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Rafael S. Arcieri, José M. Costa, Sérgio A. Costa, Danilo H. Costa, Karla O. Murta, Cecílio P. Souza Neto, Enyr S. Arcieri; Comparison of Two Ways of Preserving Sclera Samples: Dehydration in Ethyl Alcohol versus Glycerol. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4549.
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To compare the effect of preserving sclera samples in either dehydration in ethyl alcohol or glycerol.
Sixteen samples of human sclera were studied. Half of them were preserved with glycerol and half preserved by dehydration with ethyl alcohol. The weight of each sample was standardized by precision balance with 3mg. The process of protein extraction was performed using a lysis buffer, macerating and, subsequently, with beta-mercaptoethanol. After performing the whole procedure, the supernatant was removed. Eight samples were selected in which the protein was close to perform electrophoresis of proteins. Samples run on SDS-PAGE 14% gels. Subsequently, the bands were ruddy by Coomassie Blue R-250-0.5% and Silver coloration. The results were evaluated by the concentration of protein in each band of the gels in a qualitative way.
Although the protein concentration was higher in samples preserved in glycerol (31.59 ± 9.89 mg/ml) than in dehydration (27.38 ± 6.28 mg/ml), there was no statistically significant difference between groups (p=0.3262). The mean absorbance was 0.211 ± 0.066 for glycerol and 0.183 ± 0.042 for dehydration (p=0.3253). Through qualitative analysis of ruddy bands on each gel, both in Comassie-Blue and Silver staining, a higher concentration of protein was observed in the bands that ran sclera materials preserved in glycerin.
Surgeons who use sclera in ophthalmic procedures should be aware of the different characteristics when comparing the sclera preserved using glycerol or by dehydration. Maybe it could interfere with immunological response after surgery. Further studies are necessary to elucidate these findings.
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