April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Free-Radical Formation during Vitrectomy Vitreous Substitutes
Author Affiliations & Notes
  • Nathan Ravi
    Ophthalmolgy, Washington University School of Medicine, St Louis, MO
    Chemical Engineering, Washington University, St. Louis, MO
  • Paul David Hamilton
    Ophthalmolgy, Washington University School of Medicine, St Louis, MO
    Research, VA Health Care System, St. Louis, MO
  • Footnotes
    Commercial Relationships Nathan Ravi, None; Paul Hamilton, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 2318. doi:
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      Nathan Ravi, Paul David Hamilton, Retina; Free-Radical Formation during Vitrectomy Vitreous Substitutes. Invest. Ophthalmol. Vis. Sci. 2014;55(13):2318.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

Complications such as nuclear sclerotic cataract and glaucoma are associated with vitrectomy in elderly patients. Increased oxygen tension in the vitreous cavity is associated with these complications, but causality is unclear. We have reported that significant hydroxyl free-radicals (OHFR) were formed during cutting of biomimetic vitreous hydrogels (ARVO #2144, 2013), and may provides an alternative mechanism of tissue damage during the vitrectomy. We have supplement previous data and have studied the quenching of free-radicals by naturally occurring anti-oxidants.

 
Methods
 

Synthetic acrylamide/acrylic acid cross-linked hydrogels were used as vitreous substitutes during vitrector cutting in the presence and absence of anti-oxidants. A cut rate 3000 cuts per minute was used, with variable suction pressures. Free-radical formation was followed by electron spin resonance (ESR), using 5,5-Dimethyl-1-Pyrroline-N-Oxide (DMPO) as a spin-trap and a magnetic field strength of 330-338 milli tesla (mT), focusing on the free-radical peaks between 333-334 mT. Separately, OHFR were calibrated using 40 uM FeSO4 plus H2O2, giving 80 uM of radicals. The anti-oxidants glutathione, lipoic acid and ascorbic acid were added at 80 uM concentration in the presence of the FeSO4 and H2O2.

 
Results
 

Measurable quantities of OHFR were produced during the hydrogel cutting over a period of 20 minutes (Fig 1). Figure 2 suggests that glutathione is more effective than lipoic acid while ascorbate eliminated the OHFR peaks but produced its own ascorbyl FR peaks. The effect of glutathione and lipoic acid was additive, and the combination of all three also eliminated the OHFR and ascorbic acid FR peaks. The ESR signal produced by cutting in the presence of anti-oxidant was also quenched

 
Conclusions
 

Cutting of macromolecules by vitrector produces free radicals. Free radical production is mitigated in the presence of anti-oxidants. Ascorbic acid radicals were observed. Results indicate that it may be important to maintain a high antioxidant environment during vitrectomy, to prevent damage to the ocular tissues, especially in ageing patients where redox values are diminished. These results need to be validated in vivo.

 
 
Figure 1. OHFR signal with respect to time of cutting.
 
Figure 1. OHFR signal with respect to time of cutting.
 
 
Figure 2. Quenching of OHFR free radical signal by ESR at 333-334 mT with anti-oxidants; lipoic acid (LA), glutathione (Glu) and ascorbic acid (Asc) singly and in combination.
 
Figure 2. Quenching of OHFR free radical signal by ESR at 333-334 mT with anti-oxidants; lipoic acid (LA), glutathione (Glu) and ascorbic acid (Asc) singly and in combination.
 
Keywords: 634 oxidation/oxidative or free radical damage  
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