牙龈卟啉单胞菌精氨酸特异性牙龈素基因疫苗预防比格犬种植体周围炎的实验研究

doi:10.7518/hxkq.2018.01.015

华西口腔医学杂志 ›› 2018, Vol. 36 ›› Issue (1): 76-81.doi: 10.7518/hxkq.2018.01.015

牙龈卟啉单胞菌精氨酸特异性牙龈素基因疫苗预防比格犬种植体周围炎的实验研究

李传花¹(), 王志峰², 朱丽娜², 范欣³, 蓝菁¹()

1.山东大学口腔医学院,山东省口腔组织再生重点实验室
2.山东大学口腔医院儿童牙病科,济南 250012
3.潍坊医学院附属医院口腔科,潍坊 261031

收稿日期:2017-05-25 修回日期:2017-08-02 出版日期:2018-02-07 发布日期:2018-02-01
作者简介:
李传花,住院医师,硕士,E-mail:lichuanhua1022@126.com
基金资助:
[基金项目] 国家自然科学基金（30801308）;山东省科技厅攻关项目（2010G0020237）

Experimental research on Arginine-gingipain A gene vaccine from Porphyromonas gingivalis that prevents peri-implantitis in Beagle dogs

Chuanhua Li¹(), Zhifeng Wang², Lina Zhu², Xin Fan³, Jing Lan¹()

1. School of Stomatology, Shandong University, Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan 250012, China
2. Dept. of Pediatrics Dentistry, School of Stomatology, Shandong University, Jinan 250012, China
3. Dept. of Stomatology, Weifang Medical School Affiliated Hospital, Weifang 261031, China

Received:2017-05-25 Revised:2017-08-02 Online:2018-02-07 Published:2018-02-01
Supported by:
Supported by: The National Natural Science Foundation of China (30801308);Foundation of Department of Science and Technology of Shandong Province (2010G0020237).

摘要/Abstract

摘要：

目的构建牙龈卟啉单胞菌精氨酸特异性牙龈素基因疫苗pVAX1-rgpA,并对成年犬进行免疫接种,观察该疫苗在防治种植体周围炎发生发展中的作用。方法构建真核表达质粒pVAX1-rgpA。拔除15只成年犬下颌双侧第二、三前磨牙,随机即刻植入种植体。3个月后,实验犬随机均分成A、B、C组,分别接种重组质粒pVAX1-rgpA、热失活牙龈卟啉单胞菌、空白质粒pVAX1,连续接种3次。接种开始前、接种结束2周后采用酶联免疫吸附试验检测血清IgG和唾液分泌型IgA（sIgA）含量。随机选取一侧采用丝线结扎法构建种植体周围炎,并检测种植体周探诊深度（PD）和探诊出血指数（BOP）。结扎6周后处死所有动物,制作50 μm厚的硬组织切片,亚甲基蓝染色后观察种植体周骨丧失程度。结果 A、B组动物免疫后,IgG、sIgA抗体较未免疫前明显增高（P<0.05）,同时较C组升高（P<0.05）,但A、B组间差异无统计学意义（P>0.05）。丝线结扎第4和6周时,C组结扎侧的PD明显高于A、B组（P<0.05）,A、B组间差异无明显统计学意义（P>0.05）。A组结扎侧骨丧失量明显小于其他两组（P<0.05）。硬组织切片可见,各组种植体周骨丧失区均有大量的炎症细胞和细菌存在,C组结扎侧最严重。结论精氨酸特异性牙龈素（rgpA）基因疫苗产生的IgG和sIgA,能有效减弱犬种植体周围炎的骨丧失量。

关键词: 基因疫苗, 牙种植体, 种植体周围炎, 精氨酸特异性牙龈素, 牙龈卟啉单胞菌

Abstract:

Objective This study aims to use Arginine-gingipain A gene vaccine (pVAX1-rgpA) to immunize adult Beagle dogs and to evaluate its effect during peri-implantitis progression and development. Methods Plasmid pVAX1-rgpA was constructed. The second and third bilateral mandible premolars of 15 adult Beagle dogs were extracted, and the implants were placed immediately. After 3 months, the animals were randomly divided into groups A, B, and C. Afterward, the animals were immunized thrice with plasmid pVAX1-rgpA, with heat-killed Porphyromonas gingivalis, or pVAX1, respectively. IgG in the serum and secretory IgA (sIgA) in saliva were quantitatively analyzed by enzyme-linked immunosorbent assay before and after 2 weeks of immunization. Peri-implantitis was induced with cotton ligatures fixed around the neck of implants. Probing depth (PD) and bleeding on probing were recorded. All animals were sacrificed after ligaturation for 6 weeks. Decalcified sections with thickness of 50 μm were prepared and dyed with methylene blue to observe the bone phenotype around implants. Results Levels of serum IgG and sIgA in saliva were higher in groups A and B after immunization than before the process (P<0.05) and higher than those in group C (P<0.05). However, no difference was observed between groups A and B (P>0.05). At 4 and 6 weeks after ligaturation, PD of the ligatured side in group C was higher than that in groups A and B (P<0.05). On the other hand, no difference was identified between groups A and B (P>0.05). Bone loss in group A was sig-nificantly lower than that of the other groups (P<0.05). Abundant inflammatory cells and bacteria were present in the bone loss area around the implants in the three groups, as identified through hard tissue section observation. However, group C presented the most number of inflammatory cells and bacteria in the bone loss area around the implants. Conclusion IgG and sIgA can be generated by immunity with rgpA DNA vaccine, which can significantly slow down bone loss during expe-rimental peri-implantitis in dogs.

