Prostaglandin F2α Agonists Negatively Modulate the Size of 3D Organoids from Primary Human Orbital Fibroblasts

Invest Ophthalmol Vis Sci. 2020 Jun 3;61(6):13. doi: 10.1167/iovs.61.6.13.

Abstract

Purpose: To elucidate the molecular etiology of deepening of the upper eyelid sulcus (DUES) induced by prostaglandin (PG) analogs, a three-dimensional (3D) tissue culture system was employed using human orbital fibroblasts (HOFs).

Methods: During adipogenesis, changes in HOF 3D organoid sizes, as well as their lipids stained by BODIPY and expression of the extracellular matrix (ECM) by immunolabeling and/or quantitative PCR, were studied in the presence or absence of either 100-nM bimatoprost acid or 100-nM prostaglandin F2α.

Results: The size of the 3D organoids increased remarkably during adipogenesis, but such increases were significantly inhibited by the presence of PG analogs. Staining intensities by BODIPY and mRNA expression of peroxisome proliferator-activated receptor gamma were significantly increased upon adipogenesis but were not influenced by the presence of PG analogs. Unique changes in ECM expression observed with or without adipogenic differentiation were significantly modified by the presence of PG analogs.

Conclusions: Our present study indicates that PG analogs have the potential to modulate the ECM network within HOF 3D organoids. Thus, a 3D tissue culture system may be a suitable strategy for understanding the disease etiology of DUES.

MeSH terms

  • Adipogenesis / physiology*
  • Antihypertensive Agents / pharmacology
  • Bimatoprost / pharmacology
  • Boron Compounds / metabolism
  • Cell Culture Techniques
  • Cells, Cultured
  • Dinoprost / agonists
  • Dinoprost / pharmacology*
  • Extracellular Matrix / metabolism
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Fluorescent Dyes / metabolism
  • Humans
  • Microscopy, Confocal
  • Orbit / cytology*
  • Organoids / metabolism*
  • PPAR gamma / genetics
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction

Substances

  • 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene
  • Antihypertensive Agents
  • Boron Compounds
  • Fluorescent Dyes
  • PPAR gamma
  • RNA, Messenger
  • Dinoprost
  • Bimatoprost