ﺳﺎﺧﺖ دارﺑﺴﺖ ﻓﯿﺒﺮوﺋﯿﻨﯽ اﻟﮑﺘﺮورﯾﺴﯽ ﺷﺪه و ﺗﺎﺛﯿﺮ ﭘﯿﺶ اﻧﮑﻮﺑﺎﺳﯿﻮن آن در ﻣﺤﯿﻂ ﮐﺸﺖ ﺑﺮ روي ﺑﻘﺎ و ﭼﺴﺒﻨﺪﮔﯽ ﺳﻠﻮل ﻫﺎي ﻣﺰاﻧﺸﯿﻤﯽ ﻣﻐﺰ اﺳﺘﺨﻮان رت

Fabrication of electrospun silk fibroin scaffold and the effect of its pre-incubation in culture medium on survival and adhesion of rat bone marrow mesenchymal cells

اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﺑﻪ ﻣﻨﻈﻮر ﺳﻨﺘﺰ دارﺑﺴﺖ ﻓﯿﺒﺮوﺋﯿﻨﯽ اﻟﮑﺘﺮورﯾﺴﯽ ﺷﺪه و ﺑﺮرﺳﯽ ﺗﺎﺛﯿﺮ واﺑﺴﺘﻪ ﺑﻪ زﻣﺎن ﭘﯿﺶ-اﻧﮑﻮﺑﺎﺳﯿﻮن آن در ﻣﺤﯿﻂ ﮐﺸﺖ ﺑﺮ روي ﭼﺴﺒﻨﺪﮔﯽ ﺳﻠﻮﻟﯽ و ﺗﮑﺜﯿﺮ ﺳﻠﻮل ﻫﺎي ﮐﺎﺷﺘﻪ ﺷﺪه ﺑﺮ روي دارﺑﺴﺖ اﻧﺠﺎم ﮔﺮدﯾﺪ. اﺑﺘﺪا ﺳﺮﯾﺴﯿﻦ زداﺋﯽ ﭘﯿﻠﻪ اﺑﺮﯾﺸﻢ و آﻣﺎده ﺳﺎزي ﻓﯿﺒﺮوﺋﯿﻦ اﻧﺠﺎم ﺷﺪ و ﺳﭙﺲ ﻣﺤﻠﻮل ﻓﯿﺒﺮوﺋﯿﻦ 3% وزن/ﺣﺠﻢ ﺑﺎ اﺳﺘﻔﺎده از اﺳﯿﺪ ﻓﺮﻣﯿﮏ ﺗﻬﯿﻪ ﺷﺪ. ﺳﭙﺲ ﺑﺎ دﺳﺘﮕﺎه اﻟﮑﺘﺮورﯾﺴﯽ آزﻣﺎﯾﺸﮕﺎﻫﯽ، دارﺑﺴﺖ ﻓﯿﺒﺮوﺋﯿﻨﯽ اﻟﮑﺘﺮورﯾﺴﯽ ﺷﺪه ﺳﺎﺧﺘﻪ ﺷﺪه و ﺑﺎ ﻣﯿﮑﺮوﺳﮑﻮپ اﻟﮑﺘﺮوﻧﯽ ﻧﮕﺎره ﻣﻮرد ارزﯾﺎﺑﯽ ﻗﺮار ﮔﺮﻓﺖ. ﭘﯿﺶ- اﻧﮑﻮﺑﺎﺳﯿﻮن دارﺑﺴﺖ در ﻣﺤﯿﻂ ﮐﺸﺖ ﺑﻪ ﻣﺪت ﺻﻔﺮ، 1، 6، و 10 روز اﻧﺠﺎم ﺷﺪ و ﺑﺎ روش اﻧﺪازه ﮔﯿﺮي زاوﯾﻪ ﺗﻤﺎس ﻗﻄﺮه آب، ﻣﯿﺰان آب دوﺳﺘﯽ دارﺑﺴﺖ ﻫﺎ ارزﯾﺎﺑﯽ ﮔﺮدﯾﺪ. ﺳﭙﺲ ﺳﻠﻮل ﻫﺎي ﻣﺰاﻧﺸﯿﻤﯽ ﻣﻐﺰ اﺳﺘﺨﻮان رت ﺟﺪاﺳﺎزي، ﮐﺸﺖ، و ﺑﺮ روي دارﺑﺴﺖ ﻫﺎ ﮐﺎﺷﺖ ﮔﺮدﯾﺪ. ﺑﻌﺪ از 21 روز از ﮐﺎﺷﺖ ﺳﻠﻮل ﻫﺎ، ﻣﯿﺰان ﺑﻘﺎ ﺳﻠﻮل ﻫﺎ )ﺑﻪ روش MTT( و ﻏﻠﻈﺖ DNA ژﻧﻮﻣﯽ ﺳﻠﻮل ﻫﺎي ﭼﺴﺒﯿﺪه ﺑﻪ دارﺑﺴﺖ ﻣﻮرد ارزﯾﺎﺑﯽ ﻗﺮار ﮔﺮﻓﺖ. ﻧﺘﺎﯾﺞ ﻧﺸﺎن دادﻧﺪ ﮐﻪ اﻓﺰاﯾﺶ ﻣﺪت زﻣﺎن ﭘﯿﺶ-اﻧﮑﻮﺑﺎﺳﯿﻮن دارﺑﺴﺖ در ﻣﺤﯿﻂ ﮐﺸﺖ ﻣﻨﺠﺮ ﺑﻪ ﮐﺎﻫﺶ زاوﯾﻪ ﺗﻤﺎس و اﻓﺰاﯾﺶ ﺑﻘﺎ و ﺗﮑﺜﯿﺮ ﺳﻠﻮل ﻫﺎ ﮔﺮدﯾﺪ. ﺑﻪ ﻃﻮر ﮐﻠﯽ ﻣﻄﺎﻟﻌﻪ ﮐﻨﻮﻧﯽ ﻧﺸﺎن داد ﮐﻪ ﭘﯿﺶ - اﻧﮑﻮﺑﺎﺳﯿﻮن دارﺑﺴﺖ ﻓﯿﺒﺮوﺋﯿﻨﯽ اﻟﮑﺘﺮورﯾﺴﯽ ﺷﺪه ﺑﺎ اﻻﺳﺘﯿﺴﯿﺘﻪ ﺛﺎﺑﺖ در ﻣﺤﯿﻂ ﮐﺸﺖ ﻣﻨﺠﺮ ﺑﻪ اﻓﺰاﯾﺶ آب دوﺳﺘﯽ دارﺑﺴﺖ و ﻣﺘﻌﺎﻗﺒﺎ اﻓﺰاﯾﺶ ﺗﮑﺜﯿﺮ و ﺑﻘﺎ ﺳﻠﻮل ﻫﺎي ﻣﺰاﻧﺸﯿﻤﯽ ﮐﺎﺷﺘﻪ ﺷﺪه ﺑﺮ روي آن ﻣﯽ ﮔﺮدد. از اﯾﻦ ﯾﺎﻓﺘﻪ ﻣﯽ ﺗﻮان ﺑﻪ ﻋﻨﻮان ﯾﮏ ﻓﺎﮐﺘﻮر ﻣﻮﺛﺮ در آﻣﺎده ﺳﺎزي ﮐﺎﺷﺖ ﺳﻠﻮل ﻫﺎي ﻏﻀﺮوﻓﯽ ﺑﺮ روي دارﺑﺴﺖ در ﻣﻬﻨﺪﺳﯽ ﺑﺎﻓﺖ اﺳﺘﻔﺎده ﮐﺮد.

This study was performed to synthesize electrospun silk fibroin scaffold and to investigate the time-dependent effect of its pre-incubation in culture medium on the adhesion and proliferation of cells seeded on the scaffold. sericin was removed from silk cocoon and fibrin solution (3% w/v) was prepared using formic acid. Then, electrospun silk fibroin scaffold was made using lab-scale electrospinning machine and evaluated by scanning electron microscopy. Pre-incubation of scaffolds in culture medium was performed for 0, 1, 6, and 10 days and the hydrophilicity of scaffolds were evaluated by measuring the water contact angle. Rat bone marrow mesenchymal cells were then isolated, cultured, and seeded on scaffolds. After 21 days of cell seeding, cell viability (MTT method) and genomic DNA concentration of cells attached to the scaffold were evaluated. The results showed that increasing the pre-incubation time in the culture medium decreased the water contact angle and increased the survival and proliferation of cells. In general, the present study showed that pre-incubation of electrospun fibroin scaffold with constant elasticity in culture medium leads to increased scaffold hydrophilicity and consequently increases the proliferation and survival of mesenchymal cells seeded on it. Conclusion: these findings can be used as an effective factor in seeding cartilage cell on scaffolds in tissue engineering.