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Evaluation of epinephrine and vasopressin toxicity in human mesenchymal stem cells (hMSCs). in vitro study
Evaluation of epinephrine and vasopressin toxicity in human mesenchymal stem cells (hMSCs). in vitro study
Purpose: Epinephrine is an effective vasoconstrictor, and it is commonly administered intra-articular during arthroscopy to control the bleeding as a local hemostatic agent. However, no study so far has investigated possible toxicities of either epinephrine or an alternative like vasopressin on human mesenchymal stem cells. Methods: Triple-layer culture flasks have been used to seed the hMSCs; we have seeded the cells at a density of 10,000 cells/cm2. The hMSCs were plated into 96-well plates for evaluation of mitochondrial activity or 6-well plates for evaluation of apoptosis, and all the plates contained culture medium. Cells were treated with either epinephrine or vasopressin, and the control group was untreated. Epinephrine and vasopressin were removed after forty minutes, and freshly prepared growth medium has been provided to the cells. We evaluated two apoptosis markers (CASP3 and PARP-1) and the activity of the mitochondria at one hour, twenty-four hours, and seven days. Results: Mitochondrial activity did not significantly decrease after exposure to either epinephrine or vasopressin. In the cells exposed to epinephrine, we noted a significant increase in PARP-1 level at seven days; we noted the same increase after exposure to vasopressin at twenty-four hours and seven days. The amount of CASP3 has significantly increased only at twenty-four hours in those cells which were exposed to epinephrine. Conclusions: These data suggest that neither epinephrine nor vasopressin influence mitochondrial activity in hMSCs in vitro. We observed an increase in apoptosis markers without a corresponding decrease in mitochondrial activity, which could indicate that both medications are toxic to hMSCs. The absence of a response of CASP3 after exposure to the vasopressin, suggest that vasopressin may be less toxic for hMSCs. However, while epinephrine and vasopressin are effective as hemostatic agents, they may have toxic effects on hMSCs, warranting caution in their use during arthroscopy, especially if performing the kind of procedure to induce the tissue repair and regeneration (for example, meniscal-repair, microfracture, tendon-repair, OATS, ACI, etc.).
cell toxicity, human mesenchymal stem cells, local hemostatic agents, intra-articular hemostatis, intra-articular bleeding control
Akasheh, Amirpoya
2021
English
Universitätsbibliothek der Ludwig-Maximilians-Universität München
Akasheh, Amirpoya (2021): Evaluation of epinephrine and vasopressin toxicity in human mesenchymal stem cells (hMSCs): in vitro study. Dissertation, LMU München: Faculty of Medicine
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Abstract

Purpose: Epinephrine is an effective vasoconstrictor, and it is commonly administered intra-articular during arthroscopy to control the bleeding as a local hemostatic agent. However, no study so far has investigated possible toxicities of either epinephrine or an alternative like vasopressin on human mesenchymal stem cells. Methods: Triple-layer culture flasks have been used to seed the hMSCs; we have seeded the cells at a density of 10,000 cells/cm2. The hMSCs were plated into 96-well plates for evaluation of mitochondrial activity or 6-well plates for evaluation of apoptosis, and all the plates contained culture medium. Cells were treated with either epinephrine or vasopressin, and the control group was untreated. Epinephrine and vasopressin were removed after forty minutes, and freshly prepared growth medium has been provided to the cells. We evaluated two apoptosis markers (CASP3 and PARP-1) and the activity of the mitochondria at one hour, twenty-four hours, and seven days. Results: Mitochondrial activity did not significantly decrease after exposure to either epinephrine or vasopressin. In the cells exposed to epinephrine, we noted a significant increase in PARP-1 level at seven days; we noted the same increase after exposure to vasopressin at twenty-four hours and seven days. The amount of CASP3 has significantly increased only at twenty-four hours in those cells which were exposed to epinephrine. Conclusions: These data suggest that neither epinephrine nor vasopressin influence mitochondrial activity in hMSCs in vitro. We observed an increase in apoptosis markers without a corresponding decrease in mitochondrial activity, which could indicate that both medications are toxic to hMSCs. The absence of a response of CASP3 after exposure to the vasopressin, suggest that vasopressin may be less toxic for hMSCs. However, while epinephrine and vasopressin are effective as hemostatic agents, they may have toxic effects on hMSCs, warranting caution in their use during arthroscopy, especially if performing the kind of procedure to induce the tissue repair and regeneration (for example, meniscal-repair, microfracture, tendon-repair, OATS, ACI, etc.).