Effect of <em>Porphyromonas gingivalis</em> lipopolysaccharide on calcification of human umbilical artery smooth muscle cells co-cultured with human periodontal ligament cells

Li,Jing; Deng,Jing; Shang,Shuxian; Liu,Guirong; Song,Wenbin; Sun,Pei; Jiang,Wenjing; Pan,Keqing

doi:10.3892/etm.2021.10087

Effect of Porphyromonas gingivalis lipopolysaccharide on calcification of human umbilical artery smooth muscle cells co-cultured with human periodontal ligament cells

Authors:
- Jing Li
- Jing Deng
- Shuxian Shang
- Guirong Liu
- Wenbin Song
- Pei Sun
- Wenjing Jiang
- Keqing Pan
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Affiliations: Department of Stomatology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China, Department of Stomatology, Qingdao Municipal Hospital, Qingdao, Shandong 266011, P.R. China
Published online on: April 20, 2021 https://doi.org/10.3892/etm.2021.10087
Article Number: 655
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Periodontitis is an independent risk factor for coronary heart disease. Porphyromonas gingivalis lipopolysaccharide (Pg‑LPS) was considered to be one of the main virulence factors. In addition, vascular smooth muscle cells transform into osteoblast‑like cells in an arterial calcification process under chronic inflammatory conditions. The present study aimed to determine the calcification induced by Pg‑LPS in human umbilical artery smooth muscle cells (HUASMCs) co‑cultured with human periodontal ligament cells (HPDLCs). An in vitro co‑culture system was established using Transwell inserts. HUASMC proliferation and alkaline phosphatase (ALP) activity were measured with a Cell Counting Kit‑8 and an ALP kit, respectively. Calcium nodule formation was detected using alizarin red S staining. The effects of Pg‑LPS on the mRNA expression of the calcification genes of ALP, core‑binding factor α1 (Runx2) and bone sialoprotein (BSP) were assessed using reverse transcription‑quantitative PCR. The results indicated that Pg‑LPS increased HUASMC proliferation and ALP activity. Furthermore, among all of the groups, calcium nodule formation was most extensive in co‑cultured cells in the mineralization‑inducing medium containing Pg‑LPS. In addition, the expression of specific osteogenic genes (Runx2, ALP and BSP) significantly increased in the presence of Pg‑LPS and mineralization‑inducing medium, which was further enhanced in co‑culture with HPDLCs. In conclusion, co‑culture with HPDLCs increased the effect of Pg‑LPS to stimulate the calcification of HUASMCs. It was suggested that besides the inflammation, periodontitis may promote the occurrence of vascular calcification. The study indicated that periodontal treatment of subgingival scaling to reduce and/or control Porphyromonas gingivalis may decrease the occurrence or severity of vascular calcification.

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