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Influence of Low Intensity Laser Irradiation on Isolated Human Adipose Derived Stem Cells Over 72 Hours and Their Differentiation Potential into Smooth Muscle Cells Using Retinoic Acid

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Abstract

Introduction

Human adipose derived stem cells (hADSCs), with their impressive differentiation potential, may be used in autologous cell therapy or grafting to replace damaged tissues. Low intensity laser irradiation (LILI) has been shown to influence the behaviour of various cells, including stem cells.

Aims

This study aimed to investigate the effect of LILI on hADSCs 24, 48 or 72 h post-irradiation and their differentiation potential into smooth muscle cells (SMCs).

Methodology

hADSCs were exposed to a 636 nm diode laser at a fluence of 5 J/cm2. hADSCs were differentiated into SMCs using retinoic acid (RA). Morphology was assessed by inverted light and differential interference contrast (DIC) microscopy. Proliferation and viability of hADSCs was assessed by optical density (OD), Trypan blue staining and adenosine triphosphate (ATP) luminescence. Expression of stem cell markers, β1-integrin and Thy-1, and SMC markers, smooth muscle alpha actin (SM-αa), desmin, smooth muscle myosin heavy chain (SM-MHC) and smoothelin, was assessed by immunofluorescent staining and real-time reverse transcriptase polymerase chain reaction (RT-PCR).

Results

Morphologically, hADSCs did not show any differences and there was an increase in viability and proliferation post-irradiation. Immunofluorescent staining showed expression of β1-integrin and Thy-1 72 h post-irradiation. RT-PCR results showed a down regulation of Thy-1 48 h post-irradiation. Differentiated SMCs were confirmed by morphology and expression of SMC markers.

Conclusion

LILI at a wavelength of 636 nm and a fluence of 5 J/cm2 does not induce differentiation of isolated hADSCs over a 72 h period, and increases cellular viability and proliferation. hADSCs can be differentiated into SMCs within 14 days using RA.

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Acknowledgements

This project was supported by the National Research Foundation (NRF) of South Africa, Council for Scientific and Industrial Research (CSIR) of South Africa, and the University of Johannesburg (UJ). Lasers were supplied and set up by the National Laser Centre (NLC) of South Africa.

Disclosures

The authors indicate no potential conflict of interest.

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Authors and Affiliations

  1. Laser Research Centre, Faculty of Health Sciences, University of Johannesburg, P.O. Box 17011, Doornfontein, 2028, South Africa

    Jennifer Anne de Villiers, Nicolette Nadene Houreld & Heidi Abrahamse

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  1. Jennifer Anne de Villiers
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  2. Nicolette Nadene Houreld
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  3. Heidi Abrahamse
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Correspondence to Heidi Abrahamse.

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de Villiers, J.A., Houreld, N.N. & Abrahamse, H. Influence of Low Intensity Laser Irradiation on Isolated Human Adipose Derived Stem Cells Over 72 Hours and Their Differentiation Potential into Smooth Muscle Cells Using Retinoic Acid. Stem Cell Rev and Rep 7, 869–882 (2011). https://doi.org/10.1007/s12015-011-9244-8

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