Table 1.
Oligonucleotide primers used in this study.
Table 2.
Recovery of viable P. gingivalis from oral swabs after final inoculation.
Figure 1.
Micro CT appearance and analysis of periodontal bone loss.
A. Micro CT of mouse jaw for CMC control group. B. Micro CT of mouse jaw for ECR527 & EA group. C. Micro CT of mouse jaw W50 & EA group. D. Measurements of bone changes in the jaw were calculated by measuring the cemento-enamel junction-alveolar bone crest on three slices for each mouse using CTAn sagital sections. Data represent mean (± SEM). The number of animals in each group was 6 and measurements were performed in triplicate. Statistical analysis was done by one way ANOVA and Tukey multiple comparisons. White bars represent CEJ/ABC distance measurements. Abbreviations: CMC = carboxymethyl cellulose; ECR527 = PAD-deficient P. gingivalis; W50 = wild type P. Gingivalis; EA = experimental arthritis
Figure 2.
Histological analysis of maxillary periodontal tissues.
A. Histology of mouse jaw for CMC control group at day 63. B. Histology of mouse jaw for ECR527 & EA group at day 63. C. Histology of mouse jaw for W50 & EA group at day 63. Arrow indicates increased inflammatory reaction in supra-crestal alveolar bone gingival tissue. D. Histological scores (mean ± SEM) for the presence of osteoclasts and evidence of bone erosion around the first and second upper molars for all groups in the study. Original magnification of A–C = 100X. The number of animals in each group was 6 and measurements were performed in triplicate. Statistical analysis was done by one way ANOVA and Tukey multiple comparisons. Abbreviations: CMC = carboxymethyl cellulose; ECR527 = PAD-deficient P. gingivalis; W50 = wild type P. Gingivalis; EA = experimental arthritis.
Figure 3.
Micro CT appearance and analysis of bone erosion in front paw radio-carpal joint.
A. Micro CT appearance of paw for CMC control group at day 63. B. Micro CT appearance of paw for ECR527 & EA group at day 63. C. Micro CT appearance of paw for W50 & EA group at day 63. D. The front paws (left and right combined), were processed to determine bone loss/growth between the two scanned time points in the radio-carpal joint. Data represent mean (± SEM). The number of animals in each group was 6 and measurements were performed in duplicate. Statistical analysis was done by one way ANOVA and Bonferroni multiple comparisons. Abbreviations: CMC = carboxymethyl cellulose; ECR527 = PAD-deficient P. gingivalis; W50 = wild type P. Gingivalis; EA = experimental arthritis
Figure 4.
Visual appearance and scoring of paw swelling.
A. Appearance of front and rear paws for CMC control group at day 7 post EA induction. B. Appearance of front and rear paws for ECR527 & EA group at day 7 post EA induction. C. Appearance of front and rear paws for W50 & EA group at day 7 post EA induction. D. Total paw score grade out of a 60 including front and back paws. Data represent mean (± SEM). The number of animals in each group was 6 and measurements were performed once daily per animal Statistical analysis was done by two way ANOVA and Tukey multiple comparisons. Raw data for data points are shown in Figure S1. Abbreviations: CMC = carboxymethyl cellulose; ECR527 = PAD-deficient P. gingivalis; W50 = wild type P. Gingivalis; EA = experimental arthritis
Figure 5.
Histological assessment of joint inflammation.
A. Histology of the radio-carpal joint for CMC control group at day 63. B. Histology of the radio-carpal joint for ECR527 & EA group at day 63. C. Histology of the radio-carpal joint for W50 & EA group at day 63. Arows indicate increased inflamaotry reaction in joint. D. Histological scores (mean ± SEM) for (a) inflammation and (b) bone and cartilage destruction for both front and rear paws combined. Original magnification of A–C = 100X. The number of animals in each group was 6 and measurements were performed in triplicate. Statistical analysis was done by One way ANOVA and Bonferroni multiple comparisons. Abbreviations: CMC = carboxymethyl cellulose; ECR527 = PAD-deficient P. gingivalis; W50 = wild type P. Gingivalis; EA = experimental arthritis
Figure 6.
Serum anti-CCP antibody levels.
The mean (± SEM) titre of anti-CCP antibodies detected in the sera of mice at week 9 from each of the 6 treatment groups. Raw data for data points are shown in Figure S2. The number in each group was 6 and measurements were performed in duplicate. Statistical analysis was done by one way ANOVA and Tukey multiple comparisons. Abbreviations: CMC = carboxymethyl cellulose; ECR527 = PAD-deficient P. gingivalis; W50 = wild type P. Gingivalis; EA = experimental arthritis
Figure 7.
Serum anti-P. gingivalis levels.
The mean (± SEM) titre of anti-P.gingivalis detected in the sera of mice over 9 week time course. Raw data for data points are shown in Figure S3. The number in each group was 6 and measurements were performed in duplicate. Statistical analysis was done by one way ANOVA and Tukey multiple comparisons. Abbreviations: CMC = carboxymethyl cellulose; ECR527 = PAD-deficient P. gingivalis; W50 = wild type P. Gingivalis; EA = experimental arthritis