Journal of Prevention and Treatment for Stomatological Diseases ›› 2018, Vol. 26 ›› Issue (5): 297-303.doi: 10.12016/j.issn.2096-1456.2018.05.004

• Basic Study • Previous Articles     Next Articles

Effects of inflammatory microenvironment mediated by macrophage on the proliferation and osteogenic differentiation of periodontal ligament cells

Xiumei ZHENG1(), Wenxia HUANG2()   

  1. 1. Department of Implantology, Stomatologic Hospital of Xiamen Medical college, Xiamen 361001, China
    2. Department of Periodontology, Stomatologic Hospital of Xiamen Medical College, Xiamen 361001, China
  • Received:2017-09-03 Revised:2017-10-01 Online:2018-05-20 Published:2018-08-30

Abstract:

Objective The present study investigated the effects of the inflammatory microenvironment mediated by macrophages on the proliferation and osteogenic differentiation of periodontal ligament cells (PDLCs). Methods Conditioned medium containing inflammatory factors was collected following macrophage activation with 1 μg/mL lipopolysaccharide (LPS). PDLCs were isolated from healthy teeth and cultured in conditioned medium (LPS-CM group) or normal medium (control group), and the proliferation of PDLCs was detected using the MTT assay. The cells were cocultured with an osteogenic inducer for 3 d and 7 d, and the alkaline phosphatase (ALP) activity of PDLCs was detected using an ALP kit. The mRNA expression levels of runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and collagen I (COL-I) were detected using real-time PCR, and the protein levels of RUNX2, OCN, and COL-I were detected using Western blotting. Mineralization nodules were observed using Alizarin red staining after osteoinduction for 14 d. Results The OD value of PDLCs in the LPS-CM group was lower than that in the control group (P < 0.05). The mRNA levels of RUNX2, OCN, and COL-I in the LPS-CM group were lower than those in the control group (P < 0.05). In addition to the OCN 3 d group (t = 2.75, P = 0.056), the protein expression of RUNX2, OCN, and COL-I in the LPS-CM group was lower than that in the control group (P < 0.05). However, the ALP activity of the LPS-CM group was higher than that of the control group, which was 1.58-fold greater (t = 5.91, P = 0.030) at 3 d and 1.29-fold greater (t = 6.01, P = 0.046) at 7 d. The number of calcified nodules in the LPS-CM group was significantly less than that in the control group (t = 8.63, P = 0.048). Conclusion The inflammatory microenvironment mediated by macrophages may inhibit the proliferation and osteogenic differentiation of PDLCs.

Key words: Macrophage, Lipopolysaccharide, Periodontal ligament cells, Inflammatory microenvironment, Osteogenic differentiation, Alkaline phosphatase, Osteocalcin, Collagen I

CLC Number: 

  • R781.4

Figure 1

Morphology of macrophage under inverted phase microscope × 100"

Figure 2

Gene expression of macrophage-related cytokines"

Figure 3

Growth curves of PDLCs of two groups"

Figure 4

ALP activity after osteogenic induction for 3 d and 7 d"

Figure 5

mRNA expression of Runx2, OCN and COL-1"

Figure 6

Protein expression of Runx2, OCN and COL-1 after osteogenic induction for 3, and 7 d"

Figure 7

Alizarin red staining results of PDLCs after osteogenic induction for 14 d and semi-quantitative analysis of calcium mineralization"

[1] 刘娟, 赵红宇, 轩东英. 人牙周膜细胞群多向分化潜能的实验研究[J]. 华西口腔医学杂志, 2010, 28(2): 185-189.
[2] Chang ZL.Recent development of the mononuclear phagocyte system: in memory of Metchnikoff and Ehrlich on the 100th Anniversary of the 1908 Nobel Prize in Physiology or Medicine[J]. Biol Cell, 2009, 101(12): 709-721.
[3] Cekici A, Kantarci A, Hasturk H, et al.Inflammatory and immune pathways in the pathogenesis of periodontal disease[J]. Periodontol 2000, 2014, 64(1): 57-80.
[4] Mosser DM, Edwards JP.Exploring the full spectrum of macrophage activation[J]. Nat Rev Immunol, 2008, 8(12): 958-969.
[5] Prieur X, Mok CY, Velagapudi VR, et al.Differential lipid partitioning between adipocytes and tissue macrophages modulates macrophage lipotoxicity and M2/M1 polarization in obese mice[J]. Diabetes, 2011, 60(3): 797-809.
[6] Han SW, Chen ZT.Activation of macrophages by lipopolysaccharide for assessing the immunomodulatory property of biomaterials[J]. Termis, 2015, 5(1): 19-28.
[7] Lacey D, Sampey A, Mitchell R, et al.Control of fibroblast-like synoviocyte proliferation by macrophage migration inhibitory factor[J]. Arthritis Rheum, 2003, 48(1): 103-109.
[8] Pacios S, Andriankaja O, Kang J, et al.Bacterial infection increases periodontal bone loss in diabetic rats through enhanced apoptosis[J]. Am J Pathol, 2013, 183(6): 1928-1935.
[9] 李颖, 张文元, 杨亚冬, 等. 人牙周膜细胞在矿化液作用下的增殖、成骨分化与相关基因表达[J]. 口腔医学研究, 2014, 1(1): 48-51, 56.
[10] Luca D, Carbonare L, Innamorati G, et al.Transcription factor RUNX2 and its application to hone tissue engineering[J]. Stem Cell Rev, 2012, 8(3): 891-897.
[11] 杨俊, 陈兴兴, 方煌, 等. LPS对人牙周膜细胞Toll样受体4及下游炎症因子表达的影响[J]. 临床口腔医学杂志, 2014, 30(11): 689-691.
[12] Kato H, Taguchi Y, Tominaga K, et al.Porphyromonas gingivalis LPS inhibits osteoblastic differentiation and promotes pro-inflammatory cytokine production in human periodontal ligament stem cells[J]. Arch Oral Biol, 2014, 59(2): 167-175.
[13] 袁萍, 李淑慧, 赵璐, 等. 炎症微环境下人牙周膜干细胞的生物学特性[J]. 中国组织工程研究, 2016, 20(6): 898-905.
[14] Ding J, Ghali O, Lencel P, et al.TNF-alpha and IL-1beta inhibit RUNX2 and collagen expression but increase alkaline phosphatase activity and mineralization in human mesenchymal stem cells[J]. Life Sci, 2009, 84(15/16): 499-504.
[15] Huang RL, Yuan Y, Zou GM, et al.LPS-stimulated inflammatory environment inhibits BMP-2-induced osteoblastic differentiation through crosstalk between TLR4/MyD88/NF-κB and BMP/Smad signaling[J]. Stem Cells Dev, 2014, 23(3): 277-289.
[16] Liu N, Shi ST, Deng MJ, et al.High levels of beta-Catenin signaling reduce osteogenic differentiation of stem cells in inflammatory microenvironments through inhibition of the noncanonical Wnt pathway[J]. J Bone Miner Res, 2011, 26(9): 2082-2095.
[17] Kong X, Liu Y.GSK3beta is a checkpoint for TNF-alpha mediated impaired osteogenic differentiation of mesenchymal stem cells in inflammatory microenvironments[J]. Biochim Biophys Acta, 2013, 1830(11): 5119-5129.
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