“红色复合体”致病菌裂解液对人牙周膜细胞成骨分化能力的影响

doi:10.13701/j.cnki.kqyxyj.2020.09.010

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (9): 839-843.DOI: 10.13701/j.cnki.kqyxyj.2020.09.010

“红色复合体”致病菌裂解液对人牙周膜细胞成骨分化能力的影响

周何洁¹, 李胜鸿², 彭培钊², 唐榕², 李艾莲², 曾锦^3*

1.四川省新津县人民医院口腔科四川成都 610000;
2.西南医科大学口颌面修复重建和再生实验室四川泸州 646000;
3.西南医科大学口颌面修复重建和再生实验室,西南医科大学附属口腔医院正畸科四川泸州 646000

收稿日期:2020-02-15 出版日期:2020-09-28 发布日期:2020-09-15
通讯作者: *曾锦,E-mail: zengjin@swmu.edu.cn
作者简介:周何洁(1984~),女,四川新津人,硕士,主治医师,主要从事口腔疾病临床及基础研究。
基金资助:
四川省泸州市科技局应用基础研究项目(编号:2019-JYJ-55);国家级大学生创新创业训练计划项目(编号:201816032053);西南医科大学重点项目(编号:2017-ZRZD-001)

Effects of Extracts from “Red Complex” Periodontal Pathogens on Osteogenic Differentiation Capacity of Human Periodontal Ligament Cells

ZHOU Hejie¹, LI Shenghong², PENG Peizhao², TANG Rong², LI Ailian², ZENG Jin^3*

1. Department of Stomatology, Xinjin County People's Hospital, Chengdu 610000 China;
2. Orofacial Reconstruction and Regeneration Laboratory, The Affiliated Stomatological Hospital of Southwestern Medical University, Luzhou 646000, China;
3. Orofacial Reconstruction and Regeneration Laboratory, Department of Periodontal Orthodontics, The Affiliated Stomatological Hospital, Southwestern Medical University, Luzhou 646000, China

Received:2020-02-15 Online:2020-09-28 Published:2020-09-15

摘要/Abstract

摘要： 目的: 探讨“红色复合体”牙周致病菌裂解液对人牙周膜细胞成骨分化能力的影响。方法: 收集因正畸需要而拔除的健康前磨牙,原代培养人牙周膜干细胞(hPDLCs);采用免疫荧光方法和成骨分化诱导实验对培养的原代hPDLCs进行鉴定;体外联合培养牙龈卟啉单胞菌、齿垢密螺旋体和福赛斯坦纳菌,建立“红色复合体”模型,扫描电镜对其进行鉴定;制备“红色复合体”裂解液,通过碱性磷酸酶试剂盒、实时定量PCR、茜素红染色,检测裂解液对hPDLCs碱性磷酸酶活性,成骨分化相关基因表达的影响;以及对hPDLCs成骨矿化能力的影响。结果: 在“红色复合体”裂解液的刺激下,hPDLCs碱性磷酸酶活性,成骨分化相关基因ALP、Runx2、OPN、COL1的表达以及成骨矿化水平显著降低。结论: “红色复合体”牙周致病菌能够抑制hPDLCs的成骨分化能力。

关键词: 牙周膜细胞, 牙周致病菌, 红色复合体, 成骨分化能力

Abstract: Objective: To evaluate the effects of extracts from “Red Complex” periodontal pathogens on osteogenic differentiation capacity of human periodontal ligament cells (hPDLCs). Methods: hPDLCs were isolated and primary cultured from first premolars extracted from orthodontic patients. The primary cultured cells were verified by immunofluorescence and osteogenic induction methods. The model of “Red Complex” was set by co-culturing Pg, Td, and Tf in vitro, and was verified by SEM. Soluble bacterial extracts obtained from “Red Complex” was used to investigate the effects of “Red Complex” periodontal pathogens on the ALP activities, osteogenic related genes expression, and mineralization of hPDLCs by performing ALP assay, RT-PCR, and Alizarin Red staining. Results: Under the stimulation of 100 μg/mL bacterial extracts, the ALP activities, expression of osteogenic related genes ALP, Runx2, OPN, and COL1, and mineralization of hPDLCs were significantly decreased. Conclusion: “Red Complex” periodontal pathogens inhibited the osteogenic differentiation capacity of hPDLCs.

Key words: periodontal ligament cells, periodontal pathogens, Red Complex, osteogenic differentiation

周何洁, 李胜鸿, 彭培钊, 唐榕, 李艾莲, 曾锦. “红色复合体”致病菌裂解液对人牙周膜细胞成骨分化能力的影响[J]. 口腔医学研究, 2020, 36(9): 839-843.

ZHOU Hejie, LI Shenghong, PENG Peizhao, TANG Rong, LI Ailian, ZENG Jin. Effects of Extracts from “Red Complex” Periodontal Pathogens on Osteogenic Differentiation Capacity of Human Periodontal Ligament Cells[J]. Journal of Oral Science Research, 2020, 36(9): 839-843.

