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Characterisation of human orbital fibroblasts cultivated from intraconal, nasal and central adipose tissues

Abstract

Purpose To investigate the characteristics of human orbital fibroblasts (OFs) cultivated from intraconal, nasal and central adipose tissues.

Methods Intraconal adipose tissues were obtained during orbital decompression surgery for severe proptosis in nine patients with Graves’ orbitopathy (GO). Nasal and central adipose tissues were obtained during upper eyelid blepharoplasty in nine patients with no history of GO. Human OFs were separately cultured from GO intraconal, non-GO nasal, non-GO central orbital adipose deposits. Human dermal fibroblasts were also cultured from redundant resected skin tissue obtained during upper eyelid blepharoplasty in normal controls. Expression of insulin-like growth factor 1 (IGF-1) and thyroid-stimulating hormone (TSH) receptors were investigated using real-time quantitative reverse transcription PCR. Protein levels of interleukin-1β (IL-1β)-induced inflammatory cytokines and generated intracellular reactive oxygen species (ROS) were determined.

Results IGF-1 and TSH receptor RNA expressions of GO intraconal OFs and non-GO nasal OFs were higher than non-GO central OFs and dermal fibroblasts. The expression of IL-1β induced the IL-6, IL-8, intercellular adhesion molecule-1 and cyclooxygenase-2 of GO intraconal OFs, and non-GO nasal OFs were higher than non-GO central OFs and dermal fibroblasts. Intracellular ROS generation in GO intraconal OFs and non-GO nasal OFs were higher than in non-GO central OFs and dermal fibroblasts, although the differences were not statistically significant.

Conclusions Non-GO nasal OFs had similar characteristics to GO intraconal OFs. We recommend the use of nasal adipose tissue in order to culture OFs as a normal control involving in vitro experiments.

  • Graves’ orbitopathy
  • orbital adipose tissue
  • orbital fibroblast

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