J Korean Ophthalmol Soc.  2015 Dec;56(12):1939-1944. 10.3341/jkos.2015.56.12.1939.

Effect of Amniotic Membrane Extract on Cultured Human Nasal Mucosa Fibroblasts

• Hwang SJ
• Kim KH

• KMID: 2148758
• DOI: http://doi.org/10.3341/jkos.2015.56.12.1939

Abstract

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.

Keyword

Amniotic membrane extract; Apoptosis; Nasal mucosa fibroblast

MeSH Terms

Amnion*
Apoptosis
Dacryocystorhinostomy
Fibroblasts*
Flow Cytometry
Humans*
Nasal Mucosa*

Figure

• 

  Figure 1. Effect of amniotic membrane extract (AME) on the survival of the cultured human nasal mucosa fibroblasts. AME decreased fibroblast proliferation significantly in a dose-de-pendent manner compared to non-exposed control. * p < 0.05.

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  Figure 2. Flow cytometric analysis of apoptosis using annexin-PI double staining. Cells in quadrant B1, B2, B3, B4 represents ne-crotic cells, late apoptotic cells, living cells and early apoptotic cells, respectively. (A) Unstained control. (B) Exposed to 20 μ g/mL amniotic membrane extract. FITC = fluorescein isothiocyanate; PI = propidium iodide.

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  Figure 3. Flow cytometric analysis of apoptosis using annex-in-PI double staining. 10, 20, 30 μ g/mL amniotic membrane extract increased the degree of apoptosis significantly com-pared to non-exposed control. PI = propidium iodide.* p < 0.05.

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  Figure 4. Flow cytometric analysis of necrosis using annex-in-PI double staining. 20, 30 μ g/mL amniotic membrane ex-tract affected the degree of necrosis significantly compared to non-exposed control. PI = propidium iodide. * p < 0.05.

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