表面纳米形貌对牙周膜干细胞衰老的作用研究

doi:10.7518/hxkq.2024.2023244

华西口腔医学杂志 ›› 2024, Vol. 42 ›› Issue (2): 172-180.doi: 10.7518/hxkq.2024.2023244

表面纳米形貌对牙周膜干细胞衰老的作用研究

孙艳萍(), 廖立()

口腔疾病防治全国重点实验室国家口腔医学中心国家口腔疾病临床医学研究中心教育部口腔转化医学工程研究中心四川大学华西口腔医院，成都 610041

收稿日期:2023-08-07 修回日期:2023-09-16 出版日期:2024-04-01 发布日期:2024-03-26
通讯作者: 廖立 E-mail:2021224030011@stu.scu.edu.cn;lliao@scu.edu.cn
作者简介:孙艳萍，硕士，E-mail：2021224030011@stu.scu.edu.cn
基金资助:
国家重点研发计划(2021YFA1100603);国家自然科学基金(82071092);四川省重点研发项目(2023YFS0056)

Effects of surface nanomorphology on the senescence of periodontal ligament stem cells

Sun Yanping(), Liao Li()

State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases & Engineering Research Center of Oral Translational Medicine, Ministry of Education & West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China

Received:2023-08-07 Revised:2023-09-16 Online:2024-04-01 Published:2024-03-26
Contact: Liao Li E-mail:2021224030011@stu.scu.edu.cn;lliao@scu.edu.cn
Supported by:
National Key R&D Program of China(2021YFA1100603);National Nature Science Foundation of China(82071092);Key R&D Projects of Sichuan Province(2023YFS0056)

摘要/Abstract

摘要：

目的探讨二氧化钛纳米管形貌对衰老牙周膜干细胞分化能力的影响。方法利用阳极氧化法，分别于20、70 V电压下制备出2种具有二氧化钛纳米管形貌的钛片（20V-NT、70V-NT），观察其表面形貌特征。在成骨诱导条件下培养年轻牙周膜干细胞，挑选具有促进成骨分化作用的表面形貌。用RO3306和Nutlin-3a诱导年轻牙周膜干细胞衰老，获得衰老的牙周膜干细胞。诱导衰老牙周膜干细胞成骨分化，观察表面形貌对衰老牙周膜干细胞成骨分化的影响。结果阳极氧化法可在钛片表面形成纳米管形貌，且纳米管直径随电压的增大而增大；不同直径纳米管形貌对年轻牙周膜干细胞成骨分化的影响存在较大差异，20V-NT表面纳米形貌促成骨分化效果更明显。与光滑钛片相比，20V-NT表面纳米形貌提高了衰老牙周膜干细胞碱性磷酸酶阳性数量，促进了钙沉积以及成骨标志性基因Runt相关转录因子2、骨桥蛋白、骨钙素的表达。结论特定的表面纳米形貌能增强衰老牙周膜干细胞的分化能力，为牙周再生和进一步提高种植体的性能提供了一种有效方法。

关键词: 表面形貌, 钛, 纳米管, 成骨, 衰老, 牙周膜干细胞

Abstract:

Objective The effect of TiO₂ nanotube morphology on the differentiation potency of senescent periodontal ligament stem cells was investigated. Methods Two types of titanium sheets with TiO₂nanotube morphology (20V-NT and 70V-NT) were prepared via anodic oxidation at 20 and 70 V separately, and their surface morphology was observed. Young periodontal ligament stem cells were cultivated in an osteogenic induction medium, and the most effective surface morphology in promoting osteogenic differentiation was selected. RO3306 and Nutlin-3a were used to induce the aging of young periodontal ligament stem cells, and senescent periodontal ligament stem cells were obtained. The osteogenic differentiation of senescent periodontal ligament stem cells was induced, and the effect of surface morphology on osteogenic differentiation was observed. Results Nanotube morphology was achieved on the surfaces of titanium sheets through anodic oxidation, and the diameters of the nanotubes increased with voltage. A significant difference in the effect of nanotube morphology was found among nanotubes with different diameters in the young periodontal ligament stem cells. The surface nanotube morphology of 20V-NT had a more significant effect that promoted osteogenic differentiation. Compared with a smooth titanium sheet, the surface nanotube morphology of 20V-NT increased the number of alkaline phosphatase-positive senescent periodontal ligament stem cells and promoted calcium deposition and the expression of osteogenic marker genes Runt-related transcription factor 2, osteopontin, and osteocalcin. Conclusion A special nanotube morphology enhances the differentiation ability of senescent periodontal ligament stem cells, provides an effective method for periodontal regeneration, and further improves the performance of implants.

Key words: surface morphology, titanium, nanotubes, osteogenesis, senescence, periodontal ligament stem cells

中图分类号:

R318.08

孙艳萍, 廖立. 表面纳米形貌对牙周膜干细胞衰老的作用研究[J]. 华西口腔医学杂志, 2024, 42(2): 172-180.

Sun Yanping, Liao Li. Effects of surface nanomorphology on the senescence of periodontal ligament stem cells[J]. West China Journal of Stomatology, 2024, 42(2): 172-180.

图/表 5

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基因	引物（5’-3’）
RUNX2	上游：TGGTTACTGTCATGGCGGGTA
	下游：TCTCAGATCGTTGAACCTTGCTA
OPN	上游：GAAGTTTCGCAGACCTGACAT
	下游：GTATGCACCATTCAACTCCTCG
OCN	上游：CACTCCTCGCCCTATTGGC
	下游：CCCTCCTGCTTGGACACAAAG
BMP2	上游：ACCCGCTGTCTTCTAGCGT
	下游：TTTCAGGCCGAACATGCTGAG
MKI67	上游：TCCTTTGGTGGGCACCTAAGACCTG
	下游：TGATGGTTGAGGTCGTTCCTTGATG
GAPDH	上游：CTTTGGTATCGTGGAAGGACTC
	下游：GTAGAGGCAGGGATGATGTTCT

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表面纳米形貌对牙周膜干细胞衰老的作用研究

Effects of surface nanomorphology on the senescence of periodontal ligament stem cells

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