Evaluation of the osteogenic potential of rat adipose-derived stem cells with different polycaprolactone/alginate-based nanofibrous scaffolds: an in vitro study

使用不同的聚己内酯/海藻酸盐基纳米纤维支架评估大鼠脂肪干细胞的成骨潜能:一项体外研究

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作者:Eman Hany, Sarah Yahia, Mahmoud Fathy Elsherbeny, Nagla Mahmoud Salama, Islam Mohammed Ateia, Noha Tharwat Abou El-Khier, Ibrahim El-Sherbiny, Mazen Tharwat Abou Elkhier

Background

Bone tissue engineering is a widely growing field that requires the combination of cells, scaffolds and signaling molecules. Adipose derived stem cells (ADSCs) are an accessible and abundant source of mesenchymal stem cells with high plasticity. Polycaprolactone/alginate (PCL/Alg) composite scaffolds have been used in bone regeneration and nano-hydroxyapatite (n-HA) is used as a reinforcing, osteoconductive component in scaffold fabrication. This study was conducted to assess the ability of three different PCL/Alg based scaffolds to induce osteogenic differentiation of ADSCs and to compare between them.

Conclusions

ADSCs and PCL/Alg-based scaffolds compose a good tissue engineering complex for bone regeneration. Addition of n-HA to PCL/Alg scaffolds and crosslinking with CaCl2 efficiently improve the osteogenic potential of ADSCs.

Methods

The study comprised 5 groups; negative control group with ADSCs cultured in complete culture media, positive control group with ADSCs cultured in osteogenic differentiation media, and 3 experimental groups with ADSCs seeded onto 3 scaffolds: S1 (PCL/Alg), S2 (PCL/Alg/Ca) and S3 (PCL/Alg/Ca/n-HA) respectively and cultured in osteogenic media. Mineralization and gene expression were assessed by Alizarin red S (ARS) staining and real time quantitative polymerase chain reaction (RT-qPCR). Evaluation was done at 7, 14 and 21 days.

Results

ARS staining reflected a time dependent increase through days 7, 14 and 21, with S3 (PCL/Alg/Ca/n-HA) group showing the highest mineralization levels. RT-qPCR detected upregulation of ALP gene expression at day 7 and decline thereafter. S2 (PCL/Alg/Ca) and S3 (PCL/Alg/Ca/n-HA) groups showed significantly higher gene expression levels than S1 (PCL/Alg). Conclusions: ADSCs and PCL/Alg-based scaffolds compose a good tissue engineering complex for bone regeneration. Addition of n-HA to PCL/Alg scaffolds and crosslinking with CaCl2 efficiently improve the osteogenic potential of ADSCs.

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