Department of Dentistry, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Department of Dentistry, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
J Cell Mol Med. 2020 May;24(9):5146-5151. doi: 10.1111/jcmm.15158. Epub 2020 Mar 23.
Periodontal ligament stem cells (PDLSCs) from beagle dogs had the characteristics of multi-directional differentiation and had great application potential in tissue engineering and cell regenerative medicine. In this study, we analysed the odontogenesis and neuronal differentiation characteristics of PDLSCs in vitro. Results showed that the calcined tooth powder (CTP) and silver nanoparticles (AgNPs) additives could induce the PDLSCs into odontogenesis differentiation; besides, the immunofluorescence staining identified that the high dosage calcined tooth powder (400 μg/mL) significantly facilitated the odontogenesis associated with BMP4 expression. While the nutritional factor (L-glutamine, NGF (nerve growth factor), bFGF (basic fibroblast growth factor), IGF-1 (insulin-like growth factor-1) and EGF (epidermal growth factor)) additives were prior to induce the PDLSCs into neuronal differentiation. Simultaneously, PDLSCs had high proliferation ability with the different supplemented additives. Importantly, the Western blot results also proved the BMP4 and SMAD1 proteins were highly expressed in the induced odontoblast, while the SOX1, NCAM1, GFAP and VEGFA proteins were all obviously expressed in the induced neurons. Hence, PDLSCs had characteristics of both odontogenesis and neuronal differentiation.
犬牙周韧带干细胞(PDLSCs)具有多向分化的特性,在组织工程和细胞再生医学中有很大的应用潜力。本研究分析了犬牙周韧带干细胞在体外的成牙和向神经元分化的特性。结果表明,煅烧牙粉(CTP)和银纳米颗粒(AgNPs)添加剂可诱导 PDLSCs 向成牙分化;此外,免疫荧光染色鉴定表明,高剂量煅烧牙粉(400μg/mL)显著促进了与 BMP4 表达相关的成牙。而营养因子(L-谷氨酰胺、神经生长因子(NGF)、碱性成纤维细胞生长因子(bFGF)、胰岛素样生长因子-1(IGF-1)和表皮生长因子(EGF))添加剂则优先诱导 PDLSCs 向神经元分化。同时,PDLSCs 具有较高的增殖能力,且不同的补充添加剂对其增殖能力的影响也不同。重要的是,Western blot 结果也证明了诱导的成牙细胞中 BMP4 和 SMAD1 蛋白的高表达,而诱导的神经元中 SOX1、NCAM1、GFAP 和 VEGFA 蛋白均明显表达。因此,PDLSCs 具有成牙和向神经元分化的特性。