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脉冲电刺激与电化学条件培养基对人成骨细胞作用的差异

Discrimination between the effects of pulsed electrical stimulation and electrochemically conditioned medium on human osteoblasts.

作者信息

Bielfeldt Meike, Budde-Sagert Kai, Weis Nikolai, Buenning Maren, Staehlke Susanne, Zimmermann Julius, Arbeiter Nils, Mobini Sahba, González María Ujué, Rebl Henrike, Uhrmacher Adelinde, van Rienen Ursula, Nebe Barbara

机构信息

Institute for Cell Biology, Rostock University Medical Center, 18057, Rostock, Germany.

Institute of Communications Engineering, University of Rostock, 18051, Rostock, Germany.

出版信息

J Biol Eng. 2023 Nov 23;17(1):71. doi: 10.1186/s13036-023-00393-1.


DOI:10.1186/s13036-023-00393-1
PMID:37996914
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10668359/
Abstract

BACKGROUND: Electrical stimulation is used for enhanced bone fracture healing. Electrochemical processes occur during the electrical stimulation at the electrodes and influence cellular reactions. Our approach aimed to distinguish between electrochemical and electric field effects on osteoblast-like MG-63 cells. We applied 20 Hz biphasic pulses via platinum electrodes for 2 h. The electrical stimulation of the cell culture medium and subsequent application to cells was compared to directly stimulated cells. The electric field distribution was predicted using a digital twin. RESULTS: Cyclic voltammetry and electrochemical impedance spectroscopy revealed partial electrolysis at the electrodes, which was confirmed by increased concentrations of hydrogen peroxide in the medium. While both direct stimulation and AC-conditioned medium decreased cell adhesion and spreading, only the direct stimulation enhanced the intracellular calcium ions and reactive oxygen species. CONCLUSION: The electrochemical by-product hydrogen peroxide is not the main contributor to the cellular effects of electrical stimulation. However, undesired effects like decreased adhesion are mediated through electrochemical products in stimulated medium. Detailed characterisation and monitoring of the stimulation set up and electrochemical reactions are necessary to find safe electrical stimulation protocols.

摘要

背景:电刺激用于促进骨折愈合。在电极处进行电刺激时会发生电化学过程,并影响细胞反应。我们的方法旨在区分电化学和电场对成骨样MG-63细胞的影响。我们通过铂电极施加20 Hz的双相脉冲,持续2小时。将细胞培养基的电刺激及随后施加于细胞的情况与直接刺激的细胞进行比较。使用数字孪生预测电场分布。 结果:循环伏安法和电化学阻抗谱显示电极处发生了部分电解,培养基中过氧化氢浓度增加证实了这一点。虽然直接刺激和交流预处理培养基都降低了细胞黏附和铺展,但只有直接刺激增强了细胞内钙离子和活性氧。 结论:电化学副产物过氧化氢不是电刺激细胞效应的主要贡献者。然而,诸如黏附降低等不良效应是通过受刺激培养基中的电化学产物介导的。为了找到安全的电刺激方案,有必要对刺激设置和电化学反应进行详细表征和监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/60f7bb008f15/13036_2023_393_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/5335724ebcfa/13036_2023_393_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/3f49771563a1/13036_2023_393_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/8860141f0a1e/13036_2023_393_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/f692f9e3ec4b/13036_2023_393_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/ea7f07587c6d/13036_2023_393_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/94620e4cd131/13036_2023_393_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/2756a32d2a30/13036_2023_393_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/60f7bb008f15/13036_2023_393_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/5335724ebcfa/13036_2023_393_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/3f49771563a1/13036_2023_393_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/8860141f0a1e/13036_2023_393_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/f692f9e3ec4b/13036_2023_393_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/ea7f07587c6d/13036_2023_393_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/94620e4cd131/13036_2023_393_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/2756a32d2a30/13036_2023_393_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f4b/10668359/60f7bb008f15/13036_2023_393_Fig8_HTML.jpg

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[4]
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[6]
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[7]
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本文引用的文献

[1]
Advanced electrochemical potential monitoring for improved understanding of electrical neurostimulation protocols.

J Neural Eng. 2023-6-22

[2]
Alternating electric field stimulation: Phenotype analysis and osteoclast activity of differentiated RAW 264.7 macrophages on hydroxyapatite-coated Ti6Al4V surfaces and their crosstalk with MC3T3-E1 pre-osteoblasts.

Biomater Adv. 2023-3

[3]
Long-term stimulation with alternating electric fields modulates the differentiation and mineralization of human pre-osteoblasts.

Front Physiol. 2022-9-30

[4]
DC electrical stimulation enhances proliferation and differentiation on N2a and MC3T3 cell lines.

J Biol Eng. 2022-10-13

[5]
Pulsed Electrical Stimulation Affects Osteoblast Adhesion and Calcium Ion Signaling.

Cells. 2022-8-25

[6]
Bio-piezoelectricity: fundamentals and applications in tissue engineering and regenerative medicine.

Biophys Rev. 2022-6-28

[7]
Electrical stimulation to promote osseointegration of bone anchoring implants: a topical review.

J Neuroeng Rehabil. 2022-3-21

[8]
Electronic Bone Growth Stimulators for Augmentation of Osteogenesis in and Models: A Narrative Review of Electrical Stimulation Mechanisms and Device Specifications.

Front Bioeng Biotechnol. 2022-2-14

[9]
The role of the double layer for the pseudocapacitance of the hydrogen adsorption on platinum.

Sci Rep. 2022-3-1

[10]
Using a Digital Twin of an Electrical Stimulation Device to Monitor and Control the Electrical Stimulation of Cells .

Front Bioeng Biotechnol. 2021-12-8

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