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Updated: Aug 21, 2026
A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Published on: April 11, 2014
Ascorbic acid reversibly inhibits proliferation of retinal pigment epithelial cells
A Heckelen1, M Hermel, B Kondring
1Interdisciplinary Centre of Clinical Research (IZKF) BIOMAT, Technical University of Aachen, Germany.
Purpose:
Proliferation control in adult retinal pigment epithelial (ARPE) cells is an essential factor in the clinical management of proliferative vitreoretinopathy (PVR). Factors which inhibit PVR and which are without toxic potential are therefore of interest in controlling proliferation. The aim of the present study was to gain insight into a possible function of high intraocular ascorbic acid levels as a physiological modulator of proliferation.
Methods:
Adult retinal pigment epithelial cells were incubated in vitro with increasing concentrations of ascorbic acid (0.5-4 mmol, pH 7.4). Cell proliferation was assayed by the bromide-deoxy-uridine (BrdU) assay. The culture medium (CM) containing ascorbic acid was replaced with normal CM and the recovery of proliferation was measured after 24 hours. In order to be able to distinguish between proliferation inhibition, apoptosis, necrosis and recovery of proliferation, we performed TUNEL assays and fluorescence analysis cell-counter (FAC) analysis.
Results:
Ascorbic acid significantly inhibits ARPE cell proliferation if it is present in concentrations above 2 mmol. Proliferation resumed in all ARPE cell cultures after pre-incubation with ascorbic acid, indicating that direct toxicity of ascorbic acid is a negligible factor. The time-point and extent of recovery in proliferation was dependent on the initial ascorbic acid concentration. Fluorescence-labelled cell counts on apoptosis markers (FAC) data showed some induction of apoptosis and necrosis after incubation with 4 mmol ascorbic acid.
Conclusions:
Ascorbic acid has a dose-dependent influence on the proliferation of vital ARPE cells. This possibly reflects the role of ascorbic acid at a physiological level within the vitreous cavity in preventing proliferative vitreoretinopathy (PVR). These findings may stimulate the development of new strategies in the clinical treatment of PVR.
Insights
High levels of ascorbic acid inhibit adult retinal pigment epithelial cell proliferation, suggesting a role in preventing proliferative vitreoretinopathy (PVR). Recovery of proliferation occurred after ascorbic acid removal, indicating low toxicity.
Area of Science:
- Ophthalmology
- Cell Biology
- Biochemistry
Background:
- Proliferative vitreoretinopathy (PVR) is a sight-threatening condition driven by retinal pigment epithelial (ARPE) cell proliferation.
- Controlling ARPE cell proliferation is crucial for PVR management.
- Identifying non-toxic proliferation inhibitors is a key therapeutic goal.
Purpose of the Study:
- To investigate the role of intraocular ascorbic acid as a physiological modulator of ARPE cell proliferation.
- To determine if high ascorbic acid levels inhibit ARPE cell proliferation.
- To assess the potential toxicity and recovery of proliferation after ascorbic acid exposure.
Main Methods:
- Adult retinal pigment epithelial cells were cultured in vitro with varying ascorbic acid concentrations (0.5-4 mmol).
- Cell proliferation was measured using the bromide-deoxy-uridine (BrdU) assay.
- Apoptosis and necrosis were assessed via TUNEL assays and fluorescence-activated cell sorting (FACs) analysis.
Main Results:
- Ascorbic acid significantly inhibited ARPE cell proliferation at concentrations above 2 mmol.
- Proliferation resumed after ascorbic acid removal, indicating minimal direct toxicity.
- Higher ascorbic acid concentrations (4 mmol) induced some apoptosis and necrosis.
Conclusions:
- Ascorbic acid exhibits a dose-dependent inhibitory effect on ARPE cell proliferation.
- These findings support a physiological role for ascorbic acid in preventing PVR within the vitreous cavity.
- The results may inform novel therapeutic strategies for PVR treatment.
