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Fig 1.

Study design.

Open boxes represent datasets. In the screening cohort, nine patients with MRONJ stage ≥1 and nine patients before treatment with BMAs were compared by metabolomic analysis. In the validation cohort, eight patients with MRONJ stage ≥1 and nine patients without MRONJ after treatment with BMAs were compared in order to confirm the validity of findings in the screening cohort in a more clinically relevant setting.

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Fig 1 Expand

Fig 2.

Sample collection.

Patients provided a saliva sample following overnight fasting (~0.5 mL; minimum: 0.1 mL), transferred the sample to a cold-storage bag, and transported to the hospital within 4 h of collection, where it was stored at −80°C until use.

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Fig 3.

Heat map of salivary metabolomic profiles.

Nine saliva samples from patients before BMA and nine from patients with MRONJ were analyzed by CE-MS. Of the 232 metabolites analyzed, 118 were detected in ≥50% of samples and used for supervised cluster analysis to separate the two groups. Color codes: red, high; blue, low; and white, average.

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Fig 4.

Candidate metabolites in the two cohorts.

Five metabolites, including histamine, 3-(4-hydroxyphenyl)propionate, malonate, carnosine, and hypotaurine, showed the highest concentrations in the MRONJ group in the (A) screening cohort. In the (B) validation cohort, hypotaurine [MRONJ(−) vs MRONJ(+), 9.4 ± 11.2 × 10−4 vs. 26.6 ± 14.2 × 10−4; p = 0.017] and malonate (8.9 ± 5.6 × 10−4 vs. 21.3 ± 45.8 × 10−4; p = 0.472) showed consistently higher concentrations in MRONJ patients, although only hypotaurine was significantly higher. *p < 0.05.

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Fig 4 Expand

Table 1.

Patient characteristics.

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Table 1 Expand

Fig 5.

Relationships between hypotaurine concentration and MRONJ stage.

Hypotaurine concentrations were analyzed in specimens from the screening and validation patient cohorts. The graph shows data representing the mean ± SD. The open and filled bar graphs show MRONJ(−) and MORONJ(+), respectively. There was a significant difference in hypotaurine concentration between the control (n = 18) and stage 2 groups (n = 9) but not with the stage 1 group (n = 6) or stage 3 group (n = 2). *p < 0.05.

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Fig 5 Expand