مقالات پذیرفته شده در نهمین کنگره بین المللی زیست پزشکی
Evaluation of OSTEONECTIN Protein Expression in Mesenchymal Stem Cell Encapsulated into Chitosan/Carboxymethyl chitosan Hydrogel Treated with Exosome
Evaluation of OSTEONECTIN Protein Expression in Mesenchymal Stem Cell Encapsulated into Chitosan/Carboxymethyl chitosan Hydrogel Treated with Exosome
Rozhin Rajabi,1Shiva Irani,2Fereshteh Sharifi,3,*Marjan Mohammadali,4
1. Department of Biology, SR.C., Islamic Azad University, Tehran, Iran 2. Department of Biology, SR.C., Islamic Azad University, Tehran, Iran 3. Department of Biology, CT.C., Islamic Azad University, Tehran, Iran 4. Department of Biology, SR.C., Islamic Azad University, Tehran, Iran
Introduction: Extracellular nanovesicles secreted by mesenchymal stem cells have been introduced as novel bioactive agents for inducing cell growth and osteogenic differentiation. The synergistic effect of mesenchymal-derived exosomes (Exo) with Chitosan/Carboxymethyl chitosan hydrogel (CS/CMC) can enhance scaffold performance and promote proliferation and osteogenic differentiation of adipose-derived mesenchymal stem cells (AMSCs). The purpose of this study was to investigate the impact of Exo on the biocompatibility, proliferation, and osteogenic differentiation of AMSCs encapsulated in CS/CMC hydrogel.
Methods: The morphology of CS/CMC hydrogel scaffolds was examined using scanning electron microscopy (SEM). To evaluate biocompatibility, AMSCs were cultured in CS/CMC hydrogel for 24 hours and observed using an inverted microscope. Cell viability and proliferation of encapsulated AMSCs in CS/CMC hydrogel along with and without Exo were evaluated by MTT assay at 24 hours and on day 7. The osteogenic differentiation potential of AMSCs encapsulated in CS/CMC hydrogel treated with Exo and in CS/CMC hydrogel was assessed on day 7 using the Calcium Content Assay to quantitatively evaluate calcium levels, and immunocytochemistry (ICC) with DAPI to examine OSTEONECTIN protein expression and confirm the genetic integrity and viability of the cells.
Results: SEM analysis showed that the CS/CMC hydrogel exhibited a porous structure. Images of AMSCs cultured in CS/CMC hydrogel for 24 hours demonstrated the biocompatibility of the hydrogel. The MTT assay revealed a significant increase in viability and proliferation of encapsulated AMSCs in CS/CMC-Exo hydrogel at 24 hours and on day 7 (p < 0.05). Calcium content analysis showed that calcium deposition in AMSCs encapsulated in CS/CMC hydrogel treated with Exo significantly increased on day 7 (p < 0.05). ICC analysis of AMSCs encapsulated in CS/CMC hydrogel treated with Exo, performed with DAPI, confirmed OSTEONECTIN protein expression and verified the genetic integrity and viability of the cells (39.61%).
Conclusion: Incorporation of Exo into CS/CMC hydrogels enhances scaffold biocompatibility, promotes proliferation of encapsulated AMSCs, and stimulates their osteogenic differentiation. These findings suggest that CS/CMC hydrogels enriched with Exo have great potential as effective scaffolds for bone regeneration.