Abstract:

Objective: To investigate the effects and molecular mechanism of static mechanical strain(SMS) on proliferation and osteogenic differentiation of human inflammatory periodontal ligament stem cells(PDLSCs). Methods: Healthy PDLSCs and inflammatory PDLSCs were treated with 10%, 12% and 14% SMS for 12 hours. Inflammatory PDLSCs were transinfected with negative control (NC) shRNA lentivirus, linc00638 shRNA lentivirus, miR-NC lentivirus or miR-29a inhibitor lentivirus for 72 hours, and then treated with 14% SMS for 12 hours. The optical density representing expression levels of long non coding RNA linc00638, miR-29a and proliferation activity were detected at 490 nm(A₄₉₀ level) by microplate reader. The alizarin red staining was used to detect the osteogenic differentiation at 14 days and the optical density at 405 nm (A₄₀₅ level) represented the level of osteogenic differentiation. Results: After 10%, 12% and 14% SMS intervention, compared with the baseline, the levels of A₄₉₀ and A₄₀₅ in healthy PDLSCs significantly increased(both P<0.05), and the expression levels of linc00638 and miR-29a did not change significantly(P>0.05); the levels of A₄₉₀, A₄₀₅ and the expression level of miR-29a in inflammatory PDLSCs significantly decreased, while the expression level of linc00638 significantly increased (all P<0.05). When 14% SMS intervention was given after knocking down linc00638, the levels of A₄₉₀, A₄₀₅ and the expression level of miR-29a in inflammatory PDLSCs significantly increased (all P<0.05). miR-29a significantly reduced the fluorescence activity of linc00638 demonstrated wild-type dual luciferase reporter in inflammatory PDLSCs(P<0.05). When 14% SMS intervention was given after knockdown of linc00638 and inhibition of miR-29a, the levels of A₄₉₀ and A₄₀₅ in inflammatory PDLSCs significantly decreased (both P<0.05). Conclusion: 14% SMS inhibits the proliferation and osteogenic differentiation of inflammatory PDLSCs by regulating the linc00638/miR-29a axis.

Key words: periodontal ligament stem cells, static mechanical strain, long non coding RNA, microRNA, proliferation, osteogenic differentiation

摘要：

目的： 研究静态牵张力（static mechanical strain，SMS）对人炎症牙周膜干细胞（periodontal ligament stem cells，PDLSCs）增殖及成骨分化的影响及分子机制。方法： 给予健康PDLSCs及炎症PDLSCs 10%、12%、14% SMS干预12 h，炎症PDLSCs感染阴性对照（NC）shRNA慢病毒、长链非编码RNA（long non coding RNA，lncRNA）linc00638 shRNA慢病毒、微小RNA（microRNA，miR）-NC慢病毒、miR-29a抑制物慢病毒72 h后给予14% SMS干预12 h。酶标仪在490 nm处检测吸光度值（A₄₉₀），代表linc00638、miR-29a的表达水平及增殖活力。成骨诱导分化14 d后进行茜素红染色，并在405 nm处检测吸光度值（A₄₀₅），代表PDLSCs成骨分化水平。结果： 在10%、12%、14% SMS的干预后，与干预前相比，健康PDLSCs的A₄₉₀、A₄₀₅明显增加（P<0.05），linc00638、miR-29a的表达水平无明显变化（P>0.05）；炎症PDLSCs的A₄₉₀、A₄₀₅及miR-29a的表达水平明显降低，linc00638的表达水平明显增加（P<0.05）。敲低linc00638后给予14%SMS干预，炎症PDLSCs A₄₉₀、A₄₀₅及miR-29a的表达水平明显增加（P<0.05）；miR-29a显著降低了炎症PDLSCs中linc00638野生型双荧光素酶报告基因的荧光活力（P<0.05）；敲低linc00638并抑制miR-29a后给予14%SMS干预，炎症PDLSCs的A₄₉₀、A₄₀₅明显降低（P<0.05）。结论： 14%SMS通过调控linc00638/miR-29a轴从而抑制炎症PDLSCs增殖及成骨分化。

关键词: 牙周膜干细胞, 静态牵张力, 长链非编码RNA, 微小RNA, 增殖, 成骨分化

CLC Number: 

• R782