Khazaeel Kaveh, Sadeghi Abbas, Khademi Moghaddam Fatemeh, Mohammadi Tayebeh
Department of Basic Sciences, Division of Anatomy and Embryology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
Stem Cells and Transgenic Technology Research Center (STTRC), Shahid Chamran University of Ahvaz, Ahvaz, Iran.
Cytotechnology. 2025 Feb;77(1):14. doi: 10.1007/s10616-024-00672-9. Epub 2024 Dec 10.
Bone tissue engineering is a promising approach to overcome the limitations of traditional autograft bone transplantation. Graphene quantum dots (GQDs) have been suggested as an enhancement for osteogenic differentiation. This study aimed to investigate the ability of the fibrin hydrogel scaffold in the presence of graphene quantum dots to promote osteogenic differentiation of human Wharton's jelly-derived mesenchymal stem cells (hWJ-MSCs). The hWJ-MSCs were isolated from the Wharton's jelly of the human umbilical cord using a mechanical method. Fibrin hydrogel scaffolds were prepared by mixing 15 µl of thrombin solution with fibrinogen solution. GQDs were incorporated into the scaffolds at concentrations of 0, 5, and 10 µg/ml. Cell viability was determined through DAPI staining and the MTT assay. Osteogenic differentiation was assessed by measuring alkaline phosphatase (ALP) activity, quantifying calcium deposition using Alizarin Red S staining, and analyzing the gene expression of BGLAP, COL1A1, Runx-2 and ALP via qPCR. Scanning electron microscopy (SEM) was employed to analyze the scaffold architecture. SEM analysis revealed that the fibrin hydrogel exhibited a suitable architecture for tissue engineering, and DAPI staining confirmed cell viability. The MTT results indicated that the GQDs and fibrin hydrogel scaffold exhibited no cytotoxic effects. Furthermore, the incorporation of GQDs at a concentration of 10 µg/ml significantly enhanced ALP activity, calcium deposition, and the expression of osteogenesis-related genes compared to the control. The findings suggest that the combination of fibrin hydrogel and GQDs can effectively promote the osteogenic differentiation of hWJ-MSCs, contributing to the advancement of bone tissue engineering.
骨组织工程是一种克服传统自体骨移植局限性的有前景的方法。石墨烯量子点（GQDs）已被认为可增强成骨分化。本研究旨在探讨在存在石墨烯量子点的情况下，纤维蛋白水凝胶支架促进人脐带华通氏胶来源间充质干细胞（hWJ-MSCs）成骨分化的能力。hWJ-MSCs通过机械方法从人脐带的华通氏胶中分离出来。通过将15微升凝血酶溶液与纤维蛋白原溶液混合制备纤维蛋白水凝胶支架。GQDs以0、5和10微克/毫升的浓度掺入支架中。通过DAPI染色和MTT法测定细胞活力。通过测量碱性磷酸酶（ALP）活性、使用茜素红S染色定量钙沉积以及通过qPCR分析BGLAP、COL1A1、Runx-2和ALP的基因表达来评估成骨分化。采用扫描电子显微镜（SEM）分析支架结构。SEM分析表明纤维蛋白水凝胶展现出适合组织工程的结构，并且DAPI染色证实了细胞活力。MTT结果表明GQDs和纤维蛋白水凝胶支架没有细胞毒性作用。此外，与对照组相比，以10微克/毫升的浓度掺入GQDs显著增强了ALP活性、钙沉积以及成骨相关基因的表达。这些发现表明纤维蛋白水凝胶和GQDs的组合可有效促进hWJ-MSCs的成骨分化，有助于骨组织工程的发展。
