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THE PROMOTIVE EFFECTS
OF THERAPEUTIC ULTRASOUND DEVICE
ON WOUND HEALING
Purpose of the Study
 Evaluation of the ultrasound effects on cell physiology to
apply in wound treatment for not only debridement but also
wound healing.
CS Pak M.D., JH Jeong M.D., Ph.D., CY Heo M.D., Ph.D.
Seoul National University Bundang Hospital
Seongnam, Southkorea
Contact email: lionheo@gmail.com (CY Heo)
1
of 5
Introduction
 Ultrasound
– A mechanical vibration with a frequency greater than the upper
limit of human hearing, above approximately 20 kHz.
– Applied in many different fields including diverse clinical fields.
– An invaluable tool for diagnosis
– Could be used therapeutically in accelerating wound and bone
healing, particularly.
Methods
 Analysis of infection status:
– Bacterial number determination in wound compared with
before and after debridement by ultrasound using blood agar
culture method.
 Evaluation of cell physiological effects:
Human fibroblast and endothelial cell lines used.
– MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium
bromide) assay to measure the proliferative activities of the
cells.
– NO (nitric oxide) metabolite detection by Griess reaction.
– Collagen contents analysis through hydroxyproline presence.
– RT-PCR (reverse transcription polymerase chain reaction)
analysis to confirm gene expression level of growth factors and
cytokine
2
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Results I
3
Fig. 1 Safety of Ultrasound.
• Proliferation assay in human
fibroblasts and endothelial
cells treated with ultrasound.
• An increased activity of
proliferation in the
ultrasound-treated fibroblasts
and endothelial cells.
• Increased proliferative
activity in a time-dependent
manner.
Fig. 2 Proliferative Activity of the Cells. A: Fibroblasts; B: Endothelial Cells.
• Promotive effects of
ultrasound on raising
concentration levels of NO
(nitric oxide) metabolite, NO2
(nitrite), which plays a role in
vasodilation in the fibroblasts
and endothelial cells.
• Increasing NO production in
fibroblasts and endothelial
cells might be correlated with
collagen synthesis and neo-
or revascularization,
respectively.
Fig. 3 NO Production of the Cells. A: Fibroblasts; B: Endothelial Cells.
of 5
Results II
4
• Confirmation of bFGF (basic
fibroblast growth factor),
PDGF (platelet-derived
growth factor), and GM-CSF
(granulocyte macrophage
colony-stimulating factor)
expression in the fibroblasts.
• Increased expression levels
of bFGF, PDGF, and GM-CSF
in fibroblasts after ultrasound
treatment.
Fig. 4 Expression of the Growth Factors and Cytokine.
Fig. 5 Collagen Synthesis of
the Fibroblasts.
• Increase of collagen contents in the
ultrasound-treated fibroblasts.
• This result might be correlated with increased
NO production.
• Significant decrease of bacterial number which is colonized
in the wound area after debridement by ultrasound device.
0
Fig. 6 Debridement Effect of Ultrasound
on Bacterial Number.
of 5
Results in Summary
 Significant decrease of bacterial number which is colonized
in the wound area after debridement by ultrasound device.
 Higher proliferation rates of human fibroblasts and
endothelial cells treated with the intensity of 113W/cm2
ultrasound compared to them without ultrasound treatment.
 Promoted NO production in both cell types treated with
ultrasound.
 Increase of the expression levels of bFGF, PDGF, and GM-
CSF.
 Increase of the collagen contents in the ultrasound-treated
fibroblasts.
Conclusion
 Promoting proliferation of human fibroblasts and
endothelial cells.
 Induction of NO and collagen synthesis.
 Increasing expression of growth factors and cytokines which
are needed for wound healing process.
 These findings reveal that application of ultrasound in
wound treatment provides regenerative effects by
inducing cell proliferation and synthesis of extracellular
matrix probably including angiogenesis, and vasodilation
for promoting wound repair.
