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Investigation of the Synergism Between Engineered Stem Cells with Vtk Containing Construct and Electrical Stimulation in Osteogenic Differentiation

Gharaei, Mohammad Saleh | 2023

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 57092 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Alemzadeh, Iran; Bakhshandeh, Behnaz; Saadatmand, Maryam
  7. Abstract:
  8. In recent years, advances in medical science and increasing the level of healthcare have increased the life expectancy index; But in the meantime, a phenomenon called “diseases related to life expectancy” has appeared. Bone and skeletal diseases have a significant share in this category because the passage of time reduces bone mineral density and this is the source of many diseases, including osteoporosis. Current bone therapies, which include autologous and allogeneic bone grafts, will not meet this high need. Bone tissue engineering has been proposed as a potential alternative to solve this challenge. In bone tissue engineering, the essential components required for bone formation and development include appropriate cells, biocompatible scaffolds, growth factors, and vascularization to supply nutrients to the growing tissue. Usually, the cells used in tissue engineering are stem cells, and the main challenge faced by this department is the optimal differentiation of stem cells into osteoprogenitor cells and then into osteoblasts. Electrical stimulation of cells and the use of bioactive peptides are among the solutions that have been used in recent years to improve osteogenic differentiation. In this research, we tried to investigate the synergism of these two factors in osteogenic differentiation. Polycaprolactone scaffold was prepared by electrospinning and bone marrow mesenchymal stem cells (BMSCs) were selected and harvested. Studying the image of the cells under a scanning electron microscope (SEM) showed that the cells were well placed on the scaffold. The VTK-DPI peptide was transfected into the cell using lipofectamine 2000 and the cells were electrically stimulated using a metal electrode and conditions of 100 Hz frequency and 2 V voltage. Cells were evaluated on days 2 and 5 using calcium and alkaline phosphatase biochemical analysis. Studying and comparison of the experimental groups showed that the transfected groups recorded a lower calcium index on day 2 due to the shock caused by transfection, but the index of this group made a remarkable change on day 5 and showed a significant difference with non-transfected groups. In total, the group that underwent both electrical induction and peptide transfection recorded the highest calcium level. Peptide transfection caused calcium secretion to occur earlier, but it had no effect on alkaline phosphatase. So, the amount of alkaline phosphatase in different groups was almost the same on days 2 and 5, which is due to the fact that the peak secretion of alkaline phosphatase occurs on days 7-10 after Seeding. Overall, the results showed that the combination and synergy of electrical stimulation and peptide transfection is a positive factor for osteogenic differentiation
  9. Keywords:
  10. Bone Tissue Engineering ; Electrical Stimulation ; Stem Cell ; Engineered Stem Cell ; Osteogenic Differentiation ; VTK-DPI Peptide

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