Google Scholar 

  24. 24.

    Fallahnezhad S, Piryaei A, Darbandi H, Amini A, Ghoreishi SK, Jalalifirouzkouhi R, Bayat M (2018) Effect of low-level laser therapy and oxytocin on osteoporotic bone marrow-derived mesenchymal stem cells. J Cell Biochem 119:983–997

    CAS  PubMed  Article  Google Scholar 

  25. 25.

    Mostafavinia A, Dehdehi L, Ghoreishi SK, Hajihossainlou B, Bayat M (2017) Effect of in vivo low-level laser therapy on bone marrow-derived mesenchymal stem cells in ovariectomy-induced osteoporosis of rats. J Photochem Photobiol B Biol 175:29–36

    CAS  Article  Google Scholar 

  26. 26.

    Fridoni M, Farahani RM, Nejati H, Salimi M, Gharavi SM, Bayat M, Amini A, Torkman G, Bayat S (2015) Evaluation of the effects of LLLT on biomechanical properties of tibial diaphysis in two rat models of experimental osteoporosis by a three point bending test. Lasers Med Sci 30:1117–1125

    PubMed  Article  Google Scholar 

  27. 27.

    Mohsenifar Z, Fridoni M, Ghatrehsamani M, Abdollahifar M-A, Abbaszadeh H, Mostafavinia A, Fallahnezhad S, Asghari M, Bayat S, Bayat M (2016) Evaluation of the effects of pulsed wave LLLT on tibial diaphysis in two rat models of experimental osteoporosis, as examined by stereological and real-time PCR gene expression analyses. Lasers Med Sci 31:721–732

    PubMed  Article  Google Scholar 

  28. 28.

    Elabd C, Basillais A, Beaupied H, Breuil V, Wagner N, Scheideler M, Zaragosi LE, Massiéra F, Lemichez E, Trajanoski Z (2008) Oxytocin controls differentiation of human mesenchymal stem cells and reverses osteoporosis. Stem Cells 26:2399–2407

    CAS  PubMed  Article  Google Scholar 

  29. 29.

    Breuil V, Panaia-Ferrari P, Fontas E, Roux C, Kolta S, Eastell R, Ben Yahia H, Faure S, Gossiel F, Benhamou C-L (2014) Oxytocin, a new determinant of bone mineral density in post-menopausal women: analysis of the OPUS cohort. J Clin Endocrinol Metab 99:E634–E641

    CAS  PubMed  Article  Google Scholar 

  30. 30.

    Beranger GE, Djedaini M, Battaglia S, Roux CH, Scheideler M, Heymann D, Amri E-Z, Pisani DF (2015) Oxytocin reverses osteoporosis in a sex-dependent manner. Front Endocrinol 6:81

    Article  Google Scholar 

  31. 31.

    Wang Z, Goh J, De SD, Ge Z, Ouyang H, Chong JSW, Low SL, Lee EH (2006) Efficacy of bone marrow–derived stem cells in strengthening osteoporotic bone in a rabbit model. Tissue Eng 12:1753–1761

    CAS  PubMed  Article  Google Scholar 

  32. 32.

    Griffin MD, Ritter T, Mahon BP (2010) Immunological aspects of allogeneic mesenchymal stem cell therapies. Hum Gene Ther 21:1641–1655

    CAS  PubMed  Article  Google Scholar 

  33. 33.

    Prall WC, Haasters F, Heggebö J, Polzer H, Schwarz C, Gassner C, Grote S, Anz D, Jäger M, Mutschler W (2013) Mesenchymal stem cells from osteoporotic patients feature impaired signal transduction but sustained osteoinduction in response to BMP-2 stimulation. Biochem Biophys Res Commun 440:617–622

    CAS  PubMed  Article  Google Scholar 

  34. 34.

    Shen J, James AW, Zara JN, Asatrian G, Khadarian K, Zhang JB, Ho S, Kim HJ, Ting K, Soo C (2013) BMP2-induced inflammation can be suppressed by the osteoinductive growth factor NELL-1. Tissue Eng A 19:2390–2401

    CAS  Article  Google Scholar 

  35. 35.

    Saeed M, Sattari M, Yeganeh F, Shahnavaz S (2015) Cor-relation between gingival expression of STAT1 and chronic peri-odontitis. Int J Dent Oral Sci 2:158–162

    Google Scholar 

  36. 36.

    Wu JY, Wang YH, Wang GJ, Ho ML, Wang CZ, Yeh ML, Chen CH (2012) Low-power GaAlAs laser irradiation promotes the proliferation and osteogenic differentiation of stem cells via IGF1 and BMP2. PLoS One 7:e44027

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  37. 37.

