1. Differences in the ablation of the Great Saphenous Vein by 810nm and 1470nm Endovenous Lasers, as shown by histology and immunohistochemistry using a novel method of in-vitro treatment
Henry F. Ashpitel1, F. Javier Salguero1, Waldo García-Jiménez1,
Roberto M. La Ragione1 and Mark S. Whiteley1,2
1Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, GU2 7XH.
2The Whiteley Clinic, 1 Stirling House, Stirling Road, Guildford, Surrey, GU2 7RF.

2. Disclosures & Ethics
Henry Ashpitel is funded for his PhD by an educational grant from AngioDynamics. This work is a part of his PhD studies.
This study received favourable ethical approval from:
NHS HRA, NRES Committee South East Coast-Surrey: 13/LO/0058
University of Surrey Ethics Committee: EC/2013/23/FHMS

3. Introduction NICE CG168 (July 2013):
Treatment of varicose veins preferentially:
“Endothermal ablation”

4. Introduction EVLA Initially chromophore used haemoglobin
810 nm, 940 nm, 941 nm, 1064 nm
More recently chromophore used water
1470 nm, 1920 nm
Both 810 nm and 1470 nm commonly used
Both reported to be able to ablate GSV at
40 – 100 J/cm

5. Aims
Much clinical research into recurrence rates and patient satisfaction
Little research into biological effects of EVLA on GSV
To investigate:
The difference in the biological effect of 810 nm and 1470 nm EVL in GSV
Examine the expression of various apoptotic-linked proteins in GSV treated with the EVL’s
Develop an original model of treating ex-vivo GSV

6. Method Ten lengths of ex-vivo extra-fascial GSV:
Post-collection, tissue cultured overnight at 37˚C
Five treated with 810 nm EVL and five treated with 1470 nm EVL (AngioDynamics)
Each length treated in five sections: 0 (CTL), 20, 40, 60 and 80 J/cm
Post-treatment tissue cultured at 37˚C.
Sub-sections from each treatment group retrieved at 6 and 24 hours
Sub-sections taken for histological and immunohistochemical analysis
Histological Analysis: Haematoxylin and Eosin (H&E) and Martius Scarlet Blue (MSB)
Immunohistochemical Analysis: anti-smooth muscle actin (αSMA) and anti-caspase-3 (αC3)

7. Results - Histology MSB Staining Blue – Collagen Red – Fibrin
810 nm 40 J/cm
1470 nm 40 J/cm
Control
810 nm 60 J/cm
1470 nm 60 J/cm

8. Results - Immunohistochemistry
αSMA Staining
Brown/Red = Smooth Muscle Actin
810 nm 40 J/cm
1470 nm 40 J/cm
810 nm 60 J/cm
1470 nm 60 J/cm
Control

9. Results - Immunohistochemistry
αC3 Staining
Brown/Red = Cleaved Caspase-3
Arrows indicate Caspase-3 expression
810 nm 40 J/cm
1470 nm 40 J/cm
810 nm 60 J/cm
1470 nm 60 J/cm
Control

10. Analysis of Results
Clear that there is a significant difference between treatment with 810 nm and 1470 nm EVL’s
Significant increase of thermal damage and spread with the 1470 nm EVL at 40, 60 and 80 J/cm
Confirmed with αSMA immunohistochemical staining
More caspase-3 expression seen with the 810 nm EVL
Very little caspase-3 expression seen with the 1470 nm EVL at 6 hours and nearly none at 24 hours
Significantly less caspase-3 expression in sections treated with the 1470 nm EVL at 40, 60 and 80 J/cm compared to the control

11. Conclusions
Thermal damage and spread increased when using the 1470 nm EVL
Caspase-3 immunohistochemistry suggests:
That most of the cells in tissue treated with 1470nm EVL have undergone apoptosis by 24 hours
That most of the cells in tissue treated with 810 nm EVL are undergoing apoptosis by 24 hours
Hence, there is a clear superiority in damage and cellular apoptosis in GSV treated with 1470 nm EVL
Novel model appears to be successful and easy to use
Feedback confirmed methodology simplicity and practicality