Skip to main content

Advertisement

Springer Nature Link
Log in

Wound-healing effects of 635-nm low-level laser therapy on primary human vocal fold epithelial cells: an in vitro study

  • Original Article
  • Published:
Lasers in Medical Science Aims and scope Submit manuscript

Abstract

Low-level laser therapy (LLLT) has been promoted for its beneficial effects on tissue healing and pain relief for skin and oral applications. However, there is no corresponding literature reporting on vocal fold wound healing. Our purpose was to assess the potential wound-healing effects of LLLT on primary human vocal fold epithelial cells (VFECs). In this study, normal vocal fold tissue was obtained from a 58-year-old male patient who was diagnosed with postcricoid carcinoma without involvement of the vocal folds and underwent total laryngectomy. Primary VFECs were then cultured. Cells were irradiated at a wavelength of 635 nm with fluences of 1, 4, 8, 12, 16, and 20 J/cm2 (50 mW/cm2), which correspond to irradiation times of 20, 80, 160, 240, 320, and 400 s, respectively. Cell viability of VFECs in response to varying doses of LLLT was investigated by the Cell Counting Kit-8 (CCK-8) method. The most effective irradiation dose was selected to evaluate the cell migration capacity by using the scratch wound-healing assay. Real-time polymerase chain reaction (RT-PCR) was used to detect the gene expression of TGF-β1, TGF-β3, EGF, IL-6, and IL-10. Irradiation with doses of 8 J/cm2 resulted in 4% increases in cell proliferation differing significantly from the control group (p < 0.05). With subsequent doses at 48 and 72 h after irradiation, the differences between the experimental and the control groups became greater, up to 9.8% (p < 0.001) and 19.5% (p < 0.001), respectively. It also increased cell migration and the expression of some genes, such as EGF, TGF-β1, TGF-β3, and IL-10, involved in the tissue healing process. This study concludes that LLLT at the preset parameters was capable of stimulating the proliferation and migration of human vocal fold epithelial cells in culture as well as increase the expression of some genes involved in tissue healing process. Additionally, successive laser treatments at 24 h intervals have an additive beneficial effect on the healing of injured tissues.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /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.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Friedrich G, Dikkers FG, Arens C, Remacle M, Hess M, Giovanni A, Duflo S, Hantzakos A, Bachy V, Gugatschka M, European Laryngological Society. Phonosurgery Committe (2013) Vocal fold scars: current concepts and future directions. Consensus report of the Phonosurgery Committee of the European Laryngological Society. Eur Arch Otorhinolaryngol 270:2491–2507

    Article  PubMed  CAS  Google Scholar 

  2. Abramoff MM, Lopes NN, Lopes LA, Dib LL, Guilherme A, Caran EM, Barreto AD, Lee ML, Petrilli AS (2008) Low-level laser therapy in the prevention and treatment of chemotherapy-induced oral mucositis in young patients. Photomed Laser Surg 26:393–400

    Article  PubMed  CAS  Google Scholar 

  3. Sugrue ME, Carolan J, Leen EJ, Feeley TM, Moore DJ, Shanik GD (1990) The use of infrared laser therapy in the treatment of venous ulceration. Ann Vasc Surg 4:179–181

    Article  PubMed  CAS  Google Scholar 

  4. Schindl A, Schindl M, Schindl L (1997) Successful treatment of a persistent radiation ulcer by low power laser therapy. J Am Acad Dermatol 37:646–648

    Article  PubMed  CAS  Google Scholar 

  5. Häkkinen L, Uitto VJ, Larjava H (2010) Cell biology of gingival wound healing. Periodontol 24:127–152

    Article  Google Scholar 

  6. Kreisler M, Christoffers AB, Al-Haj H, Willershausen B, d’Hoedt B (2002) Low level 809-nm diode laser-induced in vitro stimulation of the proliferation of human gingival fibroblasts. Lasers Surg Med 30:365–369

    Article  PubMed  Google Scholar 

  7. Posten W, Wrone DA, Dover JS, Arndt KA, Silapunt S, Alam M (2005) Low-level laser therapy for wound healing: mechanism and efficacy. Dermatol Surg 31:334–340

    Article  PubMed  CAS  Google Scholar 

  8. Leydon C, Imaizumi M, Bartlett RS, Wang SF, Thibeault SL (2014) Epithelial cells are active participants in vocal fold wound healing: an in vivo animal model of injury. PLoS One 9:e115389

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  9. Mizuta M, Kurita T, Kimball EE, Rousseau B (2017) Structurally and functionally characterized in vitro model of rabbit vocal fold epithelium. Tissue Cell 49:427–434

