Hany Eman, Yahia Sarah, Elsherbeny Mahmoud Fathy, Salama Nagla Mahmoud, Ateia Islam Mohammed, Abou El-Khier Noha Tharwat, El-Sherbiny Ibrahim, Abou Elkhier Mazen Tharwat
Oral Biology Department, Faculty of Dentistry, Mansoura University, Mansoura, Egypt.
Nanomedicine Lab, Center of Material Science, University of Science and Technology, Zewail City of Science and Technology, Giza, Egypt.
Stem Cell Investig. 2020 Aug 7;7:14. doi: 10.21037/sci-2020-015. eCollection 2020.
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.
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.
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 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).
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.
骨组织工程是一个发展迅速的领域,需要将细胞、支架和信号分子相结合。脂肪来源干细胞(ADSCs)是一种易于获取且丰富的间充质干细胞来源,具有高度可塑性。聚己内酯/海藻酸盐(PCL/Alg)复合支架已用于骨再生,纳米羟基磷灰石(n-HA)用作支架制造中的增强、骨传导成分。本研究旨在评估三种不同的基于PCL/Alg的支架诱导ADSCs成骨分化的能力并进行比较。
本研究包括5组;阴性对照组为ADSCs在完全培养基中培养,阳性对照组为ADSCs在成骨分化培养基中培养,3个实验组为ADSCs分别接种到3种支架上:S1(PCL/Alg)、S2(PCL/Alg/Ca)和S3(PCL/Alg/Ca/n-HA),并在成骨培养基中培养。通过茜素红S(ARS)染色和实时定量聚合酶链反应(RT-qPCR)评估矿化和基因表达。在第7、14和21天进行评估。
ARS染色显示在第7、14和21天随时间增加,S3(PCL/Alg/Ca/n-HA)组显示出最高的矿化水平。RT-qPCR检测到第7天基因表达上调,此后下降。S2(PCL/Alg/Ca)和S3(PCL/Alg/Ca/n-HA)组的基因表达水平显著高于S1(PCL/Alg)组。
ADSCs和基于PCL/Alg的支架构成了用于骨再生的良好组织工程复合物。向PCL/Alg支架中添加n-HA并与CaCl2交联可有效提高ADSCs的成骨潜力。