NANOVIBRONIX ULTRASOUND TECHNOLOGY INCREASES CYTOKERATIN 14, COLLAGEN III & GLICOSEAMINOGLYCAN (GAG) EXPRESSION IN HUMAN SKIN EXPLANTS


Background

The goal – explore the potential of NanoVibronix technology in stimulating various skin precursors in human skin explants.

The tested precursors were:

  • Ki67 (cells in mitosis)
  • Basal keratin 14
  • GAGs (Glycosaminoglycan)
  • Collagen III synthesis (located in the papillary dermis)
  • Collagen IV synthesis (located in the dermal-epidermal junction)
  • PECAM-1 (blood vessels)MethodsExplants were put into six wells culture plate and treated by one of four protocols:
    • No treatment – control
    • Stimulation with intermittent period – Intermittent mode – total of 6minutes of situation including 3 cycles with stimulation for 2 minutes and intermission of 2 minutes. Treatment was repeated at days: D1, D2, D3, D6, D7, D8 and D9.Results

Cells in mitosis

(Ki67)The number of positive cells, corresponding to each sample, is presented in the following table.Basal keratin 14

The staining is quite clear, less or more regular on keratinocytes on the basal layer.

On day 10

Untreated explants:
The staining is quite clear, less or more regular on keratinocytes on the basal layer.

Treated explants
The staining is very clear, regular on keratinocytes on the basal layer and on 1 cellular layer in supra basal layer.

3. GAGs

Untreated explants on day 0:
Neutral GAGs observed close to the dermal epidermal junction (DJE) are moderate, showing a slightly regular thin pink band.
Acid GAGs are moderately present in the papillary dermis associated to thin fibres.

On day 10

Untreated explants:
Neutral GAGs observed close to the dermal epidermal junction (DJE) are moderate, showing a slightly regular thin pink band.
Acid GAGs are very moderately present in the papillary dermis associated to thin fibres.

Treated explants
Neutral GAGs observed close to the dermal epidermal junction (DJE) are clear, showing a less or more regular thin pink band.
Acid GAGs are very slightly present in the papillary dermis associated to thin fibres.

Collagen III

Untreated explants on day 0:
The staining is very slight in all the papillary dermis.

On day 10

Untreated explants:
The staining is slight in all the papillary dermis.

Treated explants:
The staining is very moderate by area in the papillary dermis.

Collagen IV

Untreated explants on day 0:

Staining is very clear, quite regular on the dermal-epidermal junction, and very moderately present in the papillary dermis. It is clear around the dermal annexes.

On day 10

Untreated explants:

Staining is clear, less or more regular on the dermal-epidermal junction, and very moderately present in the papillary dermis. It is clear around the dermal annexes.

Treated explants:
Staining is quite clear, irregular on the dermal-epidermal junction, and very moderately present in the papillary dermis. It is clear around the dermal annexes.

6. PECAM-1

Untreated explants on day 0:
The staining is clear. Blood vessels are numerous in the papillary dermis with a moderate lumen.

On day 10

Untreated explants:
The staining is clear. Blood vessels are in a moderate number in the papillary dermis with a small lumen.

Treated explants
The staining is clear. Blood vessels are in a moderate number in the papillary dermis with a small lumen.

Quantification of epidermal thickness and CK14 intensity

Treatment with intermittent mode induces a significant increase of approx. 75% and 80% in epidermal thickness and CK14 expression in comparison to the untreated explants, respectively.

Conclusions:

According to these results, Nanovibronix stimulation a clear epidermal stimulation:

  • A clear basal cytokeratin 14 over expression.
  • A slight neutral GAGs over expression.
  • A slight collagen III over expression.
  • A clear decrease of cells in mitosis.
  • No collagen IV over expression.
  • No blood vessels stimulation.

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