Key words: gene vaccine, dental implant, peri-implantitis, Arginine-gingipains, Porphyromonas gingivalis

中图分类号:

R783

李传花, 王志峰, 朱丽娜, 范欣, 蓝菁. 牙龈卟啉单胞菌精氨酸特异性牙龈素基因疫苗预防比格犬种植体周围炎的实验研究[J]. 华西口腔医学杂志, 2018, 36(1): 76-81.

Chuanhua Li, Zhifeng Wang, Lina Zhu, Xin Fan, Jing Lan. Experimental research on Arginine-gingipain A gene vaccine from Porphyromonas gingivalis that prevents peri-implantitis in Beagle dogs[J]. West China Journal of Stomatology, 2018, 36(1): 76-81.

图/表 6

图 1

图 2

表 1

图 3

表 2

图 4

参考文献 15

[1]	Corbella S, Del Fabbro M, Taschieri S, et al.Clinical eva-luation of an implant maintenance protocol for the prevention of peri-implant diseases in patients treated with immediately loaded full-arch rehabilitations[J]. Int J Dent Hyg, 2011, 9
	(3):216-222.
[2]	Paramonov N, Rangarajan M, Hashim A, et al.Structural analysis of a novel anionic polysaccharide from Porphyro-monas gingivalis strain W50 related to Arg-gingipainglycans[J].Mol Microbiol, 2005, 58(3):847-863.
[3]	Yamatake K, Maeda M, Kadowaki T, et al.Role for gingipains in Porphyromonas gingivalis traffic to phagolysosomes and survival in human aortic endothelial cells[J]. Infect Immun, 2007, 75(5):2090-2100.
[4]	Genco CA, Kapczynski DR, Cutler CW, et al.Influence of immunization on Porphyromonas gingivalis colonization and invasion in the mouse chamber model[J]. Infect Immun, 1992, 60(4):1447-1454.
[5]	Ebersole JL, Brunsvold M, Steffensen B, et al.Effects of immunization with Porphyromonas gingivalis and Prevo-tella intermedia on progression of ligature-induced perio-dontitis in the nonhuman primate Macaca fascicularis[J]. Infect Immun, 1991, 59(10):3351-3359.
[6]	Gibson FC 3rd, Genco CA. Prevention of Porphyromonas gingivalis-induced oral bone loss following immunization with gingipain R1[J]. Infect Immun, 2001, 69(12):7959-7963.
[7]	Miyachi K, Ishihara K, Kimizuka R, et al.Arg-gingipain A DNA vaccine prevents alveolar bone loss in mice[J]. J Dent Res, 2007, 86(5):446-450.
[8]	Fayad R, Zhang H, Quinn D, et al.Oral administration with papillomavirus pseudovirus encoding IL-2 fully restores mucosal and systemic immune responses to vaccinations in aged mice[J]. J Immunol, 2004, 173(4):2692-2698.
[9]	Yonezawa H, Ishihara K, Oluda K.Arg-gingipain a DNA vaccine induces protective immunity against infection by Porphromonas gingivalis in a murine model[J]. Infect Im-mun, 2001, 69(5):2858-2864.
[10]	Chen T, Duncan MJ.Gingipainadhesin domains mediate Porphyromonas gingivalis adherence to epithelial cells[J]. Microb Pathog, 2004, 36(4):205-209.
[11]	Nogueira-Paiva NC, Fonseca Kda S, Vieira PM, et al.Myent-eric plexus is differentially affected by infection with distinct Trypanosoma cruzi strains in Beagle dogs[J]. Mem Inst Os-waldo Cruz, 2014, 109(1):51-60.
[12]	Rybak ER, Shipley S, Tatarov I, et al.Clinical Trypanosoma cruzi disease after cardiac transplantation in a cynomolgus macaque (Macaca fascicularis)[J]. Comp Med, 2016, 66(6):494-498.
[13]	Tokuda M, Karunakaran T, Duncan M, et al.Role of Arg-gingipain A in virulence of Porphyromonas gingivalis[J]. Infect Immun, 1998, 66(3):1159-1166.
[14]	Moritz AJ, Cappelli D, Lantz MS, et al.Immunization with Porphyromonas gingivalis cysteine protease: effects on experimental gingivitis and ligature-induced periodontitis in Macaca fascicularis[J]. J Periodontol, 1998, 69(6):686-697.
[15]	Iglesias E, Aguilar JC, Cruz LJ, et al.Broader cross-reactivity after conjugation of V3 based multiple antigen peptides to HBsAg[J]. Mol Immunol, 2005, 42(1):99-104.

