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Li W.-H.,Affiliated Hospital of Logistic College | Gao Y.-W.,Xingtai Boai Hospital | Li D.-S.,Affiliated Hospital of Logistic College | Xing Y.-X.,Affiliated Hospital of Logistic College
Chinese Journal of Tissue Engineering Research | Year: 2013

Background: Botulinum toxin type A can be used to treat human hypertrophic scar clinically and inhibit the proliferation of human hypertrophic scar fibroblasts in vitro. Objective: To investigate the effect of botulinum toxin type A on the proliferation and apoptosis of human hypertrophic scar fibroblasts. Methods: Human hypertrophic scar fibroblasts were isolated and cultured using digestion method. The cells were treated with botulinum type A at different concentrations. After 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide dyeing, the cell solutions were detected with enzyme-linked immunosorbent detector at 570 nm. The inhibitory rate was calculated. The cultured cells were stained with hoechst 33342 and PI. The apoptotic cells were observed under fluorescence microscopy and the apoptotic rate was calculated. Results and Conclusion: The hypertrophic scar fibroblasts represented the spindle-like character with uniform composition and strong proliferation. After treated with botulinum toxin type A, the cells grew slowly, the cell amount reduced and the arrangement of the cells was scattered. The absorbance after 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide dyeing decreased. There was a statistically significant difference compared with the control group (P < 0.05). The half inhibitory rate was 0.4 IU/L. After hoechst 33342 and PI dyeing, the nuclei became blue and showed a smooth round or oval appearance. After treated with botulinum toxin type A, the nuclei were condensed with uneven dyeing. Some of the nuclei were fragmented and apoptotic bodies appeared. The apoptotic rate was increased with increasing concentrations of botulinum toxin type A. There was a statistically significant difference between the control group and the treated group (P < 0.05). The half apoptotic rate was 0.4 IU/L. These findings indicate that botulinum toxin type A can cause apoptosis of the human hypertrophic scar fibroblasts, thus inhibiting the proliferation of the cells. Source

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