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Ren C, Mcgrath C, Jin L, Zhang C, Yang Y (2016) Effect of diode low-level lasers on fibroblasts derived from human periodontal tissue: a systematic review of in vitro studies. Lasers Med Sci 31:1–18

    Article  Google Scholar 

  11. Mester E, Mester AF, Mester A (1985) The biomedical effects of laser application. Lasers Surg Med 5:31–39

    Article  PubMed  CAS  Google Scholar 

  12. Marques MM, Pereira AN, Fujihara NA, Nogueira FN, Eduardo CP (2004) Effect of low-power laser irradiation on protein synthesis and ultrastructure of human gingival fibroblasts. Lasers Surg Med 34:260–265

    Article  PubMed  Google Scholar 

  13. Ayuk SM, Houreld NN, Abrahamse H (2018) Effect of 660 nm visible red light on cell proliferation and viability in diabetic models in vitro under stressed conditions. Lasers Med Sci 33:1085–1093

    Article  PubMed  CAS  Google Scholar 

  14. Alghamdi KM, Kumar A, Moussa NA (2012) Low-level laser therapy: a useful technique for enhancing the proliferation of various cultured cells. Lasers Med Sci 27:237–249

    Article  PubMed  Google Scholar 

  15. Fekrazad R, Asefi S, Allahdadi M, Kalhori KA (2016) Effect of photobiomodulation on mesenchymal stem cells. Photomed Laser Surg 34:533–542

    Article  PubMed  Google Scholar 

  16. Hawkins DH, Abrahamse H (2006) The role of laser fluence in cell viability, proliferation, and membrane integrity of wounded human skin fibroblasts following helium-neon laser irradiation. Lasers Surg Med 38:74–83

    Article  PubMed  Google Scholar 

  17. Wilson SE, Chen L, Mohan RR, Liang Q, Liu J (1999) Expression of HGF, KGF, EGF and receptor messenger RNAs following corneal epithelial wounding. Exp Eye Res 68:377–397

    Article  PubMed  CAS  Google Scholar 

  18. Barrientos S, Stojadinovic O, Golinko MS, Brem H, Tomic-Canic M (2008) Growth factors and cytokines in wound healing. Wound Repair Regen 16:585–601

    Article  PubMed  Google Scholar 

  19. Puddicombe SM, Polosa R, Richter A, Krishna MT, Howarth PH, Holgate ST, Davies DE (2000) Involvement of the epidermal growth factor receptor in epithelial repair in asthma. FASEB J 14:1362–1374

    Article  PubMed  CAS  Google Scholar 

  20. Boxall C, Holgate ST, Davies DE (2006) The contribution of transforming growth factor-beta and epidermal growth factor signalling to airway remodelling in chronic asthma. Eur Respir J 27:208–229

    Article  PubMed  CAS  Google Scholar 

  21. Znalesniak EB, Hoffmann W (2010) Modulation of cell-cell contacts during intestinal restitution in vitro and effects of epidermal growth factor (EGF). Cell Physiol Biochem 25:533–542

    Article  PubMed  CAS  Google Scholar 

  22. Ferguson MW, O'Kane S (2004) Scar-free healing: from embryonic mechanisms to adult therapeutic intervention. Philos Trans R Soc Lond Ser B Biol Sci 359:839–850

    Article  CAS  Google Scholar 

  23. Whitby DJ, Ferguson MW (1991) Immunohistochemical localization of growth factors in fetal wound healing. Dev Biol 147:207–215

    Article  PubMed  CAS  Google Scholar 

  24. Hamblin MR, Demidova TN (2006) Mechanisms of low level light therapy. Proc SPIE 6140:1–12

    Google Scholar 

  25. Toyokawa H, Matsui Y, Uhara J, Tsuchiya H, Teshima S, Nakanishi H, Kwon AH, Azuma Y, Nagaoka T, Ogawa T, Kamiyama Y (2003) Promotive effects of far-infrared ray on full-thickness skin wound healing in rats. Exp Biol Med (Maywood) 228:724–729

    Article  CAS  Google Scholar 

  26. Arany PR, Nayak RS, Hallikerimath S, Limaye AM, Kale AD, Kondaiah P (2007) Activation of latent TGF-beta1 by low-power laser in vitro correlates with increased TGF-beta1 levels in laser-enhanced oral wound healing. Wound Repair Regen 15:866–874

    Article  PubMed  Google Scholar 

  27. Chang Z, Kishimoto Y, Hasan A, Welham NV (2014) TGF-β3 modulates the inflammatory environment and reduces scar formation following vocal fold mucosal injury in rats. Dis Model Mech 7:83–91

