Lei Zhu1, Juan Ma2, Rui Mu2, Ruiqiao Zhu2, Feng Chen1, Xiaocui Wei2, Xiaolei Shi2, Shengqi Zang3, Lei Jin4. 1. Department of Stomatology, Jinling Hospital, Jinling Medical School of Nanjing University, Medical School of Southern Medical University, Clinic School of Nanjing Medical University, Clinic School of the Fourth Military Medical University, Nanjing, Jiangsu 210002, People's Republic of China; State Key laboratory of Military Stomatology, Department of Prosthodontics, School of Stomatology, The Fourth Military Medical University, Xian, Shaanxi 710023, People's Republic of China. 2. Department of Stomatology, Jinling Hospital, Jinling Medical School of Nanjing University, Medical School of Southern Medical University, Clinic School of Nanjing Medical University, Clinic School of the Fourth Military Medical University, Nanjing, Jiangsu 210002, People's Republic of China. 3. Department of Stomatology, Jinling Hospital, Jinling Medical School of Nanjing University, Medical School of Southern Medical University, Clinic School of Nanjing Medical University, Clinic School of the Fourth Military Medical University, Nanjing, Jiangsu 210002, People's Republic of China. Electronic address: zangshengqi@sina.com. 4. Department of Stomatology, Jinling Hospital, Jinling Medical School of Nanjing University, Medical School of Southern Medical University, Clinic School of Nanjing Medical University, Clinic School of the Fourth Military Medical University, Nanjing, Jiangsu 210002, People's Republic of China; State Key laboratory of Military Stomatology, Department of Prosthodontics, School of Stomatology, The Fourth Military Medical University, Xian, Shaanxi 710023, People's Republic of China. Electronic address: ljin@nju.edu.cn.
Abstract
AIMS: in vitro effects of bone morphogenetic protein 7 (BMP-7) on proliferation and differentiation of dental pulp stem cells (DPSCs) have not been investigated, nor has an appropriate dose been established. MAIN METHODS: Human DPSCs obtained from healthy volunteers were cultured with BMP-7 at 25, 50, and 100 ng/ml. Cell viability was measured by Cell Counting Kit-8 assay. Expression profiles of selected odontogenic differentiation-related markers in DPSCs were evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunocytochemistry, and western blot analysis. Mineralization of DPSCs was evaluated by alizarin red staining. The Smad5 signaling pathway was examined by qRT-PCR and western blot analysis. KEY FINDINGS: Diminished cell viability was found in DPSCs induced with 25, 50, and 100 ng/ml of BMP-7 for 7 days, showing a dose-response effect (P-trend = 0.03). DSPP, OCN, DMP-1, and RUNX2 were upregulated by BMP-7 induction after 7 and 14 days, especially at 50 and 100 ng/ml (P < 0.05). Immunocytochemical staining revealed strong expression of DSPP, DMP-1 and ALP in DPSCs induced by BMP-7, whereas null or weak expression in untreated cells. Western blot analysis confirmed over-expression of DSPP in cells induced by BMP-7. Alizarin red staining confirmed formation of mineralized nodules 4 weeks after BMP-7 induction. BMP-7 treated cells showed dose-dependently increased expression of BMPR1A, Smad5, and p-Smad5. SIGNIFICANCE: Our data indicated that BMP-7 at 50 ng/ml and 100 ng/ml was capable to induce DPSCs toward odontogenic differentiation through the Smad5 signaling pathway and not dramatically halt cell proliferation in vitro.
AIMS: in vitro effects of bone morphogenetic protein 7 (BMP-7) on proliferation and differentiation of dental pulp stem cells (DPSCs) have not been investigated, nor has an appropriate dose been established. MAIN METHODS:Human DPSCs obtained from healthy volunteers were cultured with BMP-7 at 25, 50, and 100 ng/ml. Cell viability was measured by Cell Counting Kit-8 assay. Expression profiles of selected odontogenic differentiation-related markers in DPSCs were evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunocytochemistry, and western blot analysis. Mineralization of DPSCs was evaluated by alizarin red staining. The Smad5 signaling pathway was examined by qRT-PCR and western blot analysis. KEY FINDINGS: Diminished cell viability was found in DPSCs induced with 25, 50, and 100 ng/ml of BMP-7 for 7 days, showing a dose-response effect (P-trend = 0.03). DSPP, OCN, DMP-1, and RUNX2 were upregulated by BMP-7 induction after 7 and 14 days, especially at 50 and 100 ng/ml (P < 0.05). Immunocytochemical staining revealed strong expression of DSPP, DMP-1 and ALP in DPSCs induced by BMP-7, whereas null or weak expression in untreated cells. Western blot analysis confirmed over-expression of DSPP in cells induced by BMP-7. Alizarin red staining confirmed formation of mineralized nodules 4 weeks after BMP-7 induction. BMP-7 treated cells showed dose-dependently increased expression of BMPR1A, Smad5, and p-Smad5. SIGNIFICANCE: Our data indicated that BMP-7 at 50 ng/ml and 100 ng/ml was capable to induce DPSCs toward odontogenic differentiation through the Smad5 signaling pathway and not dramatically halt cell proliferation in vitro.
Authors: Sarah Hani Shoushrah; Janis Lisa Transfeld; Christian Horst Tonk; Dominik Büchner; Steffen Witzleben; Martin A Sieber; Margit Schulze; Edda Tobiasch Journal: Int J Mol Sci Date: 2021-06-15 Impact factor: 5.923
Authors: Sara Martin-Iglesias; Lara Milian; María Sancho-Tello; Rubén Salvador-Clavell; José Javier Martín de Llano; Carmen Carda; Manuel Mata Journal: Stem Cells Int Date: 2022-03-04 Impact factor: 5.443