组别	2周	4周	6周
A₁	2.13±0.09	3.47±0.12	4.32±0.73
A₂	0.66±0.18	0.82±0.26	1.02±0.37
B₁	2.51±0.11	3.78±0.11	4.86±0.36
B₂	0.72±0.23	0.89±0.34	1.37±0.48
C₁	3.58±0.99	5.12±0.09	5.37±0.41
C₂	0.79±0.28	0.97±0.27	1.46±0.52

组别	免疫前		免疫后
组别	sIgA	IgG	sIgA	IgG
A	2.57±0.09	2.37±0.12	4.32±0.73*	5.00±0.42*
B	2.51±0.11	2.41±0.12	4.15±0.36*	2.39±0.28*
C	2.48±0.99	2.33±0.09	2.27±0.41	2.53±0.46

牙龈卟啉单胞菌精氨酸特异性牙龈素基因疫苗预防比格犬种植体周围炎的实验研究

Experimental research on Arginine-gingipain A gene vaccine from Porphyromonas gingivalis that prevents peri-implantitis in Beagle dogs

RichHTML

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 15

相关文章 15

编辑推荐

Metrics

本文评价

[1]	王良涛, 李珊, 陆豆豆, 陈铮. 基于正交试验梯度多孔牙种植体结构的设计研究[J]. 华西口腔医学杂志, 2023, 41(6): 647-652.
[2]	郭力嘉, 刘奕彤, 陈颖怡, 徐骏疾, 刘怡. 牙龈卟啉单胞菌来源的脂多糖诱导非酒精性脂肪肝的局部巨噬细胞免疫耐受表型[J]. 华西口腔医学杂志, 2023, 41(4): 385-394.
[3]	苏文祺, 李静雯, 蒋立姗, 崔雯洁, 赵阳, 李厚轩. 口腔显微镜辅助种植体表面去污的体外研究[J]. 华西口腔医学杂志, 2023, 41(3): 350-355.
[4]	张雨薇, 何雨轩, 丁一, 刘程程. 牙龈卟啉单胞菌上调钙结合蛋白1促进牙龈上皮细胞增殖[J]. 华西口腔医学杂志, 2022, 40(1): 93-99.
[5]	尧可, 蔡靖仪, 赵蕾, 吴亚菲, 赵志河, 申道南. 牙龈卟啉单胞菌双组分信号转导系统的研究进展[J]. 华西口腔医学杂志, 2021, 39(1): 88-93.
[6]	周洁, 甘雪琦, 卢嘉仪, 范林莉, 朱卓立. 牙种植体作支抗局部正畸直立近中倾斜下颌第二磨牙1例[J]. 华西口腔医学杂志, 2020, 38(6): 708-711.
[7]	宋应亮, 张思佳. 对种植体周围炎的认识与预防[J]. 华西口腔医学杂志, 2020, 38(5): 479-483.
[8]	王春萌,洪丽华,张志民,王渝. 牙龈卟啉单胞菌在消化系统恶性肿瘤中的作用及机制[J]. 华西口腔医学杂志, 2019, 37(5): 521-526.
[9]	甘雪琦,肖宇,马瑞阳,黄椿棚,吴尧,杨帮成,杨齐,包崇云,于海洋. 牙种植体的生物力学研究[J]. 华西口腔医学杂志, 2019, 37(2): 115-123.
[10]	程磊,于海洋,吴尧,包崇云,杨帮成,满毅,孙瑶,岩晓丽,周学东. 牙种植体周围微生物研究[J]. 华西口腔医学杂志, 2019, 37(1): 7-12.
[11]	杨芸,陈珊珊,许春梅,吴亚菲,赵蕾. 牙龈卟啉单胞菌脂多糖刺激巨噬细胞表达髓样细胞触发受体-1的研究[J]. 华西口腔医学杂志, 2018, 36(5): 475-481.
[12]	容明灯, 黄雁红, 卢海宾, 徐格林, 李少冰, 苏媛, 陈沛, 姜盼, 张雪洋. 显微牙周外科技术在种植体周附着龈增宽术中的应用[J]. 华西口腔医学杂志, 2018, 36(1): 71-75.
[13]	赵妍, 于阳, 寇育荣. 牙龈卟啉单胞菌调控细胞自噬的分子机制研究进展[J]. 华西口腔医学杂志, 2017, 35(6): 654-658.
[14]	张尽美, 赵蕾, 吴亚菲. 牙龈卟啉单胞菌第Ⅸ型分泌系统的研究进展[J]. 华西口腔医学杂志, 2017, 35(5): 538-542.
[15]	陈筑, 宿凌恺. 丹皮酚对牙龈卟啉单胞菌诱导骨髓来源巨噬细胞功能的影响[J]. 华西口腔医学杂志, 2017, 35(2): 139-144.