Department of Biochemistry, Faculty of Basic Sciences, Islamic Azad University Damghan Branch, Damghan, Iran.
Department of Pilot Nanobiotechnology, Pasteur Institute of Iran, Tehran, Iran.
Int J Biol Macromol. 2020 May 1;150:152-160. doi: 10.1016/j.ijbiomac.2020.02.036. Epub 2020 Feb 6.
In this study, carboxymethyl chitosan (CMC) was grafted on polycaprolactone (PCL) nanofibers to fabricate scaffolds for bone tissue engineering. The electrospun PCL nanofibers were treated by cold atmospheric plasma (CAP) of helium to generate the reactive functions necessary for CMC grafting. β-carotene (βC) as a biochemical clue and electromagnetic field (EMF, 31.4 μT, 1 h per day) as a biophysical stimulator were used to promote the proliferation and osteodifferentiation of adipose mesenchymal stem cells (ADSCs). Alizarin red staining and calcium content results indicated the generation of nodal calcium on the CMC30%-g-PCL scaffold after 14 days of incubation in the presence or absence of external stimulation factors. Immunocytochemistry (ICC) results confirmed the expression of osteonectin protein for the stem cells seeded on CMC30%-g-PCL with or without using βC or EMF. These results suggest that the fabricated CMC-grafted scaffolds have the ability to self-differentiate stem cells to osteoblasts due to the osteoinductive effects of the grafted CMC. Furthermore, the osteodifferentiation of ADSCs is promoted by using an external stimulation factor such as βC or EMF.
在这项研究中，羧甲基壳聚糖（CMC）接枝到聚己内酯（PCL）纳米纤维上，以制备用于骨组织工程的支架。通过冷等离体（CAP）氦气处理电纺的 PCL 纳米纤维，以产生 CMC 接枝所需的反应性官能团。β-胡萝卜素（βC）作为生化线索和电磁场（EMF，31.4 μT，每天 1 小时）作为生物物理刺激物，用于促进脂肪间充质干细胞（ADSCs）的增殖和成骨分化。茜素红染色和钙含量结果表明，在存在或不存在外部刺激因素的情况下，在孵育 14 天后，在 CMC30%-g-PCL 支架上生成了节点钙。免疫细胞化学（ICC）结果证实了在有或没有使用 βC 或 EMF 的情况下，接枝 CMC 的 CMC30%-g-PCL 上接种的干细胞表达骨连蛋白蛋白。这些结果表明，由于接枝 CMC 的骨诱导作用，制备的接枝 CMC 的支架具有将干细胞自行分化为成骨细胞的能力。此外，通过使用βC 或 EMF 等外部刺激因子促进 ADSCs 的成骨分化。
