Low-intensity photobiomodulation at 632.8 nm increases tgfβ3, col2a1, and sox9 gene expression in rat bone marrow mesenchymal stem cells in vitro


The high incidence of cartilage destructions, as well as the social and economic importance of this pathology attracted great interest to the problem. At the present time, some data are available about the 632.8 nm low-intensity laser photobiomodulation positive effect on the cartilage tissue proliferation. The effect of this wavelength laser irradiation on the mesenchimal stem cell (MSC) differentiation in the chondrogenic direction was studied. The main aim of this work was to assess the low-intensity photobiomodulation effect on chondrogenesis. In this experiment, the cell model was used to compare the photobiomodulation and cytokine Tgfβ3 (transforming growth factor β 3) effects. Bone marrow MSCs were isolated from Wistar rats and cultured for the third passage. Chondrogenic effects of low-intensity He-Ne laser photobiomodulation and cytokine Tgfβ3 (10 ng/μL) were analyzed and compared after 21 days. The radiation source was the standard LGN-208 helium-neon (He–Ne) laser (632.8 nm, 1.7 mWt). Irradiation was performed cyclically for 15 min with 45-min pauses. The increase of the responsible for chondrogenesis (col2a1, tgfβ3, and sox9) main gene expression under the photobiomodulation at 632.8 nm was evaluated in comparison with Tgfβ3 effect. The tgfβ3, col2a1, and sox9 gene expression increase was obtained in two experimental groups: using the laser photobiomodulation and cytokine Tgfβ3 effect. Gene expression levels of tgfβ3, col2a1, and sox9 were measured using real-time polymerase chain reaction (RT-PCR) according to the -ΔΔCt method. It was found that the responsible for chondrogenesis genes expression (tgfβ3, col2a1, sox9) increased under the action of specific laser photobiomodulation during the observation period (from 0 to 21 days). The chondrogenic differentiation effect under the laser irradiation is less significant than Tgfβ3 cytokine effect.

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Mesenchymal stem cells




Transforming growth factor beta-3 protein


Polymerase chain reaction


Reverse transcription polymerase chain reaction


  1. 1.

    Bozhokin MS, Bozhkova SA, Netylko GI (2016) Possibilities of current cellular technologies for articular cartilage repair (Analytical review). Traumatol Orthop Russia 22(3):122–134. https://doi.org/10.21823/2311-2905-2016-22-3-122-134

    Article  Google Scholar 

  2. 2.

    Vassão PG, de Souza MC, Silva BA, Junqueira RG, de Camargo MR, Dourado VZ, Tucci HT, Renno AC (2020) Photobiomodulation via a cluster device associated with a physical exercise program in the level of pain and muscle strength in middle-aged and older women with knee osteoarthritis: a randomized placebo-controlled trial. Lasers Med Sci 35(1):139–148. https://doi.org/10.1007/s10103-019-02807-3

    Article  PubMed  Google Scholar 

  3. 3.

    Medvedeva EV, Grebenik EA, Gornostaeva SN, Telpuhov VI, Lychagin AV, Timashev PS, Chagin AS (2018) Repair of damaged articular cartilage: current approaches and future directions. Review Int J Mol Sci 19(8):2366. https://doi.org/10.3390/ijms19082366

    CAS  Article  Google Scholar 

  4. 4.

    Murray CJL, Vos T, Lozano R, Naghavi M, Flaxman AD, Michaud C, Ezzati M, Shibuya K, Salomon JA, Abdalla S et al (2012) Disability-adjusted life years (DALYs) for 291 diseases and injuries in 21 regions, 1990–2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet 380:2197–2223. https://doi.org/10.1016/S0140-6736(12)61689-4

    Article  PubMed  Google Scholar 

  5. 5.

    Bjordal JM, Couppe С, Chow RT, Ljunggren EA (2003) A systematic review of low level laser therapy with location-specific doses for pain from chronic joint disorders. Aust J Physiother 49(2):107–122. https://doi.org/10.1016/s0004-9514(14)60127-6

    Article  PubMed  Google Scholar 

  6. 6.

    Sobol E, Shekhter A, Guller A, Baum O, Baskov A (2011) Laser-induced regeneration of cartilage. J Biomed Opt 16(8):080902(2011). https://doi.org/10.1117/1.3614565

    Article  Google Scholar 

  7. 7.

    Bublitz C, Medalha C, Oliveira P, Assis L, Milares LP, Fernandes KR, Tim CR, Vasilceac FA, Mattiello SM, Renno ACM (2014) Low-level laser therapy prevents degenerative morphological changes in an experimental model of anterior cruciate ligament transection in rats. Lasers Med Sci 29(5):1669–1678. https://doi.org/10.1007/s10103-014-1546-z

    Article  PubMed  Google Scholar 

  8. 8.

