Different concentrations of C5a affect human dental pulp mesenchymal stem cells differentiation

Jie Liu,Mengtong Yuan,Xiaohan Tan,Junlong Hu,Li Gao,Weiping Hu,Xiaoling Wei,Mingyue Liu,Xin Shi,Ning Li,Xiaocui Kang

doi:10.1186/s12903-021-01833-4

Abstract

BackgroundDuring the process of deep decay, when decay approaches the pulp, an immune response is triggered inside the pulp, which activates the complement cascade. The effect of complement component 5a (C5a) on the differentiation of dental pulp mesenchymal stem cells (DPSCs) is related to dentin reparation. The aim of the present study was to stimulate DPSCs with different concentrations of C5a and evaluate the differentiation of odontoblasts using dentin sialoprotein (DSP).MethodsDPSCs were divided into the following six groups: (i) Control; (ii) DPSCs treated with 50 ng/ml C5a; (iii) DPSCs treated with 100 ng/ml C5a; (iv) DPSCs treated with 200 ng/ml C5a; (v) DPSCs treated with 300 ng/ml C5a; and (vi) DPSCs treated with 400 ng/ml C5a. Flow cytometry and multilineage differentiation potential were used to identify DPSCs. Mineralization induction, Real-time PCR and Western blot were conducted to evaluate the differentiation of odontoblast in the 6 groups.ResultDPSCs can express mesenchymal stem cell markers, including CD105, CD90, CD73 and, a less common marker, mesenchymal stromal cell antigen-1. In addition, DPSCs can differentiate into adipocytes, neurocytes, chondrocytes and odontoblasts. All six groups formed mineralized nodules after 28 days of culture. Reverse transcription-quantitative PCR and western blotting indicated that the high concentration C5a groups expressed higher DSP levels and promoted DPSC differentiation, whereas the low concentration C5a groups displayed an inhibitory effect.ConclusionIn this study, the increasing concentration of C5a, which accompanies the immune process in the dental pulp, has demonstrated an enhancing effect on odontoblast differentiation at higher C5a concentrations in vitro.

Highlights

During the process of deep decay, when decay approaches the pulp, an immune response is triggered inside the pulp, which activates the complement cascade
Identification of dental pulp mesenchymal stem cells (DPSCs) by CD34, CD45, CD73, CD90, CD105 and MSCA‐1 using flow cytometry Flow cytometry analysis indicated that CD34 (Fig. 1A), CD45 (Fig. 1B), CD73 (Fig. 1C), CD90 (Fig. 1D), CD105 (Fig. 1E) and Mesenchymal stem cell antigen (MSCA-1) (Fig. 1F) were expressed by 7.7, 2.9, 97.5, 91.5, 36.7 and 5.6% of DPSCs, respectively
The role of component 5a (C5a) following the migration of DPSCs, in particular, whether C5a continues to display a role during the second stage of regeneration has not been previously reported; the present study cultured DPSCs with different concentrations of C5a to determine the effects on DPSC differentiation

Summary

Introduction

During the process of deep decay, when decay approaches the pulp, an immune response is triggered inside the pulp, which activates the complement cascade. The effect of complement component 5a (C5a) on the differentiation of dental pulp mesenchymal stem cells (DPSCs) is related to dentin reparation. When deep decay approaches the pulp, bacterial metabolites stimulate the pulp via dentinal tubules, causing irritation of the pulp, even when the pulp is not exposed. During the early stages of stimulation, immune cells in the pulp that reach the damaged site release various inflammatory factors to attract more immune cells, resulting in cascade. The pulp is located in a relatively closed cavity, which makes the cycle of metabolism weaker; dead cells enhance the amplification of the inflammatory signal, and eventually, pulpitis occurs. A treatment strategy to control the pulpal immune response, induce repair and avoid the occurrence of pulpitis has not been identified

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