Effects of Amniotic Membrane Extract on the Hyperplastic Response of the Middle Ear Mucosa in a Bacterially-Induced Otitis Media Rat Model: A Preliminary Study
Joo Hyun Park1,2, Hee-Bok Kim1, Seo Hyun Ko1
1Department of Otorhinolaryngology-Head and Neck Surgery, Dongguk University Ilsan Hospital, Dongguk University College of Medicine, Goyang, Korea.
Objectives:
Human amniotic membrane extract (AME) is known to contain numerous bioactive factors and anti-inflammatory substances. However, the anti-inflammatory effects of AME on the middle ear (ME) mucosa are unclear. This study assessed the effects of AME on the growth of the ME mucosa in response to bacterially-induced otitis media (OM).
Methods:
OM was induced by inoculating nontypeable Haemophilus influenzae (NTHi) into the ME cavity of rats. ME mucosal explants were cultured in AME concentrations of 0, 5, 10, or 50 μg/mL. The area of explant outgrowth was measured in culture and analyzed at 1, 3, 5, and 7 days after explantation. The expression of Ki-67, mucin 5AC (MUC5AC), tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10) in the explants was also evaluated using quantitative polymerase chain reaction (PCR) and immunocytochemistry (ICC).
Results:
The NTHi-induced ME mucosa growth increased gradually over the 7-day culture period in all explants at different AME concentrations. There was a trend for mucosal growth inhibition at higher concentrations of AME, although the growth was not significantly different among the groups until day 5. The ME mucosal explants treated with the 50 μg/mL concentration of AME showed significantly suppressed growth on postexplantation day 7 compared with other explants on the same day. PCR and ICC staining revealed that the expression of Ki-67, MUC5AC, TNF-α, and IL-10 further decreased in the explants with higher concentrations of AME than in those with lower concentrations of AME.
Conclusion:
Our results showed that higher concentrations of AME reduced the mucosal proliferative response in bacterial OM in rats. These findings provide evidence that AME has an influence on the inflammatory and proliferative responses to NTHi infection in ME mucosa.
Insights
Human amniotic membrane extract (AME) can reduce middle ear mucosal growth during bacterial otitis media (OM) in rats. Higher AME concentrations suppressed inflammation and proliferation, suggesting therapeutic potential for OM treatment.
Area of Science:
- Otolaryngology
- Regenerative Medicine
- Immunology
Background:
- Human amniotic membrane extract (AME) contains bioactive factors with known anti-inflammatory properties.
- The specific effects of AME on middle ear (ME) mucosa, particularly during otitis media (OM), remain largely uncharacterized.
Purpose of the Study:
- To investigate the impact of AME on the proliferative response of ME mucosa in a rat model of bacterial OM.
- To evaluate the anti-inflammatory and proliferative effects of varying AME concentrations on ME mucosal explants.
Main Methods:
- Otitis media (OM) was induced in rats using nontypeable Haemophilus influenzae (NTHi).
- ME mucosal explants were cultured with different concentrations of AME (0, 5, 10, 50 μg/mL).
- Explant outgrowth, Ki-67, MUC5AC, TNF-α, and IL-10 expression were analyzed via quantitative PCR and immunocytochemistry.
Main Results:
- AME treatment showed a trend towards inhibiting ME mucosal growth, with significant suppression observed at 50 μg/mL by day 7.
- Higher AME concentrations led to decreased expression of proliferation marker Ki-67 and inflammatory markers TNF-α.
- Expression of MUC5AC and anti-inflammatory cytokine IL-10 also decreased with increasing AME concentrations.
Conclusions:
- Elevated concentrations of AME effectively reduced the mucosal proliferative response in a rat model of bacterial OM.
- These findings indicate that AME modulates inflammatory and proliferative processes in ME mucosa during NTHi infection.
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