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

Abstract

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.

Keywords

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

Supplementary material

10103_2014_1699_MOESM1_ESM.tif (1.9 mb)
Figure 1Immunofluorescence 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 1List 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 2Histological scoring system (TIFF 896 kb)
10103_2014_1699_Fig8_ESM.gif (19 kb)

(GIF 19 kb)

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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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