    Pfander D, Jörgensen B, Rohde E, Bindig U, Müller G, Eric SE (2006) The influence of laser irradiation of low-power density on an experimental cartilage damage in rabbit knee-joints: an in vivo investigation considering macroscopic, histological and immunohistochemical changes. Biomed Tech (Berl) 51(3):131–138. https://doi.org/10.1515/BMT.2006.022

    Article  Google Scholar 

  9. 9.

    de Faria Coelho C, Leal-Junior ECP, Biasotto-Gonzalez DA, Bley AS, de Tarso de Camillo de Carvalho P, Politti F, de Oliveira Gonzalez T, de Oliveira AR, Frigero M, Garcia MBS, Dibai-Filho AV, de Paula Gomes CAF (2014) Effectiveness of phototherapy incorporated into an exercise program for osteoarthritis of the knee: study protocol for a randomized controlled trial. Trials 15:221. https://doi.org/10.1186/1745-6215-15-221

    Article  Google Scholar 

  10. 10.

    de Paula Gomes CAF, Leal-Junior ECP, Dibai-Filho AV, de Oliveira AR, Bley AS, Biasotto-Gonzalez DA, de Tarso Camillo de Carvalho P (2018) Incorporation of photobiomodulation therapy into a therapeutic exercise program for knee osteoarthritis: a placebo-controlled, randomized, clinical. Lasers Surg Med 50(8):819–828. https://doi.org/10.1002/lsm.22939

    Article  PubMed  Google Scholar 

  11. 11.

    Wei X, Heilig J, Paulsson M, Zaucke F (2015) Collagen II regulates chondrocyte integrin expression profile and differentiation. Connect Tissue Res 56(4):307–314. https://doi.org/10.3109/03008207.2015.1026965

    CAS  Article  Google Scholar 

  12. 12.

    Yang Q, Teng B, Wang L, Li K, Xu C, Ma X, Zhang Y, Kong D, Wang L, Zhao Y (2017) Silk fibroin/cartilage extracellular matrix scaffolds with sequential delivery of TGF-β3 for chondrogenic differentiation of adipose-derived stem cells. Int J Nanomedicine 12:6721–6733. https://doi.org/10.2147/IJN.S141888

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Lefebvre V, Angelozzi M, Haseeb A (2019) SOX9 in Cartilage Development and Disease. Review Curr Opin Cell Biol 61:39–47. https://doi.org/10.1016/j.ceb.2019.07.008

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Hata A, Chen Y (2016) TGF-β signaling from receptors to smads. Perspect Biol 8(9):a022061. https://doi.org/10.1101/cshperspect.a022061

    CAS  Article  Google Scholar 

  15. 15.

    Nazempour A, Quisenberry CR, Van Wie BJ, Abu-Lail NI (2016) Nanomechanics of engineered articular cartilage: synergistic influences of transforming growth factor-β3 and oscillating pressure. J Nanosci Nanotechnol 16(3):3136–3145. https://doi.org/10.1166/jnn.2016.12564

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Labutin D, Bozhokin MS (2016) Optimization of the method for mesenchymal stem cell isolation from rat bone marrow. Conference: 2016 Conference for Young Scientists of the Northwestern Federal District, St. Petersburg, Russian Federation. https://doi.org/10.13140/RG.2.1.2723.8807

  17. 17.

    Goessler UR, Bugert P, Bieback K, Deml M, Sadick H, Hormann K, Riedel F (2005) In-vitro analysis of the expression of TGF beta-superfamily-members during chondrogenic differentiation of mesenchymal stem cells and chondrocytes during dedifferentiation in cell culture. Cell Mol Biol Lett 10(2):345–362

    CAS  PubMed  Google Scholar 

  18. 18.

    Farivar S, Ramezankhani R, Mohajerani E, Ghazimoradi MH, Shiari R (2019) Gene expression analysis of chondrogenic markers in hair follicle dermal papillae cells under the effect of laser photobiomodulation and the synovial fluid. J Lasers Med Sci 10(3):171–178. https://doi.org/10.15171/jlms.2019.27

    Article  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Holden PK, Li C, Da Costa V, Sun CH, Bryant SV, Gardiner DM, Wong BJ (2009) The effects of laser irradiation of cartilage on chondrocyte gene expression and the collagen matrix. Lasers Surg Med 41(7):487–491. https://doi.org/10.1002/lsm.20795

    Article  PubMed  PubMed Central  Google Scholar 

  20. 20.

    Milares LP, Assis L, Siqueira A, Claudino V, Domingos H, Almeida T, Tim C, Renno AC (2016) Effectiveness of an aquatic exercise program and low-level laser therapy on articular cartilage in an experimental model of osteoarthritis in rats. Connect Tissue Res 57(5):398–407. https://doi.org/10.1080/03008207.2016.1193174

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Kushibiki T, Tajiri T, Ninomiya Y, Awazu K (2010) Chondrogenic mRNA expression in prechondrogenic cells after blue laser irradiation. J Photochem Photobiol B98(3):211–215. https://doi.org/10.1016/j.jphotobiol.2010.01.008

    CAS  Article  Google Scholar 

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This work was supported by the Ministry of Science and Higher Education within the State assignment no. 0103-2019-0012.

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Correspondence to M. S. Bozhokin.

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Bozhokin, M.S., Vcherashnii, D.B., Yastrebov, S.G. et al. Low-intensity photobiomodulation at 632.8 nm increases tgfβ3, col2a1, and sox9 gene expression in rat bone marrow mesenchymal stem cells in vitro. Lasers Med Sci (2021). https://doi.org/10.1007/s10103-021-03279-0

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  • Low-intensity laser radiation
  • Chondrogenesis
  • col2a1
  • tgfβ3
  • sox9 gene expression