Jalilzadeh-Tabrizi Sepideh, Pilehvar-Soltanahmadi Younes, Alizadeh Effat, Alipour Shahriar, Dadashpour Mehdi, Nejati-Koshki Kazem, Zarghami Nosratollah
1 Stem Cell Research Center, Tabriz University of Medical Sciences , Tabriz, Iran .
2 Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences , Tabriz, Iran .
Biopreserv Biobank. 2018 Apr;16(2):66-76. doi: 10.1089/bio.2017.0056. Epub 2018 Jan 22.
Emu oil (EO) with anti-inflammatory, antioxidative, and wound healing properties can be blended for preparing bioactive nanofibrous scaffold. Adipose tissue-derived stem cells (ADSCs) are promising candidates for tissue engineering, and preserving their stemness potential is vital for further therapeutic applications.
The aim of this study was to fabricate EO-blended nanofiber and investigate its effect on proliferation, survival, and stemness preservation of ADSCs.
Pure EO composition was characterized using a gas chromatograph mass spectrometer. EO-PCL-polyethylene glycol (PEG) nanofibers were successfully fabricated using an electrospinning technique and characterized by field emission scanning electron microscopy (FE-SEM) and fourier-transform infrared spectroscopy (FTIR). Cell viability and adhesion were measured using the MTT assay and FE-SEM. Finally, quantitative PCR (qPCR) was used to quantify the expression level of cell cycle regulated genes and pluripotency-associated transcription factors.
Findings showed that 20% (w/w) of EO is the optimum oil content in the electrospun solution to achieve good morphology and ultrafine fibers. The relatively high optical densities and FE-SEM images indicated that EO highly supported cell adhesion and proliferation on the matrices. In addition, EO-PCL-PEG electrospun nanofibrous mats significantly upregulated the expression levels of cell cycle regulated genes (Cyclin D1, pRb, and P53) and stemness markers (Nanog, OCT-4, Rex-1, and Sox-2) than PCL-PEG nanofiber and tissue culture polystyrene in 7 and 14 days of cell culture.
These results demonstrate that the EO-blended nanofibrous mat can be used as a bioactive scaffold to support cell adhesion and proliferation while simultaneously maintaining the stemness of ADSCs.
鸸鹋油(EO)具有抗炎、抗氧化和伤口愈合特性,可用于制备生物活性纳米纤维支架。脂肪组织来源的干细胞(ADSCs)是组织工程中有前景的候选细胞,保持其干细胞潜能对于进一步的治疗应用至关重要。
本研究旨在制备含EO的纳米纤维并研究其对ADSCs增殖、存活和干细胞潜能保持的影响。
使用气相色谱 - 质谱仪对纯EO成分进行表征。采用静电纺丝技术成功制备了EO - 聚己内酯 - 聚乙二醇(PEG)纳米纤维,并用场发射扫描电子显微镜(FE - SEM)和傅里叶变换红外光谱(FTIR)进行表征。使用MTT法和FE - SEM测量细胞活力和粘附情况。最后,采用定量聚合酶链反应(qPCR)定量细胞周期调控基因和多能性相关转录因子的表达水平。
研究结果表明,在静电纺丝溶液中20%(w/w) 的EO是实现良好形态和超细纤维的最佳油含量。相对较高的光密度和FE - SEM图像表明,EO高度支持细胞在基质上的粘附和增殖。此外,在细胞培养7天和14天时,与PCL - PEG纳米纤维和组织培养聚苯乙烯相比,EO - PCL - PEG静电纺纳米纤维垫显著上调了细胞周期调控基因(细胞周期蛋白D1、磷酸化视网膜母细胞瘤蛋白和P53)和干细胞标志物(Nanog、OCT - 4、Rex - 1和Sox - 2)的表达水平。
这些结果表明,含EO的纳米纤维垫可作为生物活性支架,支持细胞粘附和增殖,同时保持ADSCs的干细胞特性。