Lasers in Medical Science

, Volume 30, Issue 2, pp 533–541 | Cite as

Vascular regeneration effect of adipose-derived stem cells with light-emitting diode phototherapy in ischemic tissue

  • In-Su Park
  • Arindam Mondal
  • Phil-Sang Chung
  • Jin Chul Ahn
Original Article


The objective of this study was to investigate the effects on the vascular regeneration of adipose-derived stem cells (ASCs) by using red light-emitting diode (LED) irradiation in ischemic hind limbs. Low-level light therapy (LLLT) has been shown to enhance proliferation and cytokine secretion of a number of cells. ASCs are an attractive cell source for vascular tissue engineering. This approach is hindered because transplanted ASCs decline rapidly in the recipient tissue. Ischemic hind limbs were treated with LLLT from an LED array (660 nm) at an irradiance of 50 mW/cm2 and a radiant exposure of 30 J/cm2. LLLT, ASC transplantation, and ASC transplantation with LLLT (ASC + LLLT) were applied to ischemic limbs, and cell survival and differentiation, and secretion of vascular endothelial growth factor and basic fibroblast growth factor of the ASCs were evaluated by immunostaining and Western blot analyses. Vascular regeneration was assessed by immunostaining and hematoxylin and eosin staining. In the ASC + LLLT group, the survival of ASCs was increased due to the decreased apoptosis of ASCs. The secretion of growth factors was stimulated in this group compared with ASCs alone. The ASC + LLLT group displayed improved treatment efficacy including neovascularization and tissue regeneration compared with ASCs alone. In particular, quantitative analysis of laser Doppler blood perfusion image ratio showed that blood perfusion was enhanced significantly (p < 0.05) by ASC + LLLT treatment. These data suggest that LLLT is an effective biostimulator of ASCs in vascular regeneration, which enhances the survival of ASCs and stimulates the secretion of growth factors in ischemic limbs.


Adipose-derived stromal cell Angiogenesis Low-level light Vascular endothelial growth factor Ischemic limbs 



This study was supported by a grant of the Ministry of Science, ICT and Future Planning funded by the Korea government (2012K1A4A3053142, NRF-2014R1A1A1038199).

Supplementary material

10103_2014_1699_MOESM1_ESM.tif (1.9 mb)
Figure 1 Immunofluorescence staining and flow cytometry analyses of hASCs. (A) hASCs (passage 4) were stained with CD29, CD90 and CD105 for mesenchymal stem cell identification, with KDR, CD31 and CD34 for endothelial lineage cell identification, and SMA for smooth muscle cell identification. Scale bar: 200 μm (B) Flow cytometry analysis; hASCs cultured for 1 days were stained for CD29, CD90, CD105, CD45, CD31, CD34 and KDR expression and analyzed by flow cytometry. (TIFF 1962 kb)
10103_2014_1699_Fig6_ESM.gif (98 kb)

(GIF 97 kb)

10103_2014_1699_MOESM2_ESM.tif (2.1 mb)
Table 1 List of antibodies for immunofluorescence staining. (TIFF 2112 kb)
10103_2014_1699_Fig7_ESM.gif (59 kb)

(GIF 58 kb)

10103_2014_1699_MOESM3_ESM.tif (896 kb)
Table 2 Histological scoring system (TIFF 896 kb)
10103_2014_1699_Fig8_ESM.gif (19 kb)

(GIF 19 kb)