参考文献

[1] Kinane DF, Stathopoulou PG, Papapanou PN. Periodontal diseases [J]. Nat Rev Dis Primers,2017, 3:17038.
[2] Socransky SS, Haffajee AD, Cugini MA, et al. Microbial complexes in subgingival plaque [J]. J Clin Periodontol,1998, 25(2):134-144.
[3] Suzuki N, Yoneda M, Hirofuji T. Mixed red-complex bacterial infection in periodontitis [J]. J Dent,2013,2:587279.
[4] Bosshardt DD, Stadlinger B, Hendrik T. Cell-to-cell communication periodontal regeneration [J]. Clin Oral Implants Res, 2015, 26(3):229-239.
[5] 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.
[6] Trubiani O, Pizzicannella J, Caputi S, et al. Periodontal ligament stem cells: current knowledge and future perspectives [J]. Stem Cells Dev, 2019, 28(15):995-1003.
[7] Tanaka MH, Rodrigues TO, Finoti LS. The effect of conventional mechanical periodontal treatment on red complex microorganisms and clinical parameters in Down syndrome periodontitis patients: a pilot study [J]. Eur J Clin Microbiol, 2014, 34(3):601-608.
[8] 杨芳,陈明月,胡英英,等.PPARγ在脂多糖刺激牙周膜细胞中调控NF-κB信号通路的作用研究[J].口腔医学研究,2017,33(7):698-702.
[9] 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.
[10] Yu B, Li Q, Zhou M. LPS induced upregulation of the TLR4 signaling pathway inhibits osteogenic differentiation of human periodontal ligament stem cells under inflammatory conditions [J]. Int J Mol Med, 2019, 43(6):2341-2351.
[11] Albiero ML, Amorim BR, Casati MZ, et al. Osteogenic potential of periodontal ligament stem cells are unaffected after exposure to lipopolysaccharides [J]. Braz Oral Res, 2017, 31:e17.
[12] 谭咏梅,侯晋,杨小军,等.侵入细胞内的牙龈卟啉单胞菌影响人牙周膜细胞的增殖及成骨分化[J].南方医科大学学报,2016,36(4):525-531.
[13] D'Alimonte I, Mastrangelo F, Giuliani P, et al. Osteogenic differentiation of mesenchymal stromal cells: A comparative analysis between human subcutaneous adipose tissue and dental pulp [J]. Stem Cells Dev, 2017, 26(11):843-855.
[14] Borsani E, Salgarello S, Mensi M, et al. Histochemical and immunohistochemical evaluation of gingival collagen and metalloproteinases in peri-implantitis [J]. Acta Histochem, 2005, 107(3):231-240.
[15] Leung KS, Fung KP, Sher AH, et al. Plasma bone-specific alkaline phosphatase as an indicator of osteoblastic activity [J]. J Bone Joint Surg Br, 1993, 75(2):288-292.

“红色复合体”致病菌裂解液对人牙周膜细胞成骨分化能力的影响

Effects of Extracts from “Red Complex” Periodontal Pathogens on Osteogenic Differentiation Capacity of Human Periodontal Ligament Cells

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	王素文, 赖思煜, 习利军, 王利宏. 黄芪甲苷调节人牙周膜细胞炎症反应及成骨分化的作用及机制研究[J]. 口腔医学研究, 2020, 36(2): 121-125.
[2]	黄奕智, 郭俊, 杨健. 富血小板纤维蛋白对牙周膜成纤维细胞成骨能力影响的实验研究[J]. 口腔医学研究, 2020, 36(1): 41-45.
[3]	梁又德, 韦伟, 薛伟伟, 周瑞平, 傅润英, 王贻宁. 周期性张应力对牙周膜细胞内LIF和LIFR的表达影响[J]. 口腔医学研究, 2019, 35(9): 863-867.
[4]	程海燕, 沈兰花, 张瑞, 孟玲娜, 刘迪, 刑北昱, 陶冠男. 雌孕激素对人牙周膜细胞成骨分化影响的研究[J]. 口腔医学研究, 2019, 35(7): 657-660.
[5]	郑广明, 张健, 张文怡. 低频磁刺激对高糖诱导人牙周膜细胞损伤的保护作用[J]. 口腔医学研究, 2019, 35(12): 1177-1181.
[6]	沈兰花, 张瑞, 孟玲娜. 雌激素受体对去势大鼠牙周膜干细胞成骨分化能力的影响[J]. 口腔医学研究, 2018, 34(4): 448-451.
[7]	伍云, 赵艳, 季耀庭, 夏海滨, 张壁, 王贻宁. C4orf7在牙周膜细胞成骨分化中的作用及机制[J]. 口腔医学研究, 2018, 34(2): 125-129.
[8]	范芹, 管晓燕, 李小娜, 刘建国. TP对脂多糖介导人牙周膜成纤维细胞细胞间黏附分子-1表达的影响[J]. 口腔医学研究, 2018, 34(10): 1089-1092.
[9]	蒋思琪, 尹凤英, 张璠, 王敏, 夏海斌. 初级纤毛对人牙周膜细胞成骨相关基因表达的影响[J]. 口腔医学研究, 2018, 34(1): 10-13.
[10]	程孟文，周毅. MiR-34a在牙周膜细胞成骨向分化中的作用[J]. 口腔医学研究, 2017, 33(9): 928-932.
[11]	赵媛, 王姝, 韶波, 高颖. PTEN对H₂O₂诱导的牙周膜细胞增殖凋亡的影响[J]. 口腔医学研究, 2017, 33(8): 889-892.
[12]	杨芳, 陈明月, 胡英英, 汪昌宁. PPARγ在脂多糖刺激牙周膜细胞中调控NF-κB信号通路的作用研究[J]. 口腔医学研究, 2017, 33(7): 698-702.
[13]	徐明, 杨建斌. 红芪多糖对脂多糖诱导的牙周膜细胞凋亡及对Wnt信号通路的影响[J]. 口腔医学研究, 2017, 33(6): 646-649.
[14]	王锦华，张芳，李霞，陈庆勇. NF-κB非经典途径IKKα调控的Maspin在人牙周膜细胞中的表达[J]. 口腔医学研究, 2017, 33(10): 1052-1055.
[15]	李唐萍,边可胤,张向宇. 柠檬精油对牙周致病菌抑制作用及其安全性初步研究[J]. 口腔医学研究, 2017, 33(1): 99-101.