Chuanhua Li, Zhifeng Wang, Lina Zhu, Xin Fan, Jing Lan
School of Stomatology, Shandong University, Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan 250012, China.
Dept. of Pediatrics Dentistry, School of Stomatology, Shandong University, Jinan 250012, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2018 Feb 1;36(1):76-81. doi: 10.7518/hxkq.2018.01.015.
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.
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.
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 significantly 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.
IgG and sIgA can be generated by immunity with rgpA DNA vaccine, which can significantly slow down bone loss during experimental peri-implantitis in dogs.
本研究旨在使用精氨酸牙龈蛋白酶A基因疫苗(pVAX1-rgpA)对成年比格犬进行免疫,并评估其在种植体周围炎进展和发展过程中的作用。
构建质粒pVAX1-rgpA。拔除15只成年比格犬双侧下颌第二、三前磨牙,并立即植入种植体。3个月后,将动物随机分为A、B、C组。之后,分别用质粒pVAX1-rgpA、热灭活牙龈卟啉单胞菌或pVAX1对动物进行三次免疫。免疫前及免疫2周后,采用酶联免疫吸附测定法定量分析血清中的IgG和唾液中的分泌型IgA(sIgA)。用棉线结扎种植体颈部诱导种植体周围炎。记录探诊深度(PD)和探诊出血情况。结扎6周后处死所有动物。制备50μm厚的脱钙切片,用亚甲蓝染色以观察种植体周围的骨表型。
免疫后A组和B组血清IgG水平及唾液中sIgA水平高于免疫前(P<0.05),且高于C组(P<0.05)。然而,A组和B组之间未观察到差异(P>0.05)。结扎后4周和6周,C组结扎侧的PD高于A组和B组(P<0.05)。另一方面,A组和B组之间未发现差异(P>0.05)。A组的骨吸收明显低于其他组(P<0.05)。通过硬组织切片观察发现,三组种植体周围骨吸收区域均存在大量炎性细胞和细菌。然而,C组种植体周围骨吸收区域的炎性细胞和细菌数量最多。
rgpA DNA疫苗免疫可产生IgG和sIgA,能显著减缓犬实验性种植体周围炎期间的骨吸收。