  1. 1.
    Mamidi MK, Dey S, Bin Abdullah BJ, Zakaria Z, Rao MS, Das AK (2012) Cell therapy in critical limb ischemia: current developments and future progress. Cytotherapy 14:902–916PubMedCrossRefGoogle Scholar
  2. 2.
    Kyriakides TR (2009) The role of thrombospondins in wound healing, ischemia, and the foreign body reaction. J Cell Commun Signal 3:215–225PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Huang JI, Jones NF, Zhu M, Lorenz HP (2004) Chondrogenic potential of multipotential cells from human adipose tissue. Plast Reconstr Surg 113:585–594PubMedCrossRefGoogle Scholar
  4. 4.
    Gimble J (2003) Adipose-derived adult stem cells: isolation, characterization, and differentiation potential. Cytotherapy 5:362–369PubMedCrossRefGoogle Scholar
  5. 5.
    Gonzalez-Rey E, González MA, Rico L, Büscher D, Delgado M (2009) Human adult stem cells derived from adipose tissue protect against experimental colitis and sepsis. Gut 58:929–939PubMedCrossRefGoogle Scholar
  6. 6.
    Sumi M, Toya N, Yanaga K, Ohki T, Nagai R (2007) Transplantation of adipose stromal cells, but not mature adipocytes, augments ischemia-induced angiogenesis. Life Sci 80:559–565PubMedCrossRefGoogle Scholar
  7. 7.
    Harada Y, Tsujimoto S, Matsugami H, Yoshida A, Hisatome I (2013) Transplantation of freshly isolated adipose tissue-derived regenerative cells enhances angiogenesis in a murine model of hind limb ischemia. Biomed Res 34:23–29PubMedCrossRefGoogle Scholar
  8. 8.
    Heydarkhan-Hagvall S, Yang JQ, Heydarkhan S, Xu Y, Zuk PA (2008) Human adipose stem cells: a potential cell source for cardiovascular tissue engineering. Cells Tissues Organs 187:263–274PubMedCrossRefGoogle Scholar
  9. 9.
    Bhang SH, La WG, Lee TJ, Yang HS, Sun AY, Baek SH, Rhie JW, Kim BS (2011) Angiogenesis in ischemic tissue produced by spheroid grafting of human adipose-derived stromal cells. Biomaterials 32:2734–2747PubMedCrossRefGoogle Scholar
  10. 10.
    Park In S, Sang-Heon K (2013) A novel three-dimensional adipose-derived stem cell cluster for vascular regeneration in ischemic tissue. Cytotherapy 13:00681–00686Google Scholar
  11. 11.
    Hawkins D (2006) Effect of multiple exposures of low-level laser therapy on the cellular responses of wounded human skin fibroblasts. Photomed Laser Surg 24:705–714PubMedCrossRefGoogle Scholar
  12. 12.
    Hu WP, Yu CL, Lan CC, Chen GS, Yu HS (2007) Helium-neon laser irradiation stimulates cell proliferation through photostimulatory effects in mitochondria. J Invest Dermatol 127:2048–2057PubMedCrossRefGoogle Scholar
  13. 13.
    Alghamdi KM, Moussa NA (2012) Low-level laser therapy: a useful technique for enhancing the proliferation of various cultured cells. Lasers Med Sci 27:237–249PubMedCrossRefGoogle Scholar
  14. 14.
    Whelan HT, Buchman EV, Whelan NT, Turner SG, Margolis DA, Cevenini V, Stinson H, Ignatius R, Martin T, Cwiklinski J, Philippi AF, Graf WR, Hodgson B, Gould L, Kane M, Chen G, Caviness J (2001) Effect of NASA light-emitting diode irradiation on wound healing. J Clin Laser Med Surg 19(6):305–314PubMedCrossRefGoogle Scholar
  15. 15.
    Mvula B, Moore T, Abrahamse H (2008) The effect of low level laser irradiation on adult human adipose derived stem cells. Lasers Med Sci 23:277–282PubMedCrossRefGoogle Scholar
  16. 16.
    Hou JF, Yuan X, Li J, Wei YJ, Hu SS (2008) In vitro effects of low-level laser irradiation for bone marrow mesenchymal stem cells: proliferation, growth factors secretion and myogenic differentiation. Lasers Surg Med 40:726–733PubMedCrossRefGoogle Scholar
  17. 17.
    Zuk PA, Mizuno H, Huang J, Futrell JW, Katz AJ (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng Part A 7:211–228CrossRefGoogle Scholar
  18. 18.
    Tapp H, Patt JC, Gruber HE (2009) Adipose-derived stem cells: characterization and current application in orthopaedic tissue repair. Exp Biol Med 234:1–9CrossRefGoogle Scholar
  19. 19.
    Cai L, Cook TG, Liang Z, Traktuev D, Cornetta K (2007) Suppression of hepatocyte growth factor production impairs the ability of adipose-derived stem cells to promote ischemic tissue revascularization. Stem Cells 25:3234–3243PubMedCrossRefGoogle Scholar
  20. 20.
    Nie C, Morris SF (2009) Local delivery of adipose-derived stem cells via acellular dermal matrix as a scaffold: a new promising strategy to accelerate wound healing. Med Hypotheses 72:679–682PubMedCrossRefGoogle Scholar
  21. 21.
    Karsan A, Poirier GG, Zhou P, Craig R, Harlan JM (1997) Fibroblast growth factor-2 inhibits endothelial cell apoptosis by Bcl-2-dependent and independent mechanisms. Am J Pathol 151(6):1775–1784PubMedCentralPubMedGoogle Scholar
  22. 22.
    Badorff C, Popp R, Rupp S, Urbich C, Dimmeler S (2003) Transdifferentiation of blood-derived human adult endothelial progenitor cells into functionally active cardiomyocytes. Circulation 107:1024–1032PubMedCrossRefGoogle Scholar
  23. 23.
    Miyahara Y, Kataoka M, Yanagawa B, Tanaka K, Hao H (2006) Monolayered mesenchymal stem cells repair scarred myocardium after myocardial infarction. Nat Med 12:459–465PubMedCrossRefGoogle Scholar
  24. 24.
    Nakagami H, Morishita R, Iguchi S, Nishikawa T, Takami Y (2005) Novel autologous cell therapy in ischemic limb disease through growth factor secretion by cultured adipose tissue-derived stromal cells. Arterioscler Thromb Vasc Biol 25:2542–2547PubMedCrossRefGoogle Scholar
  25. 25.
    Botusan IR, Savu O, Catrina AI, Grünler J, Poellinger L, Brismar K, Catrina SB (2008) Stabilization of HIF-1a is critical to improve wound healing in diabetic mice. Proc Natl Acad Sci U S A 105:19426–19431PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    Peplow PV, Ryan B, Baxter GD (2011) Laser photobiomodulation of gene expression and release of growth factors and cytokines from cells in culture: a review of human and animal studies. Photomed Laser Surg 29:285–304PubMedCrossRefGoogle Scholar
  27. 27.
    Zaidi M, Jones DW, Pritchard KA Jr, Struve J, Nandedkar SD, Lohr NL, Pagel PS, Weihrauch D (2013) Transient repetitive exposure to low level light therapy enhances collateral blood vessel growth in the ischemic hindlimb of the tight skin mouse. Photochem Photobiol 89(3):709–713PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag London 2015

Authors and Affiliations

  • In-Su Park
    • 1
  • Arindam Mondal
    • 1
  • Phil-Sang Chung
    • 1
    • 2
  • Jin Chul Ahn
    • 1
    • 3
    • 4
  1. 1.Beckman Laser Institute KoreaDankook UniversityCheonanSouth Korea
  2. 2.Department of Otolaryngology-Head and Neck Surgery, College of MedicineDankook UniversityCheonanSouth Korea
  3. 3.Department of Biomedical ScienceDankook UniversityCheonanSouth Korea
  4. 4.Biomedical Translational Research InstituteDankook UniversityCheonanSouth Korea

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