Investigation of the Mitigation of DMSO-Induced Cytotoxicity by Hyaluronic Acid following Cryopreservation of Human Nucleus Pulposus Cells

To develop an off-the-shelf therapeutic product for intervertebral disc (IVD) repair using nucleus pulposus cells (NPCs), it is beneficial to mitigate dimethyl sulfoxide (DMSO)-induced cytotoxicity caused by intracellular reactive oxygen species (ROS). Hyaluronic acid (HA) has been shown to protect...

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Published in:International journal of molecular sciences 2023-08, Vol.24 (15), p.12289
Main Authors: Munesada, Daiki, Sakai, Daisuke, Nakamura, Yoshihiko, Schol, Jordy, Matsushita, Erika, Tamagawa, Shota, Sako, Kosuke, Ogasawara, Shota, Sato, Masato, Watanabe, Masahiko
Format: Article
Language:English
Subjects:
Biomechanics
Cell growth
Clinical trials
Cryopreservation
Cytotoxicity
dimethyl sulfoxide
Ethylenediaminetetraacetic acid
Health aspects
Hyaluronic acid
intervertebral disc
Mitochondrial DNA
nucleus pulposus
nucleus pulposus progenitor cell
Oxidative stress
Quality of life
Reactive oxygen species
Tie2 receptor
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Summary:To develop an off-the-shelf therapeutic product for intervertebral disc (IVD) repair using nucleus pulposus cells (NPCs), it is beneficial to mitigate dimethyl sulfoxide (DMSO)-induced cytotoxicity caused by intracellular reactive oxygen species (ROS). Hyaluronic acid (HA) has been shown to protect chondrocytes against ROS. Therefore, we examined the potential of HA on mitigating DMSO-induced cytotoxicity for the enhancement of NPC therapy. Human NPC cryopreserved in DMSO solutions were thawed, mixed with equal amounts of EDTA-PBS (Group E) or HA (Group H), and incubated for 3-5 h. After incubation, DMSO was removed, and the cells were cultured for 5 days. Thereafter, we examined cell viability, cell proliferation rates, Tie2 positivity (a marker of NP progenitor cells), and the estimated numbers of Tie2 positive cells. Fluorescence intensity of DHE and MitoSOX staining, as indicators for oxidative stress, were evaluated by flow cytometry. Group H showed higher rates of cell proliferation and Tie2 expressing cells with a trend toward suppression of oxidative stress compared to Group E. Thus, HA treatment appears to suppress ROS induced by DMSO. These results highlight the ability of HA to maintain NPC functionalities, suggesting that mixing HA at the time of transplantation may be useful in the development of off-the-shelf NPC products.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms241512289