Necrostatin-1 promotes ectopic periodontal tissue like structure regeneration in LPS-treated PDLSCs.

Bingbing Yan,Kewen Wei,Yan Liu,Cheng Hu,Taiqiang Dai,Xinyu Qiu,Hongmei Zhang,Dehua Li,Yongchun Gu,Motohiro Komaki

doi:10.1371/journal.pone.0207760

Abstract

Necroptosis is a programmed necrosis, regulated by receptor interacting protein kinase 1(RIP1) and receptor interacting protein kinase 3(RIP3), and could be inhibited by necrostatin-1(Nec-1) specifically. This study aims to evaluate the effect of Nec-1 on LPS-treated periodontal ligament stem cells (PDLSCs). In the research, three groups were established: normal cultured PDLSCs, Porphyromonas gingivalis (Pg)-LPS stimulated PDLSCs and Pg-LPS+Nec-1 treated PDLSCs. The expression of RIP1 and RIP3 and osteogenic differentiation of PDLSCs in three groups were analyzed. Then, we constructed cell aggregates (CA) using PDLSCs, then PDLSCs-CA were combined with Bio-Oss in three groups were transplanted subcutaneously in nude mice to assess their potentials of periodontal tissue regeneration. The results showed that RIP1 and RIP3 were fully expressed in Pg-LPS stimulated PDLSCs and the level increased significantly. Nec-1 inhibited RIP1-RIP3 interaction, and further inhibited necroptosis of PDLSCs in inflammatory state. Moreover, Nec-1 pretreatment ameliorates the osteogenic differentiation of LPS-treated PDLSCs and can effectively promote the cementum like structure ectopic regenerative ability of PDLSCs in nude mice. These findings show RIP1/RIP3-mediated necroptosis is an important mechanism of cell death in PDLSCs. Nec-1 has a protective effect in reducing cell death and promotes ectopic periodontal tissue like structure regeneration by inhibiting necroptosis. Nec-1 is a hopeful therapeutic agent which protects cells from necroptosis and ameliorates functional outcome.

Highlights

Periodontitis is one of the most common oral diseases in adults, and the leading cause of adult tooth loss [1]
To test whether Nec-1 pretreatment had any effect on receptor interacting protein kinase 1 (RIP1)-RIP3 interaction in periodontal ligament stem cells (PDLSCs), we investigated the recruitment of RIP3 to RIP1 by immunoprecipitation
Our result showed that the interaction between RIP1 and RIP3 was greatly enforced in the Porphyromonas gingivalis (Pg)-LPS group

Summary

Introduction

Periodontitis is one of the most common oral diseases in adults, and the leading cause of adult tooth loss [1]. Its pathogenesis is due to the bacteria or their products that gather on the surface of tooth roots or periodontal tissue. It invades periodontal tissue, leading to chronic inflammation, absorption of alveolar bone and resulting in tooth loss [3]. PDLSCs play an important role in periodontal regeneration. They can differentiate into periodontal membrane cells and cementum cells and form the tissue of cementum /periodontal structure either in vitro, or in vivo [6, 7]. In this study we simulated periodontal inflammatory conditions by treatment of PDLSCs with Pg-LPS [12]

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Conclusion