مقالات پذیرفته شده در نهمین کنگره بین المللی زیست پزشکی
Study of synergize of the chitosan hydrogel in conditioned media on differentiation of Mesenchymal stem cells for skin tissue engineering
Study of synergize of the chitosan hydrogel in conditioned media on differentiation of Mesenchymal stem cells for skin tissue engineering
Shiva Hajimousaei,1Shiva Irani,2Fereshteh Sharifi,3Seyed Mohammad Atyabi,4,*Marjan Mohamadali,5
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. Deparment of Nanobiotechnology Pasteur Institute of Iran, Tehran, Iran 5. Deparment of Nanobiotechnology Pasteur Institute of Iran, Tehran, Iran
Introduction: Nowadays skin injuries remain a big challenge in regenerative medicine. Over the past years among different structures, hydrogels have gained a lot of attention due to specific scaffold properties. They can mimic the extracellular matrix of native tissue and provide a supportive environment for cell attachment, consequently promoting cell proliferation and differentiation. Therefore the use of hydrogel scaffolds has a great potential in skin tissue engineering. Chitosan (CTS) is a natural hydrogel that stands out because it is biocompatible, biodegradable, and relatively easy to work with.
On the other hand, conditioned media (CM), a complex repertoire of bioactive molecules secreted by stem cells, can initiate a chain of signaling events leading to wound healing. The main aim of this study was to investigate whether the present of CTS hydrogel in CM could promote the differentiation of adipose-derived mesenchymal stem cells (AMSCs). To assess this impact, we focused particularly on the expression of collagen type I, since it represents the predominant extracellular matrix protein synthesized by fibroblasts and serves as a critical marker of their functional activity in tissue repair.
Methods: First, we synthesized the CTS hydrogel and characterized with Scanning Electron Microscopy (SEM). The CM was collected from mesenchymal stem cells that had been cultured for 24 hours. After that we cultured 10⁴ AMSCs in high-glucose DMEM medium and treated them with hydrogel. Then, we confirmed the morphology of cells using SEM in 72 hours. We performed an MTT assay over 24 hours. Actually it was for checking the viability and compatibility of the AMSCs. Finally, we carried out real-time PCR to measure the expression of collagen type I.
Results: For characterizing the CTS hydrogel, SEM image showed that the hydrogel displayed a suitable porous morphology. After 72 hour of treating cells with hydrogel, SEM image showed the interaction between them and hydrogel. It showed the cells are in good morphological condition. They are also attached to the hydrogel and the interaction of the cells with each other and the cells with the CTS hydrogel is clearly visible. The MTT assay showed that after 24 hours, the sample had a significant increase compared to the control sample (P=0.000). According to the results of the Real-time PCR tests, the differentiation markers of AMSCs in the in CM had a increase compared to the control (p < 0.001).
Conclusion: Our findings showed that the cells maintained good viability and morphology in the environment enriched with CM. The interaction between cells and the scaffold structure was evident, and the expression of collagen type I was increased. This indicates that CM can enhance AMSCs activity and promote their differentiation. Therefore, CM can be considered as an effective biological supplement for skin tissue engineering, as it not only improves cell health and attachment but also facilitates functional differentiation.