5

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EWMA 2014 - EP413 THE PROMOTIVE EFFECTS OF THERAPEUTIC ULTRASOUND DEVICE ON WOUND HEALING

  • 1. of 5 THE PROMOTIVE EFFECTS OF THERAPEUTIC ULTRASOUND DEVICE ON WOUND HEALING Purpose of the Study  Evaluation of the ultrasound effects on cell physiology to apply in wound treatment for not only debridement but also wound healing. CS Pak M.D., JH Jeong M.D., Ph.D., CY Heo M.D., Ph.D. Seoul National University Bundang Hospital Seongnam, Southkorea Contact email: lionheo@gmail.com (CY Heo) 1
  • 2. of 5 Introduction  Ultrasound – A mechanical vibration with a frequency greater than the upper limit of human hearing, above approximately 20 kHz. – Applied in many different fields including diverse clinical fields. – An invaluable tool for diagnosis – Could be used therapeutically in accelerating wound and bone healing, particularly. Methods  Analysis of infection status: – Bacterial number determination in wound compared with before and after debridement by ultrasound using blood agar culture method.  Evaluation of cell physiological effects: Human fibroblast and endothelial cell lines used. – MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay to measure the proliferative activities of the cells. – NO (nitric oxide) metabolite detection by Griess reaction. – Collagen contents analysis through hydroxyproline presence. – RT-PCR (reverse transcription polymerase chain reaction) analysis to confirm gene expression level of growth factors and cytokine 2
  • 3. of 5 Results I 3 Fig. 1 Safety of Ultrasound. • Proliferation assay in human fibroblasts and endothelial cells treated with ultrasound. • An increased activity of proliferation in the ultrasound-treated fibroblasts and endothelial cells. • Increased proliferative activity in a time-dependent manner. Fig. 2 Proliferative Activity of the Cells. A: Fibroblasts; B: Endothelial Cells. • Promotive effects of ultrasound on raising concentration levels of NO (nitric oxide) metabolite, NO2 (nitrite), which plays a role in vasodilation in the fibroblasts and endothelial cells. • Increasing NO production in fibroblasts and endothelial cells might be correlated with collagen synthesis and neo- or revascularization, respectively. Fig. 3 NO Production of the Cells. A: Fibroblasts; B: Endothelial Cells.
  • 4. of 5 Results II 4 • Confirmation of bFGF (basic fibroblast growth factor), PDGF (platelet-derived growth factor), and GM-CSF (granulocyte macrophage colony-stimulating factor) expression in the fibroblasts. • Increased expression levels of bFGF, PDGF, and GM-CSF in fibroblasts after ultrasound treatment. Fig. 4 Expression of the Growth Factors and Cytokine. Fig. 5 Collagen Synthesis of the Fibroblasts. • Increase of collagen contents in the ultrasound-treated fibroblasts. • This result might be correlated with increased NO production. • Significant decrease of bacterial number which is colonized in the wound area after debridement by ultrasound device. 0 Fig. 6 Debridement Effect of Ultrasound on Bacterial Number.
  • 5. of 5 Results in Summary  Significant decrease of bacterial number which is colonized in the wound area after debridement by ultrasound device.  Higher proliferation rates of human fibroblasts and endothelial cells treated with the intensity of 113W/cm2 ultrasound compared to them without ultrasound treatment.  Promoted NO production in both cell types treated with ultrasound.  Increase of the expression levels of bFGF, PDGF, and GM- CSF.  Increase of the collagen contents in the ultrasound-treated fibroblasts. Conclusion  Promoting proliferation of human fibroblasts and endothelial cells.  Induction of NO and collagen synthesis.  Increasing expression of growth factors and cytokines which are needed for wound healing process.  These findings reveal that application of ultrasound in wound treatment provides regenerative effects by inducing cell proliferation and synthesis of extracellular matrix probably including angiogenesis, and vasodilation for promoting wound repair. 5