Journal of Shandong University (Health Sciences) ›› 2021, Vol. 59 ›› Issue (3): 35-40.doi: 10.6040/j.issn.1671-7554.0.2021.0176

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Periodontal ligament stem cells regulate the functions of macrophages in vitro

ZHANG Ludan1, DING Xiaoling2, CUI Shuyue1, CHENG Chen1, WEI Fulan3, DING Gang1   

  1. 1. School of Stomatology, Weifang Medical University, Weifang 261053, Shandong, China;
    2. Clinical Competency Training Center, Weifang Medical University, Weifang 261053, Shandong, China;
    3. School of Stomatology, Shandong University, Jinan 250012, Shandong, China
  • Published:2021-04-06

Abstract: Objective To explore the effects of periodontal ligament stem cells(PDLSCs)on the phenotypes and functions of macrophages. Methods After PDLSCs were isolated and cultured, the expression profiles of STRO-1, CD146 and CD90, as well as the multipotent differentiation capabilities were detected. After macrophages were isolated from peripheral blood, they were cocultured with an equal amount PDLSCs in Transwell co-culture condition at 37 ℃ and 5% CO2, which were set as the experimental group. Macrophages cultured alone were set as the control group. After 3d co-culture, the expression profiles of CD14+CD206+ macrophages were examined by flow cytometry. After 24 h co-culture, macrophages were obtained, fluorescein isothiocyanate labeled dextran was added. Then, after 30 min incubation, the phagocytosis rate of macrophages was detected with flow cytometry. After 3 d co-culture, the supernatant was collected, and the concentrations of IL-10, IL-6 and TNF-α were determined with enzyme-linked immunosorbent assays. Results PDLSCs displayed fusiform fibroblast-like morphology, positive for the mesenchymal stem cells surface markers including STRO-1, CD146 and CD90, and could differentiate into bone cells and lipid cells. Compared with the control group, the experimental group had significantly increased expression of CD14+CD206+macrophages [(38.73±6.32)% vs(8.39±2.71)%, t=127.7, P=0.004 9), unchanged phagocytosis rate of macrophages [(36.7±5.1)% vs(38.6±4.3)%, t=3.904, P=0.159 6], elevated level of IL-10 [(382.5±18.2)pg/mL vs(198.5±11.4)pg/mL, t=76.36, P=0.000 3], but decreased levels of IL-6 [(453.1±70.42)pg/mL vs(936.7±49.9)pg/mL, t=53.12, P=0.011 5)and TNF-α [(64.9±11.3)pg/mL vs(131.7±19.3)pg/mL, t=51.48, P=0.000 6]. Conclusion PDLSCs are capable of converting macrophages into M2 phenotype without affecting the phagocytic functions. Meanwhile, they can stimulate the secretion of IL-10 but inhibit the secretion of IL-6 and TNF-α.

Key words: Periodontal ligament stem cells, Macrophages, Polarization, Transwell co-culture system

CLC Number: 

