Effect of Amniotic Membrane Extract on Cultured Human Nasal Mucosa Fibroblasts

PURPOSE: To investigate the effects of amniotic membrane extract (AME) on the survival of cultured human nasal mucosa fibroblasts. METHODS: Primary cultured human nasal mucosa fibroblasts were exposed to 0, 10, 20, or 30 microg/mL AME for 3 days. The survival of the human nasal mucosa fibroblasts was measured using the MTT assay and apoptosis was evaluated with flow cytometry using annexin-V/propidium iodide double staining. RESULTS: AME decreased significantly in fibroblast proliferation after exposure to 10 microg/mL (p = 0.000), and caused significant apoptosis of the fibroblasts after exposure to 10 microg/mL (p = 0.024). CONCLUSIONS: AME decreased fibroblast proliferation in vitro at least through induction of apoptosis. Therefore, adjuvant use of AME during endonasal dacryocystorhinostomy may improve clinical outcomes.

Effect of Mitomycin C and 5-Fluorouracil on Cultured Human Nasal Mucosa Fibroblasts

PURPOSE: To investigate the effects of two antimetabolites, mitomycin C (MMC) and 5-fluorouracil (5-FU), on proliferation of cultured human nasal mucosa fibroblasts. METHODS: Human nasal mucosa fibroblasts were primarily cultured, and exposed to various concentrations of MMC and 5-FU for 5 minutes. Control fibroblasts were exposed to only DMEM media without the drugs. Effect of drugs on cell morphology was observed by phase-contrast microscopy. Cell viability and apoptosis were measured using MTT [3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl-tetrazolium bromide] assay and Acridine orange/Hoechst (AO/HO) staining, respectively. RESULTS: In both experimental groups exposed to MMC and 5-FU, fibroblasts maintained standard spindle shape. The MTT assay showed that both MMC and 5-FU inhibited fibroblast proliferation in a dose dependent manner. AO/HO staining showed apoptotic cells in both experimental groups. CONCLUSIONS: Both MMC and 5-FU have an antiproliferative effect on fibroblasts in vitro at least through induction of apoptosis. Therefore, adjuvant use of either MMC or 5-FU during endonasal dacryocystorhinostomy may improve the clinical outcome by inhibiting proliferation of the nasal mucosa.