    Baron W, Metz B, Bansal R, Hoekstra D, de Vries H (2000) PDGF and FGF-2 signaling in oligodendrocyte progenitor cells: regulation of proliferation and differentiation by multiple intracellular signaling pathways. Mol Cell Neurosci 15:314–329

    CAS  PubMed  Article  Google Scholar 

  38. 38.

    Chen W, Baylink DJ, Brier-Jones J, Neises A, Kiroyan JB, Rundle CH, Lau KH, Zhang XB (2015) PDGFB-based stem cell gene therapy increases bone strength in the mouse. Proc Natl Acad Sci U S A 112:E3893–E3900

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  39. 39.

    Baldo BA (2014) Side effects of cytokines approved for therapy. Drug Saf 37:921–943

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  40. 40.

    McLaughlin PJ, Cain JD, Titunick MB, Sassani JW, Zagon IS (2017) Topical naltrexone is a safe and effective alternative to standard treatment of diabetic wounds. Adv Wound Care 6:279–288

    Article  Google Scholar 

  41. 41.

    Hu C, Li L (2018) Preconditioning influences mesenchymal stem cell properties in vitro and in vivo. J Cell Mol Med 22:1428–1442

    PubMed  PubMed Central  Article  Google Scholar 

  42. 42.

    Noiseux N, Borie M, Desnoyers A, Menaouar A, Stevens LM, Mansour S, Danalache BA, Roy DC, Jankowski M, Gutkowska J (2012) Preconditioning of stem cells by oxytocin to improve their therapeutic potential. Endocrinology. 153:5361–5372

    CAS  PubMed  Article  Google Scholar 

  43. 43.

    Kim YS, Ahn Y, Kwon JS, Cho YK, Jeong MH, Cho JG, Park JC, Kang JC (2012) Priming of mesenchymal stem cells with oxytocin enhances the cardiac repair in ischemia/reperfusion injury. Cells Tissues Organs 195:428–442

    CAS  PubMed  Article  Google Scholar 

  44. 44.

    Zhang H, Li H (2018) Tricin enhances osteoblastogenesis through the regulation of Wnt/beta-catenin signaling in human mesenchymal stem cells. Mech Dev 152:38–43

    CAS  PubMed  Article  Google Scholar 

  45. 45.

    Liu TM, Lee EH (2013) Transcriptional regulatory cascades in Runx2-dependent bone development. Tissue Eng B Rev 19:254–263

    Article  CAS  Google Scholar 

  46. 46.

    Otto F, Thornell AP, Crompton T, Denzel A, Gilmour KC, Rosewell IR, Stamp GW, Beddington RS, Mundlos S, Olsen BR, Selby PB, Owen MJ (1997) Cbfa1, a candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development. Cell. 89:765–771

    CAS  PubMed  Article  Google Scholar 

  47. 47.

    Rocha Júnior AM, Vieira BJ, Andrade LCF d, Aarestrup FM (2007) Effects of low-level laser therapy on the progress of wound healing in humans: the contribution of in vitro and in vivo experimental studies. J Vasc Bras 6:257–265

    Article  Google Scholar 

  48. 48.

    Lev-Sagie A, Kopitman A, Brzezinski A (2017) Low-level laser therapy for the treatment of provoked vestibulodynia—a randomized, placebo-controlled pilot trial. J Sex Med 14:1403–1411

    PubMed  Article  Google Scholar 

  49. 49.

    Arunachalam LT, Sudhakar U, Janarthanam AS, Das NM (2014) Effect of low level laser therapy on revascularization of free gingival graft using ultrasound Doppler flowmetry. J Indian Soc Periodontol 18:403

    PubMed  PubMed Central  Article  Google Scholar 

  50. 50.

    Cassano P, Tran AP, Katnani H, Bleier BS, Hamblin MR, Yuan Y, Fang Q (2019) Selective photobiomodulation for emotion regulation: model-based dosimetry study. Neurophotonics. 6:015004

    CAS  PubMed  PubMed Central  Article  Google Scholar 

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Funding

This article was financially supported by the Research Department of the Shahid Beheshti University of Medical Sciences, Tehran, Iran (Grant no 1393-1-91-13237).

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Correspondence to Mohammad Bayat.

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Fallahnezhad, S., Jajarmi, V., Shahnavaz, S. et al. Improvement in viability and mineralization of osteoporotic bone marrow mesenchymal stem cell through combined application of photobiomodulation therapy and oxytocin. Lasers Med Sci 35, 557–566 (2020). https://doi.org/10.1007/s10103-019-02848-8

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Keywords

  • Bone marrow mesenchymal stem cells
  • Low-level laser therapy
  • Photobiomodulation therapy
  • Oxytocin
  • Ovariectomy
  • Post-menopausal osteoporosis
  • Osteocalcin
  • Osteoprotegerin
  • Runt-related transcription factor 2