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Effect of mitomycin-C on contraction and migration of human nasal mucosa fibroblasts: implications in dacryocystorhinostomy
  1. Vinay Kumar1,
  2. Mohammad Javed Ali2,
  3. Charanya Ramachandran1
  1. 1Sudhakar and Sreekanth Ravi Stem Cell Biology Laboratory, L.V. Prasad Eye Institute, Hyderabad Eye Research Foundation, Hyderabad, Telangana, India
  2. 2Dacryology Service, L.V. Prasad Eye Institute, Hyderabad Eye Research Foundation, Hyderabad, Telangana, India
  1. Correspondence to Dr Charanya Ramachandran, Sudhakar and Sreekanth Ravi Stem Cell Biology Lab, Brien Holden Eye Research Institute, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Road No 2, Banjara Hills, Hyderabad, Telangana, 500034, India; charanya{at}lvpei.org

Abstract

Aim To determine the effect of mitomycin-C (MMC) on the contraction and migration of human nasal mucosal fibroblasts (HNMFs) in vitro in order to identify the least concentration of MMC required to prevent cicatrix development following dacryocystorhinostomy (DCR).

Methods Primary cultures of HNMFs were established from nasal mucosal tissues of patients undergoing DCR. Myofibroblast transformation of HNMFs was induced using transforming growth factor-β (TGF-β1) and confirmed by immunostaining for α-smooth muscle actin (α-SMA). Collagen gel contraction assay was employed to study contraction in the presence or absence of TGF-β1 (5 and 10 ng/mL) and MMC (0.2 and 0.4 mg/mL). Scratch wound assay was employed to determine the influence of MMC treatment on cell migration. Quantification of gel contraction and wound closure was done using Image J software.

Results α-SMA expression increased with TGF-β1 treatment in a time- and dose-dependent manner indicating myofibroblast transformation of HNMFs. MMC inhibited TGF-β1- induced collagen gel contraction in a dose-dependent manner (0.4 mg/mL>0.2 mg/mL). Further, there was a decrease in the migration of MMC-treated HNMFs, resulting in delayed wound closure that corroborated with the loss of actin stress fibres.

Conclusions MMC successfully inhibited TGF-β1-induced myofibroblast transformation, collagen gel contraction and significantly reduced the migration of HNMFs to cover the wound even at a low concentration of 0.2 mg/mL. This study provides evidence that low concentration and short duration of MMC treatment is efficient in reducing increased contraction and migration of HMNFs in response to injury.

  • Lacrimal drainage
  • Wound healing

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