Dose-Dependent Cannabidiol-Induced Elevation of Intracellular Calcium and Apoptosis in Human Articular Chondrocytes

Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society
Martina WinklmayrMartin Jakab

Abstract

Cannabidiol (CBD) is the most abundant non-psychoactive compound of Cannabis sativa extracts. Cannabinoids have been shown to exhibit anti-inflammatory, analgesic, antioxidant, neuroprotective, and anti-tumorigenic effects. In the present study, we investigated the effects of CBD on human articular chondrocytes. Cell viability was determined by Resazurin assays. Apoptosis was analyzed by annexin-V/7-actinomycin D (7-AAD) staining followed by flow cytometry. Caspase 3/7 activity was measured with caspase assays. Intracellular Ca2+ ([Ca2+ ]i ) was monitored by time-lapse fluorescence imaging. The perforated whole-cell patch-clamp technique was used for measuring the cell membrane potential. Erk1/2 phosphorylation was assessed by western blot analysis. The chondrocyte cell line C28/I2 and primary chondrocytes showed a reduced viability after treatment with concentrations of CBD greater than 4 µM. This apoptotic effect was accompanied by an increase of caspase 3/7 activity and an increase in the early apoptotic cell population. CBD elevated [Ca2+ ]i , which was accompanied by depolarization of the cell membrane potential. The increase of [Ca2+ ]i was abrogated, when Ca2+ was omitted from the bath solution, indicating an influx of e...Continue Reading

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Citations

Nov 27, 2020·International Journal of Molecular Sciences·Jakub MlostKatarzyna Starowicz
Dec 8, 2020·Frontiers in Cell and Developmental Biology·Michael KittlMartin Jakab
May 27, 2021·Current Pharmaceutical Design·Wheeler TorresValmore Bermudez

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Methods Mentioned

BETA
flow cytometry
fluorescence imaging
electrophoresis
fluorescence

Software Mentioned

TILL Photonics ‐ FEI
ImageJ
Kaluza Analysis
PatchMaster
Life Acquisition and Offline Analysis
SPSS

Related Concepts

Related Feeds

Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis

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