Micro CT appearance and analysis of bone erosion in front paw radio-carpal joint.

<p>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. <i>Abbreviations:</i> CMC = carboxymethyl cellulose; ECR527 = PAD-deficient <i>P. gingivalis</i>; W50 = wild type <i>P. Gingivalis</i>; EA = experimental arthritis</p

Histological analysis of maxillary periodontal tissues.

<p>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. <i>Abbreviations:</i> CMC = carboxymethyl cellulose; ECR527 = PAD-deficient <i>P. gingivalis</i>; W50 = wild type <i>P. Gingivalis</i>; EA = experimental arthritis.</p

Visual appearance and scoring of paw swelling.

<p>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 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0100838#pone.0100838.s001" target="_blank">Figure S1</a>. <i>Abbreviations:</i> CMC = carboxymethyl cellulose; ECR527 = PAD-deficient <i>P. gingivalis</i>; W50 = wild type <i>P. Gingivalis</i>; EA = experimental arthritis</p

Serum anti-CCP antibody levels.

<p>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 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0100838#pone.0100838.s002" target="_blank">Figure S2</a>. 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. <i>Abbreviations:</i> CMC = carboxymethyl cellulose; ECR527 = PAD-deficient <i>P. gingivalis</i>; W50 = wild type <i>P. Gingivalis</i>; EA = experimental arthritis</p

Oligonucleotide primers used in this study.

a<p>Underlined sequence of SOE primers indicates the part of the primer that is not complementary to the target sequence, but provides complementarity with a second PCR product for splicing.</p

Histological assessment of joint inflammation.

<p>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. <i>Abbreviations:</i> CMC = carboxymethyl cellulose; ECR527 = PAD-deficient <i>P. gingivalis</i>; W50 = wild type <i>P. Gingivalis</i>; EA = experimental arthritis</p

Serum anti-<i>P. gingivalis</i> levels.

<p>The mean (± SEM) titre of anti-<i>P.gingivalis</i> detected in the sera of mice over 9 week time course. Raw data for data points are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0100838#pone.0100838.s003" target="_blank">Figure S3</a>. 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. <i>Abbreviations:</i> CMC = carboxymethyl cellulose; ECR527 = PAD-deficient <i>P. gingivalis</i>; W50 = wild type <i>P. Gingivalis</i>; EA = experimental arthritis</p