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Journal of Peking University (Health Sciences) ›› 2023, Vol. 55 ›› Issue (1): 22-29. doi: 10.19723/j.issn.1671-167X.2023.01.004

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Comparison of three methods for establishing rat peri-implantitis model

Ling-wei MENG1,Xue LI2,Sheng-han GAO1,Yue LI1,Rui-tao CAO1,Yi ZHANG2,*(),Shao-xia PAN1,*()   

  1. 1. Department of Prosthodontics, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digi-tal Medical Devices & Beijing Key Laboratory of Digital Stomatology & NHC Research Center of Engineering and Technology for Computerized Dentistry & NMPA Key Laboratory for Dental Materials, Beijing 100081, China
    2. Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China
  • Received:2022-10-11 Online:2023-02-18 Published:2023-01-31
  • Contact: Yi ZHANG,Shao-xia PAN E-mail:zhangyi612@hotmail.com;panshaoxia@vip.163.com
  • Supported by:
    the National Key R & D Program of China(2020YFC2009005);National Program for Multidisciplinary Cooperative Treatment on Major Diseases(PKUSSNMP-202004);Beijing Natural Science Foundation(7222228)

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Abstract:

Objective: To compare the efficiency and effect of establishing rat peri-implantitis model by traditional cotton thread ligation and local injection of Porphyromonas gingivalis lipopolysaccharide (LPS) around the implant, as well as the combination of the two methods. Methods: Left side maxillary first molars of 39 male SD rats were extracted, and titanium implants were implanted after four weeks of healing. After 4 weeks of implant osseointegration, 39 rats were randomly divided into 4 groups. Cotton thread ligation (n=12), local injection of LPS around the implant (n=12), and the two methods combined (n=12) were used to induce peri-implantitis, the rest 3 rats were untreated as control group. All procedures were conducted under 5% isoflurane inhalation anesthesia. The rats were sacrificed 2 weeks and 4 weeks after induction through carbon dioxide asphyxiation method. The maxilla of the rats in the test groups were collected and marginal bone loss was observed by micro-CT. The gingival tissues around the implants were collected for further real time quantitative PCR (RT-qPCR) analysis, specifically the expression of tumor necrosis factor-alpha (TNF-α) as well as interleukin-1β (IL-1β). The probing depth (PD), bleeding on probing (BOP) and gingival index (GI) of each rat in the experimental group were recorded before induction of inflammation and before death. Results: After 4 weeks of implantation, the osseointegration of implants were confirmed. All the three test groups showed red and swollen gums, obvious marginal bone loss around implants. After 2 weeks and 4 weeks of inflammation induction, PD, GI and BOP of the three test groups increased compared with those before induction, but only BOP was statistically significant among the three test groups (P < 0.05). At the end of 2 weeks of inflammation induction, marginal bone loss was observed at each site in the cotton thread ligation group and the combined group. At each site, the bone resorption in the combined group was greater than that in the cotton thread ligation group, but the difference was not statistically significant (P > 0.05), bone resorption was observed at some sites of some implants in LPS local injection group. At the end of 4 weeks of inflammation induction, marginal bone loss was observed at all sites in each group. The marginal bone loss in the cotton thread ligation group and the combined group was greater than that in the LPS local injection group, and the difference was statistically significant (P < 0.05). At the end of 2 weeks and 4 weeks of induction, the expression of TNF-α and IL-1β in the test groups were higher than those in the control group (P < 0.05). Conclusion: Compared with local injection of LPS around the implant, cotton thread ligature and the two methods combined can induce peri-implantitis in rats better and faster.

Key words: Peri-implantitis, Animal models, Rats, Cotton thread ligation, Lipopolysaccharide local injection

CLC Number: 

  • R781.42

Figure 1

Tooth extraction, implant surgery and induction of peri-implantitis A, extraction of maxillary first molars; B, implantation preparation; C, implant insertion and suture; D, ligarue around the implant."

