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    Biomechanics

    Therapeutic Ultrasound Increases Vascular Endothelial Growth Factor Secretion in Endothelial Cells

    (D-153) Therapeutic Ultrasound Increases Vascular Endothelial Growth Factor Secretion in Endothelial Cells

    Friday, October 13, 2023
    3:30 PM – 4:30 PM PDT
    Location: Exhibit Hall - Row D - Poster # 153

      Presenting Author(s)

    • Enzo Ottaviani

      Undergraduate Researcher
      Bucknell University
      Wysox, Pennsylvania, United States

      • Primary Investigator(s)

    • OB

      Olivia Boerman

      Assistant Professor
      Bucknell University
      Lewisburg, Pennsylvania, United States

    Introduction:: Low Intensity Pulsed Ultrasound (LIPUS) has been shown to accelerate wound healing in chronic wounds. Angiogenesis, the

    creation of new blood vessels, is a critical step in the wound healing process. One of the key secreted proteins found to

    correlate with angiogenesis is Vascular Endothelial Growth Factor (VEGF). VEGF supports the growth and proliferation of

    endothelial cells leading to increased angiogenesis. This study was performed to understand how therapeutic ultrasound

    affects VEGF secretion in endothelial cells and ultimately angiogenesis.

    Materials and Methods::

    80,000 Human Umbilical Vein Cord (HUVEC) cells were seeded per well in a 12-well plate in 2 milliliters of media (n=3). After reaching confluency, ultrasound was applied to each well for 15 minutes at 36 kHz and intensities of 50 mW/cm2, 100 mW/cm2, and 150 mW/cm2. Conditioned media was then collected and stored at -20℃, until analysis. An Enzyme Linked Immunosorbent Assay (ELISA) was used to quantify VEGF secretion. ELISAs are performed by using a capture antibody, detection antibody, and substrate-each used to make the VEGF attach to the plate and be shown using a plate reader at 450 nanometer and 605 nanometers.


    After the data was collected, the absorbance values collected at 605 nanometers were subtracted from the absorbance values at 450 nanometers. Duplicates were then averaged together, and the standard curve was fitted with an exponential line of best fit generating an equation. This equation was then used to calculate the unknown concentrations of samples in pg/mL. A single factor ANOVA was performed with Tukey Post Hoc analysis. A p-value less than 0.05 was considered significant.



    Results, Conclusions, and Discussions::

    We found a significant difference in VEGF secretion between the control group and ultrasound intensity of 150 mW/cm2. It was also shown that there was a significant amount of more VEGF secreted in the 100 mW/cm2 than the 50 mW/cm2, as well as the 150 mW/cm2 compared to the 50 mW/cm2 group.


    The 150 mW/cm2 was found to be significantly higher than the control. This suggests that 150mW/cm2 intensity of therapeutic ultrasound may increase VEGF secretion leading to increased angiogenesis. As the intensity increased, there were significantly higher concentrations of VEGF at 100 mW/cm2 and 150 mW/cm2 compared to the intensity of 50 mW/cm2. The most significant difference between all of the groups was between the 50 mW/cm2 and 150 mW/cm2 ultrasound intensity groups.


    This data suggests that in the context of VEGF secretion, a higher intensity of ultrasound would be more beneficial than a lower intensity. 50 mW/cm2 produced 500 pg/mL of VEGF secretion, while at 100 mW/cm2 average 1250 pg/mL, and at 150 mW/cm2 at 1750 pg/mL. From these amounts, it would be most beneficial to use 150 mW/cm2 for the potential of increased angiogenesis in HUVEC cells. Increased angiogenesis would lead to advanced healing of chronic wounds.


     



    Acknowledgements (Optional): :

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