The cytotoxic effect of Rhizoma coptidis, Radix scutellariae, and Cortex phellodendri chinensis on human corneal epithelial cells (CRL-11135)

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The cytotoxic effect of Rhizoma coptidis, Radix scutellariae, and Cortex phellodendri chinensis on human corneal epithelial cells (CRL-11135)

 

Author: Yau, Kwok-cheung Philip
Title: The cytotoxic effect of Rhizoma coptidis, Radix scutellariae, and Cortex phellodendri chinensis on human corneal epithelial cells (CRL-11135)
Degree: M.Sc.
Year: 2011
Subject: Hong Kong Polytechnic University -- Dissertations
Herbs -- Therapeutic use.
Materia medica.
Department: Dept. of Health Technology and Informatics
Pages: i, 80 leaves : ill. (some col.) ; 30 cm.
InnoPac Record: http://library.polyu.edu.hk/record=b2449782
URI: http://theses.lib.polyu.edu.hk/handle/200/6142
Abstract: Previous study had shown the selected herbs, Rhizoma coptidis, Radix scutellariae and Cortex phellodendri chinensis, were potent antibiotics which could be used as preservative in ophthalmic product (Boost, 2011; O'Donoghue et al., 2008). This experiment investigated if the selected herbs were safe for human use by studying the cytotoxicity of these selected herbs on corneal epithelial cells (CRL-11135) in vitro. The above herbs were extracted with 80 ℃ hot water and then diluted with phosphate-buffered saline to 5%, 3% and 1% weight by volume (g/ml). The corneal epithelial cells were treated with the diluted extracts at 5%, 3% and 1% concentrations for 5 minutes, incubated at 37 ℃ with 5% CO₂ air. After incubation, the cell's viability was tested by Vi-cell with trypan blue staining, by spectrometer with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) metabolism testing and by flow cytometry with Annexin V-FITC and 7-AAD staining. The results were statistically analyzed by non-parametric Kruskal-Wallis test (p>0.05). If significant difference was found, then Bonferroni's Multiple Comparison Test, p<0.05, was used to determine which group was different. Testing by Vi-cell with trypan blue and spectrometer with MTT could not detect any evidence of cytotoxicity in the treatment groups of all three herbs. In flow cytometry testing, Rhizoma coptidis at 5%, 3% and 1% concentrations caused a significant number of cells to be positive in both 7-AAD and Annexin V, which signified the cells were in late stage of cell death. The mean percentages of cells treated with Rhizoma coptidis that were positive with 7-AAD and Annexin V at 5%, 3% and 1% were 79.13±7.899 %, 86.28±9.901%, and 72.78±12.64% (±SD) respectively. There was no evidence of cytotoxicity from Radix scutellariae and Cortex phellodendri chinensis by flow cytometery testing.

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