Dental and Periodontal Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Faculty of Dentistry, Tabriz University of Medical Sciences, Tabriz, Iran.
Biomed Res Int. 2022 Feb 14;2022:5401461. doi: 10.1155/2022/5401461. eCollection 2022.
Chitosan is a natural biopolymer that attracted enormous attention in biomedical fields. The main components of regenerative endodontic procedures (REPs), as well as tissue engineering, are scaffolds, stem cells, and growth factors. As one of the basic factors in the REPs is maintaining vascularization, this study was aimed at developing basic fibroblast growth factor- (bFGF-) loaded scaffolds and investigating their effects on the angiogenic induction in human dental pulp stem cells (hDPSCs).
Poly (-caprolactone) (PCL)/chitosan- (CS-) based highly porous scaffold (PCL/CS) was prepared and evaluated by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) analyses. The adhesion and survival potency of seeded cells were assessed by SEM and MTT assays, respectively. The amount of angiogenic markers was investigated in gene and protein levels by real-time PCR and western blotting assays, respectively.
Based on our findings, the SEM and FTIR tests confirmed the appropriate structure of synthesized scaffolds. Besides, the adhesion and survival rate of cells and the levels of VEGFR-2, Tie2, and Angiopoietin-1 genes were increased significantly in the PCL/CS/bFGF group. Also, the western blotting results showed the upregulation of these markers at protein levels, which were considerably higher at the PCL/CS/bFGF group ( < 0.05).
On a more general note, this study demonstrates that the bFGF-loaded PCL/CS scaffolds have the potential to promote angiogenesis of hDPSCs, which could provide vitality of dentin-pulp complex as the initial required factor for regenerative endodontic procedures.
壳聚糖是一种天然的生物聚合物,在生物医学领域引起了极大的关注。再生牙髓治疗(REPs)以及组织工程的主要成分是支架、干细胞和生长因子。由于 REPs 的基本因素之一是保持血管化,因此本研究旨在开发碱性成纤维细胞生长因子(bFGF)负载的支架,并研究其对人牙髓干细胞(hDPSCs)血管生成诱导的影响。
聚(-己内酯)(PCL)/壳聚糖(CS)基高多孔支架(PCL/CS)通过扫描电子显微镜(SEM)和傅里叶变换红外光谱(FTIR)分析进行了评估。通过 SEM 和 MTT 分析分别评估了接种细胞的粘附和存活能力。通过实时 PCR 和 Western blot 分析分别在基因和蛋白水平上研究了血管生成标志物的含量。
根据我们的发现,SEM 和 FTIR 测试证实了合成支架的合适结构。此外,PCL/CS/bFGF 组细胞的粘附和存活率以及 VEGFR-2、Tie2 和血管生成素-1 基因的水平显著增加。此外,Western blot 结果表明这些标志物在蛋白水平上的上调,PCL/CS/bFGF 组的上调更为明显(<0.05)。
总的来说,本研究表明,负载 bFGF 的 PCL/CS 支架具有促进 hDPSCs 血管生成的潜力,这可以为再生牙髓治疗提供牙本质-牙髓复合体的活力,作为再生牙髓治疗的初始必需因素。