Abstract
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.
Highlights
With the rise of tissue engineering, the construction of biological substitutes such as biological tissues, organs and materials, which were used to rescue or/restore various forms of damaged or defective human tissues during recent years, has become a hotspot in the research of tissues engineering.[1,2] In endodontic treatment, there were shortcomings of the conventional pulp capping material, calcium hydroxide[3]; the dental stem cells isolated from teeth and nearby tissues had been attracted more and more attention in investigating their utilities in dental prosthesis by researchers due to their self-renewal and multilineage differentiation potential.[4]
We had provided the evidence that the calcined tooth powder (CTP) and silver nanoparticles (AgNPs) additives had the ability to induce the Periodontal ligament stem cells (PDLSCs) differentiate into odontogenesis, while the nutritional factor (L-glutamine, neuron growth factor 1 (NGF), bFGF, insulin-like growth factor 1 (IGF-1) and epidermal growth factor (EGF)) additives were prior to induce the PDLSCs into neuronal differentiation
The CTP ingredients were major tooth calcium powder and hyaluronic acid, which were suitable for the migration and osteogenic differentiation of dental stem cells[17]; besides, its’ good biological compatibility made it to be a useful therapy for dental regeneration.[18]
Summary
With the rise of tissue engineering, the construction of biological substitutes such as biological tissues, organs and materials, which were used to rescue or/restore various forms of damaged or defective human tissues during recent years, has become a hotspot in the research of tissues engineering.[1,2] In endodontic treatment, there were shortcomings of the conventional pulp capping material, calcium hydroxide[3]; the dental stem cells isolated from teeth and nearby tissues had been attracted more and more attention in investigating their utilities in dental prosthesis by researchers due to their self-renewal and multilineage differentiation potential.[4]. | 5147 periodontal tissues.[5,6] While the PDLSCs were obtained from the beagle dog periodontal ligament (PDL) of teeth freshly extracted, which had been recognized as a useful cell source for periodontal tissue regeneration.[7] They had the characteristics of easy to access and cryopreserved conveniently; besides, studies identified that they could induce the formation of new blood vessels and improve osteogenic.[8,9] In vitro culture studies, it had been reported that they could differentiate into odontoblasts, osteoblasts, chondrocytes and neurons 10-12; the PDLSCs had great utility in regenerative medicine. The cells were washed twice with PBS; subsequently, the nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI; 1:1000, Sigma) for 1 hour at 4°C, washed them with PBS again and mounted the cell slide with antifade mounting medium; at last, the images of the coverslips were visualized with an inverted fluorescence microscope (Olympus)
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