Journal of Prevention and Treatment for Stomatological Diseases ›› 2022, Vol. 30 ›› Issue (5): 321-329.DOI: 10.12016/j.issn.2096-1456.2022.05.003

• Basic Study • Previous Articles     Next Articles

Effects of a red light-emitting diode on the proliferation and osteogenic differentiation of stem cells from the apical papilla

SU Yutong(), HOU Lan, JIANG Bing, ZHENG Genzi, LIU Yuan, WANG Yao()   

  1. The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, China
  • Received:2021-09-08 Revised:2021-12-10 Online:2022-05-20 Published:2022-02-17
  • Contact: WANG Yao
  • Supported by:
    Luzhou Municipal People’s Government-Southwest Medical University Science and Technology Strategic Cooperation Projects(2017LZXNYD-T03);Scientific Project of Southwest Medical University(2021ZKMS013)

LED红光对人根尖乳头干细胞增殖和成骨分化的影响

苏雨童(), 侯兰, 姜冰, 郑艮子, 刘源, 王瑶()   

  1. 西南医科大学附属口腔医院,四川 泸州(646000)
  • 通讯作者: 王瑶
  • 作者简介:苏雨童,硕士研究生,Email: syt247934557@qq.com
  • 基金资助:
    泸州市人民政府-西南医科大学科技战略合作项目(2017LZXNYD-T03);西南医科大学校级科研项目(2021ZKMS013)

Abstract:

Objective To explore the effects of red LEDs on the proliferation and osteogenic differentiation of human stem cells from apical papilla (hSCAPs). Methods hSCAPs were obtained by isolation, culture and flow cytometry in vitro and irradiated with 1, 3, 5, and 7 J/cm2 red LEDs. The proliferation of hSCAPs was detected using a CCK-8 assay. The osteogenic differentiation of hSCAPs was evaluated using alkaline phosphatase (ALP) staining, ALP activity assay and Alizarin red quantitative detection. The effect of 5 J/cm2 red LEDs on the expression levels of the ALP, Runx2, OCN, OPN and BSP genes and proteins was detected by RT-PCR and western blot, respectively. Results Red LEDs at 1, 3, 5, and 7 J/cm2 promoted the proliferation of hSCAPs (P < 0.05). The effects of red LEDs with different light energies on the proliferation of hSCAPs were different at different time points (P < 0.05). On the 7th and 14th days after irradiation, red LEDs promoted the osteogenic differentiation of hSCAPs, and the effect of 5 J/cm2 red LEDs was the most obvious under osteogenic induction culture conditions (P<0.05). Red LEDs (5 J/cm2) promoted the expression of the ALP, Runx2, OCN, OPN and BSP genes and proteins (P < 0.05). Conclusion Red LEDs promoted the proliferation and osteogenic differentiation of hSCAPs.

Key words: mesenchymal stem cells, human stem cells from apical papilla, photobiomodulation, red LED, cell proliferation, alkaline phosphatase, osteocalcin, osteopontin, osteogenic differentiation, tissue engineering, stem cell-based therapy

摘要:

目的 探讨LED红光对人根尖乳头干细胞(human stem cells from apical papilla,hSCAPs)增殖和成骨分化的影响。方法 hSCAPs通过体外分离培养和流式鉴定获得,分别用1、3、5、7 J/cm2 LED红光照射。CCK-8检测细胞增殖。碱性磷酸酶(alkaline phosphatase,ALP)染色及定量活性检测、茜素红定量检测分析成骨分化。RT-PCR实验和Western blot实验分别检测5 J/cm2 LED红光对hSCAPs中ALP、Runt相关转录因子2(Runt-related transcription factor 2,Runx2)、骨钙素(osteocalcin,OCN)、骨桥蛋白(osteopontin,OPN)和骨唾液酸蛋白(bone sialoprotein,BSP)基因及蛋白表达水平的影响。结果 1、3、5、7 J/cm2 LED红光照射促进hSCAPs的增殖(P<0.05);不同能量LED红光在不同的时间点照射对hSCAPs增殖的促进作用有差异(P<0.05);在成骨诱导培养下,光照后第7天和第14天,LED红光照射促进hSCAPs的成骨分化,且5 J/cm2 LED红光照射促进作用最明显(P<0.05);5 J/cm2 LED红光照射上调SCAPs中ALP、Runx2、OCN、OPN和BSP基因及蛋白的表达(P<0.05)。 结论 LED红光照射促进hSCAPs增殖和成骨分化。

关键词: 间充质干细胞, 人根尖乳头干细胞, 光生物调节, LED红光, 细胞增殖, 碱性磷酸酶, 骨钙素, 骨桥蛋白, 成骨分化, 组织工程, 干细胞治疗

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