Effect of HUVECs on proliferation and stemness of hDPSCs silencing integrin α6

Objective To investigate the effect of human umbilical vein endothelial cells (HUVECs) on the proliferation and stemness of human dental pulp stem cells (hDPSCs) silencing with integrin α6 (ITGA6). Methods ITGA6 silencing lentivirus was used to interfere the ITGA6 expression of hDPSCs, and its silen...

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Main Author: AN Qi, ZHANG Weiwei, HE Lina, LI Yanping, PAN Shuang, NIU Yumei
Format: Article
Language:zho
Published: Editorial Office of Stomatology 2025-04-01
Series:Kouqiang yixue
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Online Access:https://www.stomatology.cn/fileup/1003-9872/PDF/1745561318749-630803018.pdf
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author AN Qi, ZHANG Weiwei, HE Lina, LI Yanping, PAN Shuang, NIU Yumei
author_facet AN Qi, ZHANG Weiwei, HE Lina, LI Yanping, PAN Shuang, NIU Yumei
author_sort AN Qi, ZHANG Weiwei, HE Lina, LI Yanping, PAN Shuang, NIU Yumei
collection DOAJ
description Objective To investigate the effect of human umbilical vein endothelial cells (HUVECs) on the proliferation and stemness of human dental pulp stem cells (hDPSCs) silencing with integrin α6 (ITGA6). Methods ITGA6 silencing lentivirus was used to interfere the ITGA6 expression of hDPSCs, and its silencing efficiency was detected. Then HUVECs were added to the chambers to co-culture, and the experiments were divided into four groups (sh-NC, sh-ITGA6, sh-NC+HUVECs and sh-ITGA6+HUVECs). hDPSCs in the sh-NC and sh-ITGA6 groups were transfected with sh-NC and sh-ITGA6 respectively. hDPSCs transfected with sh-NC and sh-ITGA6 were co-cultured with HUVECs in the sh-NC+HUVECs group and sh-ITGA6+HUVECs group respectively. The proliferation capacity of hDPSCs of each group was examined by CCK-8 and EdU on day 7. Immunofluorescence detected the expression of Stro-1, and Real-time PCR was used to detect the expression of Oct4 and Nanog. Results ①Fluorescence microscopy showed that the transfection efficiency was about 80%. Real-time PCR and Western blot results showed that sh-ITGA6 lentivirus effectively interfered with ITGA6 expression in hDPSCs. ②CCK-8 results showed that on day 5 of co-culture, the proliferation ability of the sh-ITGA6+HUVECs group was superior to that of the sh-ITGA6 group (P<0.05); on day 7, the proliferation ability of the sh-NC+HUVECs and sh-ITGA6+HUVECs group was superior to that of the sh-NC and sh-ITGA6 group (P<0.05). EdU results showed that the DNA synthesis ability of hDPSCs in the co-culture group was significantly stronger than that in the single-culture group (P<0.05). ③Immunofluorescence staining revealed that the expression of Stro-1 in the co-culture group was significantly stronger than that in the single-culture group. ④Real-time PCR results showed that the expression of Oct4 in the co-culture group was higher than that in the single-culture group (P<0.05); the expression of Nanog in hDPSCs with sh-ITGA6 was elevated by the addition of HUVECs co-culture (P<0.05). Conclusion HUVECs significantly enhance the proliferation and stemness of hDPSCs silencing integrin α6.
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spelling doaj-art-85086cd33e2f4df2aea4fc02fa8513e12025-08-20T03:54:01ZzhoEditorial Office of StomatologyKouqiang yixue1003-98722025-04-0145424825310.13591/j.cnki.kqyx.2025.04.002Effect of HUVECs on proliferation and stemness of hDPSCs silencing integrin α6AN Qi, ZHANG Weiwei, HE Lina, LI Yanping, PAN Shuang, NIU Yumei0Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin Medical University, School of Stomatology, Harbin 150001, ChinaObjective To investigate the effect of human umbilical vein endothelial cells (HUVECs) on the proliferation and stemness of human dental pulp stem cells (hDPSCs) silencing with integrin α6 (ITGA6). Methods ITGA6 silencing lentivirus was used to interfere the ITGA6 expression of hDPSCs, and its silencing efficiency was detected. Then HUVECs were added to the chambers to co-culture, and the experiments were divided into four groups (sh-NC, sh-ITGA6, sh-NC+HUVECs and sh-ITGA6+HUVECs). hDPSCs in the sh-NC and sh-ITGA6 groups were transfected with sh-NC and sh-ITGA6 respectively. hDPSCs transfected with sh-NC and sh-ITGA6 were co-cultured with HUVECs in the sh-NC+HUVECs group and sh-ITGA6+HUVECs group respectively. The proliferation capacity of hDPSCs of each group was examined by CCK-8 and EdU on day 7. Immunofluorescence detected the expression of Stro-1, and Real-time PCR was used to detect the expression of Oct4 and Nanog. Results ①Fluorescence microscopy showed that the transfection efficiency was about 80%. Real-time PCR and Western blot results showed that sh-ITGA6 lentivirus effectively interfered with ITGA6 expression in hDPSCs. ②CCK-8 results showed that on day 5 of co-culture, the proliferation ability of the sh-ITGA6+HUVECs group was superior to that of the sh-ITGA6 group (P<0.05); on day 7, the proliferation ability of the sh-NC+HUVECs and sh-ITGA6+HUVECs group was superior to that of the sh-NC and sh-ITGA6 group (P<0.05). EdU results showed that the DNA synthesis ability of hDPSCs in the co-culture group was significantly stronger than that in the single-culture group (P<0.05). ③Immunofluorescence staining revealed that the expression of Stro-1 in the co-culture group was significantly stronger than that in the single-culture group. ④Real-time PCR results showed that the expression of Oct4 in the co-culture group was higher than that in the single-culture group (P<0.05); the expression of Nanog in hDPSCs with sh-ITGA6 was elevated by the addition of HUVECs co-culture (P<0.05). Conclusion HUVECs significantly enhance the proliferation and stemness of hDPSCs silencing integrin α6.https://www.stomatology.cn/fileup/1003-9872/PDF/1745561318749-630803018.pdf|integrin α6|human dental pulp stem cells|umbilical vein endothelial cells|proliferation|stemness
spellingShingle AN Qi, ZHANG Weiwei, HE Lina, LI Yanping, PAN Shuang, NIU Yumei
Effect of HUVECs on proliferation and stemness of hDPSCs silencing integrin α6
Kouqiang yixue
|integrin α6|human dental pulp stem cells|umbilical vein endothelial cells|proliferation|stemness
title Effect of HUVECs on proliferation and stemness of hDPSCs silencing integrin α6
title_full Effect of HUVECs on proliferation and stemness of hDPSCs silencing integrin α6
title_fullStr Effect of HUVECs on proliferation and stemness of hDPSCs silencing integrin α6
title_full_unstemmed Effect of HUVECs on proliferation and stemness of hDPSCs silencing integrin α6
title_short Effect of HUVECs on proliferation and stemness of hDPSCs silencing integrin α6
title_sort effect of huvecs on proliferation and stemness of hdpscs silencing integrin α6
topic |integrin α6|human dental pulp stem cells|umbilical vein endothelial cells|proliferation|stemness
url https://www.stomatology.cn/fileup/1003-9872/PDF/1745561318749-630803018.pdf
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