  • R781
[1] Kassebaum NJ, Bernabé E, Dahiya M, et al. Global burden of severe periodontitis in 1990-2010: a systematic review and meta-regression [J]. J Dent Res, 2014, 93(11): 1045-1053.
[2] Sanz M, Marco Del Castillo A, Jepsen S, et al. Periodontitis and cardiovascular diseases: Consensus report [J]. J Clin Periodontol, 2020, 47(3): 268-288.
[3] Baeza M, Morales A, Cisterna C, et al. Effect of periodontal treatment in patients with periodontitis and diabetes: systematic review and meta-analysis [J]. J Appl Oral Sci, 2020, 28: e20190248. doi:10.1590/1678-7757-2019-0248.
[4] Kocher T, König J, Borgnakke WS, et al. Periodontal complications of hyperglycemia/diabetes mellitus: Epidemiologic complexity and clinical challenge [J]. Periodontol 2000, 2018, 78(1): 59-97.
[5] Opacic J, Maldonado A, Ramseier CA, et al. Influence of periodontitis on pregnancy and childbirth [J]. Swiss Dent J, 2019, 129(7/8): 581-589.
[6] Manresa C, Sanz-Miralles EC, Twigg J, et al. Supportive periodontal therapy(SPT)for maintaining the dentition in adults treated for periodontitis [J]. Cochrane Database Syst Rev, 2018, 1: CD009376.
[7] Seo BM, Miura M, Gronthos S, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament [J]. Lancet, 2004, 364(9429): 149-155.
[8] Xu XY, Li X, Wang J, et al. Periodontal tissue regeneration using stem cells: strategies and translational considerations [J]. Stem Cells Transl Med, 2019, 8(4): 392-403.
[9] Liu Y, Zheng Y, Ding G, et al. Periodontal ligament stem cell-mediated treatment for periodontitis in miniature swine [J]. Stem Cells, 2008, 26(4): 1065-1073.
[10] Ding G, Liu Y, Wang W, et al. Allogeneic periodontal ligament stem cell therapy for periodontitis in swine [J]. Stem Cells, 2010, 28(10): 1829-1838.
[11] Tomokiyo A, Wada N, Maeda H. Periodontal ligament Stem cells: regenerative potency in periodontium [J]. Stem Cells Dev, 2019, 28(15):974-985.
[12] Gao B, Deng R, Chai Y, et al. Macrophage-lineage TRAP+cells recruit periosteum-derived cells for periosteal osteogenesis and regeneration [J]. J Clin Invest, 2019, 129(6): 2578-2594.
[13] Theret M, Mounier R, Rossi F. The origins and non-canonical functions of macrophages in development and regeneration [J]. Development, 2019, 146(9): dev156000.
[14] Yang ZP, Li Q, Wang X, et al. C-type lectin receptor LSECtin-mediated apoptotic cell clearance by macrophages directs intestinal repair in experimental colitis [J]. Proc Natl Acad Sci USA, 2018, 115(43): 11054-11059.
[15] Tang R, Wei F, Wei L, et al. Osteogenic differentiated periodontal ligament stem cells maintain their immunomodulatory capacity [J]. J Tissue Eng Regen Med, 2014, 8(3): 226-232.
[16] Liu D, Xu J, Liu O, et al. Mesenchymal stem cells derived from inflamed periodontal ligaments exhibit impaired immunomodulation [J]. J Clin Periodontol, 2012, 39(12): 1174-1182.
[17] Liu O, Xu J, Ding G, et al. Periodontal ligament stem cells regulate B lymphocyte function via programmed cell death protein 1 [J]. Stem Cells, 2013, 31(7):1371-1382.
[18] Murray PJ, Allen JE, Biswas SK, et al. Macrophage activation and polarization: nomenclature and experimental guidelines [J]. Immunity, 2014, 41(1): 14-20.
[19] Siamon Gordon, Fernando O Martinez. Alternative activation of macrophages: mechanism and functions [J]. Immunity, 2010, 32(5): 593-604.
[20] Shapouri-Moghaddam A, Mohammadian S, Vazini H, et al. Macrophage plasticity, polarization, and function in health and disease [J]. J Cell Physiol, 2018, 233(9): 6425-6440.
[21] Yunna C, Mengru H, Lei W, et al. Macrophage M1/M2 polarization [J]. Eur J Pharmacol, 2020, 877: 173090. doi: 10.1016/j.ejphar.2020.173090.
[22] Orecchioni M, Ghosheh Y, Pramod AB, et al. Macrophage polarization: different gene signatures in M1(LPS+)vs. classically and M2(LPS-)vs. alternatively activated macrophages [J]. Front Immunol, 2019, 10: 1084. doi:10.3389/fimmu.2019.01084.
[23] Kovach TK, Dighe AS, Lobo PI, et al. Interactions between MSCs and immune cells: implications for bone healing [J]. J Immunol Res, 2015, 2015: 752510. doi:10.1155/2015/752510.
[24] Pajarinen J, Lin T, Gibon E, et al. Mesenchymal stem cell-macrophage crosstalk and bone healing [J]. Biomaterials, 2019, 196: 80-89. doi:10.1016/j.biomaterials.2017.12.0.
[25] Nemeth K, Leelahavanichkul A, Yuen PS, et al. Bone marrow stromal cells attenuate sepsis via prostaglandin E(2)-dependent reprogramming of host macrophages to increase their interleukin-10 production [J]. Nat Med, 2009, 5(1): 42-49.
[26] Morand DN, Davideau JL, Clauss F, et al. Cytokines during periodontal wound healing: potential application for new therapeutic approach [J]. Oral Dis, 2017, 23(3): 300-311.
[27] Shi M, Wang C, Wang Y, et al. Deproteinized bovine bone matrix induces osteoblast differentiation via macrophage polarization [J]. J Biomed Mater Res A, 2018, 106(5): 1236-1246.
[28] Qiu X, Liu S, Zhang H, et al. Mesenchymal stem cells and extracellular matrix scaffold promote muscle regeneration by synergistically regulating macrophage polarization toward the M2 phenotype [J]. Stem Cell Res Ther, 2018, 9(1): 88.
[29] Gao X, Shen Z, Guan M, et al. Immunomodulatory role of stem cells from human exfoliated deciduous teeth on periodontal regeneration [J]. Tissue Eng Part A, 2018, 24(17/18): 1341-1353.
[30] 高弘斐, 张潜, 陈龙, 等. 间充质干细胞与巨噬细胞共培养体系的细胞因子表达模式研究[J]. 免疫学杂志, 2017, 33(11): 930-936. GAO Hongfei, ZHANG Qian, CHEN Long, et al. The expression pattern of cytokines in mesenchymal stem cells and macrophages co-cultured system[J]. Immunological Journal, 2017, 33(11): 930-936.
[31] 孙瑶, 吕海金, 易小猛, 等. 间充质干细胞通过诱导M2型巨噬细胞治疗急性肺损伤[J].中山大学学报(医学版), 2019, 40(3): 393-400. SUN Yao, LV Haijin, YI Xiaomeng, et al. Mesenchymal stem cells attenuate acute lung injury through inducing M2 macrophage polarization [J]. Journal of Sun Yatsen Univerty(Medical Sciences), 2019, 40(3): 393-400.
[32] Philipp D, Suhr L, Wahlers T,et al.Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization [J]. Stem Cell Res Ther, 2018, 9(1): 286.
[33] Wang Y, Han B, Wang Y, et al. Mesenchymal stem cell-secreted extracellular vesicles carrying TGF-β1 up-regulate miR-132 and promote mouse M2 macrophage polarization [J]. Cell Mol Med, 2020, 24(21): 12750-12764.
[34] Gong M, Zhuo X, Ma A. STAT6 Upregulation Promotes M2 Macrophage Polarization to Suppress Atherosclerosis [J]. Med Sci Monit Basic Res, 2017, 23: 240-249. doi: 10.12659/msmbr.904014.
[35] Liu F, Qiu H, Xue M, et al. MSC-secreted TGF-β regulates lipopolysaccharide-stimulated macrophage M2-like polarization via the Akt/FoxO1 pathway [J]. Stem Cell Res Ther, 2019, 10(1): 345.
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