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甲状旁腺激素分泌与受体表达决定牙髓干细胞成骨分化的年龄相关程度。

Parathyroid Hormone Secretion and Receptor Expression Determine the Age-Related Degree of Osteogenic Differentiation in Dental Pulp Stem Cells.

作者信息

Bhandi Shilpa, Alkahtani Ahmed, Reda Rodolfo, Mashyakhy Mohammed, Boreak Nezar, Maganur Prabhadevi C, Vishwanathaiah Satish, Mehta Deepak, Vyas Nishant, Patil Vikrant, Raj A Thirumal, Testarelli Luca, Patil Shankargouda

机构信息

Department of Restorative Dental Sciences, Division of Operative Dentistry, College of Dentistry, Jazan University, Jazan 45142, Saudi Arabia.

Department of Restorative Dental Sciences, College of Dentistry, King Saud University, Riyadh 11451, Saudi Arabia.

出版信息

J Pers Med. 2021 Apr 27;11(5):349. doi: 10.3390/jpm11050349.

Abstract

OBJECTIVE

To demonstrate the levels of parathyroid hormone secretion and genetic expressions of parathyroid hormone (PTH) and PTH1 receptor (PTH1R) genes in the dental pulp stem cells (DPSCs) from different age groups before and after induction of osteogenic differentiation. In addition, we also wanted to check their correlation with the degree of osteogenic differentiation.

METHODS

Human primary DPSCs from three age groups (milk tooth (SHEDs), 7-12 years old; young DPSCs (yDPSCs), 20-40 years old; old DPSCs (oDPSCs), 60+ years old) were characterized for mesenchymal stem cell (MSC) markers. DPSCs were subjected to osteogenic differentiation and functional staining. Gene expression levels were analyzed by qRT-PCR. Surface receptor analysis was done by flow cytometry. Comparative protein levels were evaluated by ELISA.

RESULTS

All SHEDs, yDPSCs, and oDPSCs were found to be expressing mesenchymal stem cell markers. SHEDs showed more mineralization than yDPSCs and oDPSCs after osteogenic induction. SHEDs exhibited higher expression of PTH and PTH1R before and after osteogenic induction, and after osteogenic induction, SHEDs showed more expression for RUNX2, ALPL, and OCN. Higher levels of PTH were observed in SHEDs and yDPSCs, and the number of PTH1R positive cells was relatively lower in yDPSCs and oDPSCs than in SHEDs. After osteogenic induction, SHEDs were superior in the secretion of OPG, and the secretions of ALPL and PTH and the number of PTH1R positive cells were relatively low in the oDPSCs.

CONCLUSIONS

The therapeutic quality of dental pulp stem cells is largely based on their ability to retain their stemness characteristics. This study emphasizes the criterion of aging, which affects the secretion of PTH by these cells, which in turn attenuates their osteogenic potential.

摘要

目的

展示不同年龄组牙髓干细胞(DPSCs)在成骨分化诱导前后甲状旁腺激素的分泌水平以及甲状旁腺激素(PTH)和甲状旁腺激素1型受体(PTH1R)基因的表达情况。此外,我们还想检查它们与成骨分化程度的相关性。

方法

对来自三个年龄组的人原代DPSCs(乳牙牙髓干细胞(SHEDs),7 - 12岁;年轻牙髓干细胞(yDPSCs),20 - 40岁;老年牙髓干细胞(oDPSCs),60岁以上)进行间充质干细胞(MSC)标志物鉴定。对DPSCs进行成骨分化和功能染色。通过qRT-PCR分析基因表达水平。通过流式细胞术进行表面受体分析。通过ELISA评估相对蛋白水平。

结果

发现所有SHEDs、yDPSCs和oDPSCs均表达间充质干细胞标志物。成骨诱导后,SHEDs比yDPSCs和oDPSCs表现出更多的矿化。SHEDs在成骨诱导前后PTH和PTH1R的表达均较高,成骨诱导后,SHEDs中RUNX2、ALPL和OCN的表达更多。在SHEDs和yDPSCs中观察到较高水平的PTH,yDPSCs和oDPSCs中PTH1R阳性细胞的数量比SHEDs中的相对较低。成骨诱导后,SHEDs在骨保护素(OPG)分泌方面更具优势,oDPSCs中碱性磷酸酶(ALPL)和PTH的分泌以及PTH1R阳性细胞的数量相对较低。

结论

牙髓干细胞的治疗质量很大程度上基于它们保持干性特征 的能力。本研究强调了衰老这一标准,其影响这些细胞中PTH的分泌,进而削弱它们的成骨潜能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba1d/8144966/7ea9b63b0493/jpm-11-00349-g001.jpg

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