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 Amniotic Membrane Extract on the Tenon's Capsule Fibroblasts

PURPOSE: To investigate the effect of amniotic membrane extract (AME) on the survival, nitric oxide production by cultured human Tenon's capsule fibroblasts (HTCF) and contraction of collagen gels. METHODS: After exposure to 5~20 g/ml AME for 5 days, the survival and nitrite production of the primarily cultured HTCF were assessed with MTT and Griess assays respectively. Acridine orange/Hoechst 33342 double staining and flow cytometric analysis using FITC-Annexin/propidium iodide double staining were done to evaluate AME-induced apoptosis of HTCF. The effect of AME on the contraction of collagen gels cultured with fibroblast was also evaluated. RESULTS: AME decreased cellular survival in a dose-dependent manner and did not affect the production of nitric oxide. AME caused apoptotic death of HTCF in a dose-dependent manner and caused necrosis at high concentration (20 g/ml). AME did not affect on the contraction of collagen gel. CONCLUSIONS: AME decreased cellular survival of HTCF by apoptosis and did not affect the contraction of collagen gels. Thus AME could modulate conjunctival wound healing by suppressing the cellular survival and activity of HTCF.

Therapeutic Effect of Amniotic Membrane Extract on Keratitis Following Corneal Alkali Burn

PURPOSE: Amniotic membrane (AM) contains various proteinase inhibitors and when used as a graft, it could enhance healing process by blocking insult of inflammatory cells and inhibiting proteolytic damage. Thus we evaluated whether applying of amniotic membrane extract as eyedrops could get the same effect as amniotic membrane patching. METHODS: Alkali wounds were inflicted on the central corneas of rabbits by applying a round filter paper, 6.0 mm in diameter, soaked in 1N NaOH for 30 seconds. A total of 16 rabbits were divided into four groups: (1) applied with amniotic membrane extract; (2) applied with amniotic membrane extract and Healon(R); (3) applied with methylcellulose; and (4) control. Each material was applied for 1 week. During follow-ups, epithelial defects, corneal thickness and its opacity were measured. RESULTS: The epithelial healing was faster and the corneal thickness was thinner in amniotic membrane extract applied groups than in non-applied. Corneal opacity was much less in AM extract applied groups. CONCLUSIONS: AM extract as eyedrops promotes wound healing and it could be an effective method for treating various keratitis due to its convenience and good effect.