Table 1

The primer sequences of different genes for RT-qPCR"

Gene Primer sequences
TNF-α Forward: 5′-CTTCTCATTCCTGCTCGTG-3′
Reverse: 5′-TTTGGGAACTTCTCCTCCT-3′
IL-1β Forward: 5′-TTCATCTTTGAAGAAGAGCCC-3′
Reverse: 5′-CTGTCTAATGGGAACATCACAC-3′
GAPDH Forward: 5′-AACTCCCATTCTTCCACCT-3′
Reverse: 5′-TTGTCATACCAGGAAATGAGC-3′

Table 2

PD, GI and BOP at baseline, 2 weeks, and 4 weeks of different induction methods (n=6)"

Time point Group Items PD/mm, ${\bar x}$±s GI BOP+/%
Medial Distal Palatal Buccal
2 weeks LPS Baseline 0.75±0.31 0.42±0.15 0.92±0.45 0.50±0.18 0.67±0.21 0
2 weeks 1.50±0.48 0.83±0.28 1.58±0.52 1.08±0.44 1.33±0.42 20.8
P 0.220 0.220 0.359 0.246 0.240 0.025
Ligation Baseline 2.58±0.33 1.75±0.36 1.67±0.42 2.08±0.37 0.33±0.21 0
2 weeks 3.00±0.26 2.83±0.31 2.67±0.21 3.00±0.26 2.00±0.00 50.0
P 0.341 0.045 0.060 0.072 0.002 < 0.001
LPS+Ligation Baseline 2.50±0.34 2.25±0.40 1.75±0.44 2.17±0.30 0.17±0.17 0
2 weeks 2.91±0.20 2.75±0.25 2.83±0.21 2.92±0.27 2.00±0.00 83.3
P 0.318 0.317 0.052 0.097 0.002 < 0.001
4 weeks LPS Baseline 1.67±0.48 0.75±0.48 1.08±0.47 1.00±0.43 0.67±0.21 0
4 weeks 2.17±0.38 1.33±0.42 2.00±0.48 1.67±0.44 1.83±0.17 58.3
P 0.432 0.382 0.205 0.304 0.009 < 0.001
Ligation Baseline 1.75±0.36 1.58±0.37 2.08±0.45 2.00±0.37 0.50±0.22 0
4 weeks 2.83±0.28 2.91±0.20 2.92±0.20 3.00±0.13 2.17±0.17 91.7
P 0.039 0.011 0.125 0.027 0.002 < 0.001
LPS+Ligation Baseline 2.17±0.31 2.00±0.45 1.67±0.42 2.17±0.40 0.17±0.17 0
4 weeks 2.92±0.20 3.08±0.27 3.17±0.17 3.17±0.28 2.33±0.21 95.8
P 0.068 0.065 0.008 0.068 0.002 < 0.001

Figure 2

Micro-CT images regarding marginal bone loss after 2 weeks of induction LPS, local injection of Porphyromonas gingivalis lipopolysaccharide; Ligation, cotton thread ligation."

Figure 3

The peri-implant marginal bone loss of different groups after 2 weeks of induction (n=6) LPS, local injection of Porphyromonas gingivalis lipopolysaccharide; Ligation, cotton thread ligation; .M, medial; D, distal; P, palatal; B, buccal; ns, no statistically significant difference. *P < 0.001."

Figure 4

Micro-CT images regarding marginal bone loss after 4 weeks of induction LPS, local injection of Porphyromonas gingivalis lipopolysaccharide; Ligation, cotton thread ligation."

Figure 5

The peri-implant marginal bone loss of different groups after 4 weeks of induction (n=6) LPS, local injection of Porphyromonas gingivalis lipopolysaccharide; Ligation, cotton thread ligation; .M, medial; D, distal; P, palatal; B, buccal; ns, no statistically significant difference. *P < 0.001."

Figure 6

Relative TNF-α and IL-1β expression in peri-implant gingival tissure (n=3) TNF-α, tumor necrosis factor-α; IL-1β, interleukin-1β; LPS, local injection of Porphyromonas gingivalis lipopolysaccharide; Ligation, cotton thread ligation. *P < 0.05, vs. control; #P < 0.05, vs. same group at 2 weeks."

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