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. 2019 May 29;9:8025. doi: 10.1038/s41598-019-44485-9

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Paw and Ankle Edema Modification by Ashwashila. (A) Normal control (NC) Balb/c mice showing digits (yellow arrow), foot (blue arrow) and ankle (red arrow). (B) Development of digits, foot and ankle edema in collagen antibody-induced arthritis (CAIA) disease control (DC) animal. (C) Reduction in the inflammation of digits, foot and ankle edema in CAIA mice treated with 353 mg/kg dose of Ashwashila (ASHW) every day for two weeks. (D) Reduction in inflammation of digits, foot and ankle edema in CAIA mice treated with 0.38 mg/kg dose of Methotrexate (MTX) every alternate day for two weeks (MTX). (E) Increase in paw edema of the observed in the CAIA animals. Treatment of the CAIA mice with ASHW or MTX induced significant reduction in the paw edema. (F) Percentage (%) activity of the ASHW or MTX treatments in reducing paw edema in the CAIA animals indicated similar efficacy. (G) Increase in ankle-joint edema was observed in the CAIA animals. Treatment with ASHW or MTX significantly reduced the ankle-joint edema in the CAIA animals. (H) Percentage (%) activity of the ASHW or MTX treatments in reducing knee-joint edema in the CAIA animals indicated similar efficacy. (I) Paw withdrawal threshold was measured using Randall Selitto (Mechanical hyperalgesia) parameter. The results showed a significant increase in the paw withdrawal threshold in the CAIA animals. Increase in mechanical hyperalgesia was recovered in the CAIA animals treated with Ashwashila (ASHW) and Methotrexate (MTX). (J) Thermal hyperalgesia test showed reduced in the latency time of CAIA animals followed by significant recovery when treated with ASHW and MTX. Values in the results are Mean ± SEM. A one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison t-test was used to calculate the statistical difference. Student unpaired t-test was used to calculate the statistical difference in comparison to MTX (p-value # ≤ 0.05; * ≤ 0.05).