Outline

  • Abstract
  • Graphical Abstract
  • Abbreviations
  • Keywords
  • 1. Introduction
  • 2. Materials and Methods
  • 2.1. Study Design and Setting
  • 2.2. Adscs Isolation
  • 2.3. Adscs Characterization
  • 2.4. Adscs Culture and Drug-Treatment
  • 2.5. Adscs Viability Assay
  • 2.6. Real-Time Pcr
  • 2.7. Oil Red O Staining and Quantification
  • 2.8. Alizarin Red S Staining and Quantification
  • 2.9. Statistical Analysis
  • 3. Results
  • 3.1. Isolation and Characterization of Adscs
  • 3.2. Effects of Drug-Treatment on Stem Cell Proliferation and Stemness Properties
  • 3.3. Effects of Drug-Treatment on Adipogenic Commitment
  • 3.4. Effects of Drug-Treatment on Osteogenic Commitment
  • 4. Discussion
  • 5. Conclusion
  • Conflict of Interest
  • Funding
  • Acknowledgements
  • References

رئوس مطالب

  • خلاصه
  • کلیدواژه ها
  • 1. مقدمه
  • 2. مواد و روش ها
  • 2.1. طراحی مطالعه
  • 2.2.جداسازی ADSC ها
  • 2.3. ویژگی ADSC ها
  • 2.4. کشت ADSC ها و مواجه دارویی
  • 2.5. ارزیابی زنده بودن سلول های ADSC
  • 2.6. ریم تایم PCR
  • 2.7. رنگ آمیزی Oil Red O و کمی سازی
  • 2.8. رنگ آمیزی آلیزارین رد S و کمی سازی
  • 2.9.تحلیل آماری
  • 3. نتایج
  • 3.1. جداسازی و ویژگی ADSC ها
  • 3.2. تاثیرات مواجهه دارویی روی تکثیر سلول بنیادی و ویژگیهای بنیادی
  • 3.3. تاثیرات مواجهه دارو بر روی تمایز ادیپوژنیک
  • 3.4. تاثیرات مواجه دارویی بر روی تمایز استئوژنیک
  • 4. بحث
  • 5. نتیجه گیری

Abstract

It is known that users of psychotropic drugs often have weight gain, adverse effects on bone mineral density and osteoporosis, but the molecular basis for these side effects is poorly understood. The aim of this study is to evaluate the effects in vitro of duloxetine (a serotonin and norepinephrine reuptake inhibitor) and fluoxetine (a selective serotonin reuptake inhibitor) on the physiology of human adult stem cells. Adipose-derived stem cells (ADSCs) were isolated and characterized investigating phenotype morphology, expression and frequency of surface markers. Then, a non-toxic concentration of duloxetine and fluoxetine was selected to treat cells during adipogenic and osteogenic differentiation. Stemness properties and the differentiation potential of drug-treated cells were investigated by the quantification of adipogenic and osteogenic markers gene expression and histological staining. The collected data showed that the administration of a daily non-toxic dose of duloxetine and fluoxetine has not directly influenced ADSCs proliferation and their stemness properties. The treatment with duloxetine or fluoxetine did not lead to morphological alterations during adipogenic or osteogenic commitment. However, treatments with the antidepressant showed a slight difference in adipogenic gene expression timing. Furthermore, duloxetine treatment caused an advance in gene expression of early and late osteogenic markers. Fluoxetine instead caused an increase in expression of osteogenic genes compared to untreated cells. In contrast, in pre-differentiated cells, the daily treatment with duloxetine or fluoxetine did not alter the expression profile of adipogenic and osteogenic differentiation. In conclusion, a non-toxic concentration of duloxetine and fluoxetine does not alter the stemness properties of ADSCs and does not prevent the commitment of pre-differentiated ADSCs in adipocytes or osteocyte. Probably, the weight gain and osteoporotic effects associated with the use of psychotropic drugs could be closely related to the direct action of serotonin.

Keywords: - - -

Conclusions

In conclusion, these results demonstrate that a non-toxic concentration of duloxetine or fluoxetine have time-dependent effects on mesenchymal stem cells. In particular, they influence proliferation and stemness properties of non-committed ADSCs in short term, indeed, after 21 days of daily drug-treatments both cell proliferation and mesenchymal stromal cell markers expression are equal to the control condition. The treatment with fluoxetine do not lead to morphological alterations during adipogenic or osteogenic commitment while the one with duloxetine seems to decrease lipid accumulation and to increase mineral deposition.

Nevertheless, both drug treatments influence the gene expression timing of adipogenic genes in committed cells. Instead, in osteogenic commitment, duloxetine determines an anticipation of early and late osteogenic markers gene expression, and fluoxetine causes a significant increase in osteogenic genes expression. Our study suggest that, despite affecting mesenchymal stem cells commitment, the effects of duloxetine and fluoxetine on body weight and bone metabolism probably not correlate directly with mesenchymal stem cells differentiation; perhaps the effects observed on patients could be closely related to the action of the drugs on serotonin reuptake. For these reason further studies that deeply investigate the mechanism related to antidepressant side effects will be necessary.

Since the use of antidepressants in clinical practice has dramatically increased in recent years, understanding how these drugs are associated with weight gain, osteoporosis and fracture risk is pivotal and this may influence prescribing practices as knowledge increases.

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