Del Angel-Mosqueda Casiano, Gutiérrez-Puente Yolanda, López-Lozano Ada Pricila, Romero-Zavaleta Ricardo Emmanuel, Mendiola-Jiménez Andrés, Medina-De la Garza Carlos Eduardo, Márquez-M Marcela, De la Garza-Ramos Myriam Angélica
Unidad de Odontología Integral y Especialidades, Centro de Investigación y Desarrollo en Ciencias de la Salud, Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, México.
Instituto de Biotecnología, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Nuevo León, México.
Head Face Med. 2015 Sep 3;11:29. doi: 10.1186/s13005-015-0086-5.
Epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) play an important role in extracellular matrix mineralization, a complex process required for proper bone regeneration, one of the biggest challenges in dentistry. The purpose of this study was to evaluate the osteogenic potential of EGF and bFGF on dental pulp stem cells (DPSCs).
Human DPSCs were isolated using CD105 magnetic microbeads and characterized by flow cytometry. To induce osteoblast differentiation, the cells were cultured in osteogenic medium supplemented with EGF or bFGF at a low concentration. Cell morphology and expression of CD146 and CD10 surface markers were analyzed using fluorescence microscopy. To measure mineralization, an alizarin red S assay was performed and typical markers of osteoblastic phenotype were evaluated by RT-PCR.
EGF treatment induced morphological changes and suppression of CD146 and CD10 markers. Additionally, the cells were capable of producing calcium deposits and increasing the mRNA expression to alkaline phosphatase (ALP) and osteocalcin (OCN) in relation to control groups (p < 0.001). However, bFGF treatment showed an inhibitory effect.
These data suggests that DPSCs in combination with EGF could be an effective stem cell-based therapy for bone tissue engineering applications in periodontics and oral implantology.
表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF)在细胞外基质矿化过程中发挥着重要作用,细胞外基质矿化是骨组织正常再生所需的一个复杂过程,而骨组织再生是牙科领域面临的最大挑战之一。本研究的目的是评估EGF和bFGF对牙髓干细胞(DPSCs)的成骨潜能。
使用CD105磁性微珠分离人牙髓干细胞,并通过流式细胞术进行鉴定。为诱导成骨细胞分化,将细胞在添加低浓度EGF或bFGF的成骨培养基中培养。使用荧光显微镜分析细胞形态以及CD146和CD10表面标志物的表达。为测量矿化情况,进行茜素红S测定,并通过逆转录聚合酶链反应(RT-PCR)评估成骨细胞表型的典型标志物。
EGF处理诱导了细胞形态变化以及CD146和CD10标志物的表达抑制。此外,与对照组相比,这些细胞能够产生钙沉积,并增加碱性磷酸酶(ALP)和骨钙素(OCN)的mRNA表达(p < 0.001)。然而,bFGF处理显示出抑制作用。
这些数据表明,DPSCs与EGF联合使用可能是一种有效的基于干细胞的治疗方法,可用于牙周病学和口腔种植学中的骨组织工程应用。