    Article  PubMed  CAS  Google Scholar 

  28. Lin TW, Cardenas L, Glaser DL, Soslowsky LJ (2006) Tendon healing in interleukin-4 and interleukin-6 knockout mice. J Biomech 39:61–69

    Article  PubMed  Google Scholar 

  29. Skutek M, van Griensven M, Zeichen J, Brauer N, Bosch U (2001) Cyclic mechanical stretching enhances secretion of interleukin 6 in human tendon fibroblasts. Knee Surg Sports Traumatol Arthrosc 9:322–326

    Article  PubMed  CAS  Google Scholar 

  30. Ricchetti ET, Reddy SC, Ansorge HL, Zgonis MH, Van Kleunen JP, Liechty KW, Soslowsky LJ, Beredjiklian PK (2008) Effect of interleukin-10 overexpression on the properties of healing tendon in a murine patellar tendon model. J Hand Surg Am 33:1843–1852

    Article  PubMed  Google Scholar 

  31. Alam R, Kumar D, Anderson-Walters D, Forsythe PA (1994) Macrophage inflammatory protein-1 alpha and monocyte chemoattractant peptide-1 elicit immediate and late cutaneous reactions and activate murine mast cells in vivo. J Immunol 152:1298–1303

    PubMed  CAS  Google Scholar 

  32. Lee SY, Park KH, Choi JW, Kwon JK, Lee DR, Shin MS, Lee JS, You CE, Park MY (2007) A prospective, randomized, placebo-controlled, double-blinded, and split-face clinical study on LED phototherapy for skin rejuvenation: clinical, profilometric, histologic, ultrastructural, and biochemical evaluations and comparison of three different trea. J Photochem Photobiol B 88:51–67

    Article  PubMed  CAS  Google Scholar 

  33. Torres-Silva R, Lopes-Martins RA, Bjordal JM, Frigo L, Rahouadj R, Arnold G, Leal-Junior EC, Magdalou J, Pallotta R, Marcos RL (2015) The low level laser therapy (LLLT) operating in 660 nm reduce gene expression of inflammatory mediators in the experimental model of collagenase-induced rat tendinitis. Lasers Med Sci 30:1985–1990

    Article  PubMed  Google Scholar 

  34. Schindl A, Schindl M, Pernerstorfer-Schon H, Schindl L (2000) Low-intensity laser therapy: a review. J Investig Med 48:312–326

    PubMed  CAS  Google Scholar 

  35. Schindl A, Schindl M, Schon H, Knobler R, Havelec L, Schindl L (1998) Low-intensity laser irradiation improves skin circulation in patients with diabetic microangiopathy. Diabetes Care 21:580–584

    Article  PubMed  CAS  Google Scholar 

  36. Karu TI, Pyatibrat LV, Kalendo GS (2004) Photobiological modulation of cell attachment via cytochrome c oxidase. Photochem Photobiol Sci 3:211–216

    Article  PubMed  CAS  Google Scholar 

  37. Avci P, Gupta A, Sadasivam M, Vecchio D, Pam Z, Pam N, Hamblin MR (2013) Low-level laser (light) therapy (LLLT) in skin: stimulating, healing, restoring. Semin Cutan Med Surg 32:41–52

    PubMed  PubMed Central  Google Scholar 

Download references

Role of funding source

The work is supported by the National Natural Science Foundation of China (number: 81329001).

Author information

Authors and Affiliations

  1. The Department of Otolaryngology—Head and Neck Surgery, Eye, Ear, Nose, and Throat Hospital, Fudan University, 83 Fenyang Road, Shanghai, 200030, China

    Zhewei Lou, Chi Zhang, Ting Gong, Chao Xue & Jack J. Jiang

  2. The Department of Surgery, Division of Otolaryngology—Head and Neck Surgery, University of Wisconsin-Madison School of Medicine and Public Health, Madison, WI, USA

    Austin Scholp

Authors
  1. Zhewei Lou
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Chi Zhang
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Ting Gong
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Chao Xue
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Austin Scholp
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Jack J. Jiang
    View author publications

    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence to Jack J. Jiang.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

Ethical approval was obtained from the ethics committee of the Eye, Ear, Nose, and Throat Hospital of Fudan University (2017061).

Informed consent

Informed consent was obtained from all individual participants included in the study.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lou, Z., Zhang, C., Gong, T. et al. Wound-healing effects of 635-nm low-level laser therapy on primary human vocal fold epithelial cells: an in vitro study. Lasers Med Sci 34, 547–554 (2019). https://doi.org/10.1007/s10103-018-2628-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10103-018-2628-0

Keywords