Journal of cataract and refractive surgery · 2019
Osmotically induced removal of lens epithelial cells to prevent PCO after pediatric cataract surgery: Pilot study to assess feasibility.
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Zhang JinJun, Hussain Ali, Yue Sun, Zhang Tao, Marshall John
Аннотация
PURPOSE: Development of an osmotic-shock technique to remove human lens epithelial cells (LECs) as a preventive measure to address posterior capsule opacification (PCO) after pediatric cataract surgery.
SETTING: Department of Genetics, UCL Institute of Ophthalmology, London, England, and Department of Ophthalmology, Ruijin Hospital, Jiao Tong University, Shanghai, China.
DESIGN: Laboratory study.
METHODS: Various tissue preparations of human LECs (cultured on coverslips/collagen-coated membrane inserts, human lens capsule biopsies, and lens organ cultured PCO models) were subjected to a single or incremental hyperosmotic shock (NaCl, 350-4000 mOsm/L) in the presence of inhibitors of the Na+-K+-2Cl- cotransporter (NKCC) (to disable the regulatory volume increase [RVI] process). The integrity of the cell monolayer was determined by phase-contrast microscopy, viability assays, and measurement of transepithelial resistance.
RESULTS: Hyperosmotic shock (400 mOsm/L) caused rapid cell shrinkage (<5 minutes) in all the LEC models studied. In the absence of the NKCC inhibitor, the shrunk cells gradually returned to their original cell volume and architecture over time, while still exposed to the hyperosmotic shock. However, inhibition of the RVI process disabled the ability for restoration of cell volume leading to persistent cell shrinkage, subsequently resulting in cell detachment from the underlying support medium.
CONCLUSION: Hyperosmotic shock in the presence of inhibitors of the RVI process was effective in rapidly detaching LECs from their basement membranes. This technique could potentially facilitate removal of residual LECs left on the lens capsule after cataract surgery, thus decreasing or eliminating the risk for aggressive cell proliferation and the development of PCO.
SETTING: Department of Genetics, UCL Institute of Ophthalmology, London, England, and Department of Ophthalmology, Ruijin Hospital, Jiao Tong University, Shanghai, China.
DESIGN: Laboratory study.
METHODS: Various tissue preparations of human LECs (cultured on coverslips/collagen-coated membrane inserts, human lens capsule biopsies, and lens organ cultured PCO models) were subjected to a single or incremental hyperosmotic shock (NaCl, 350-4000 mOsm/L) in the presence of inhibitors of the Na+-K+-2Cl- cotransporter (NKCC) (to disable the regulatory volume increase [RVI] process). The integrity of the cell monolayer was determined by phase-contrast microscopy, viability assays, and measurement of transepithelial resistance.
RESULTS: Hyperosmotic shock (400 mOsm/L) caused rapid cell shrinkage (<5 minutes) in all the LEC models studied. In the absence of the NKCC inhibitor, the shrunk cells gradually returned to their original cell volume and architecture over time, while still exposed to the hyperosmotic shock. However, inhibition of the RVI process disabled the ability for restoration of cell volume leading to persistent cell shrinkage, subsequently resulting in cell detachment from the underlying support medium.
CONCLUSION: Hyperosmotic shock in the presence of inhibitors of the RVI process was effective in rapidly detaching LECs from their basement membranes. This technique could potentially facilitate removal of residual LECs left on the lens capsule after cataract surgery, thus decreasing or eliminating the risk for aggressive cell proliferation and the development of PCO.