Fig 1.
The study population (A), sampling (B), and methods (C) are summarized in this figure. The study included thirty Bothrops snakebite patients, categorized into Mild and Severe groups based on clinical severity classification, alongside a Healthy Donor (HD) control group. Circulating levels of Matrix Metalloproteinases (MMPs) and their respective Tissue Inhibitors (TIMPs) were quantified before (T0) and post-antivenom therapy (T1 and T2). Created in BioRender. Gama, F. (2026) https://BioRender.com/new56x5.
Table 1.
Sociodemographic and clinical characterization of the study population.
Fig 2.
Profile of metalloproteinases and metalloproteinase inhibitors in Bothrops snakebite patients before the administration of antivenom.
The patients were divided into Mild (), Severe (
), and HD (
) groups. Circulating levels of Matrix Metalloproteinases (MMPs) and their respective Tissue Inhibitors (TIMPs) were quantified using a multiplex assay, as detailed in the Methods section. The results are presented using bar and symbol charts, reported in log10 scale, showing the median with IQR, expressed in picograms per milliliter (pg/mL). Statistical analysis was performed using the Kruskal-Wallis test followed by Dunn’s post-hoc test, with significant differences denoted by asterisks: *p < 0.05, **p < 0.01, and ***p < 0.001.
Fig 3.
Dynamics of the profile of metalloproteinases and metalloproteinase inhibitors in Bothrops snakebite patients after the administration of antivenom.
Circulating levels of Matrix Metalloproteinases (MMPs) and their respective Tissue Inhibitors (TIMPs) were quantified before (T0) and post-antivenom therapy (T1 and T2) in Group Mild () and Severe (
) groups. The interquartile range [25-75] of molecule concentrations in the Healthy Donor (HD) control group was used as baseline (
). The results are presented using bar and symbol charts, reported in log10 scale, showing the median with IQR, expressed in picograms per milliliter (pg/mL). Statistical analyses were performed using the Wilcoxon test for comparisons between time points (T0, T1, and T2) and the Mann-Whitney U test for inter-group comparisons. Statistically significant differences (p < 0.05) between follow-up days are represented by the dash symbol (-). Differences compared to the HD group are highlighted with asterisks (*). Significant differences between the Mild and Severe groups are represented by the hash symbol (#).
Fig 4.
Signature of metalloproteinases and metalloproteinase inhibitors in mild and severe patients.
The signature of Matrix Metalloproteinases (MMPs) and their respective Tissue Inhibitors (TIMPs) in Bothrops snakebite patients was constructed before (T0) and post-antivenom therapy (T1 and T2). The data, originally expressed in picograms per milliliter (pg/mL), were converted to categorical data using the overall median values as a cutoff to classify the study population as having low or high production of the MMPs and TIMPs evaluated. The overall signatures were assembled in radar charts using the 50th percentile as the threshold (central circle/gray zone) to identify MMPs and TIMPs with increased levels in a greater proportion of patients.
Fig 5.
Integrative networks of Bothrops snakebite patients in blood samples.
Networks of patients with bothropic snakebite were constructed before (T0) and post-antivenom therapy (T1 and T2) to identify the complex interactions among Matrix Metalloproteinases (MMPs) and their respective Tissue Inhibitors (TIMPs). The nodes () and (
) are used to identify the MMPs and TIMPs analyzed in the study. Solid lines between elements indicate a positive correlation, while dashed lines indicate a negative correlation. The thickness of the lines indicates the strength of the correlation. The correlation index (r) was used to categorize the strength of the correlation as weak (r ≤ 0.35), moderate (r ≥ 0.36 to r ≤ 0.67), or strong (r ≥ 0.68).
Fig 6.
Schematic summary of the local inflammatory response to Bothrops envenomation in Mild and Severe patients.
This model illustrates the Matrix Metalloproteinases (MMPs) and their respective Tissue Inhibitors (TIMPs) dynamics in Bothrops envenomation, from inoculation to tissue repair. (A) Inflammatory Response and Initial Activation (T0): Venom inoculation induces acute damage and the release of MMPs (MMP-1, -2, -7, -10) and TIMPs, establishing an initial proteolytic imbalance. (B) Severity Differentiation (T1): 24 hours after antivenom therapy, clinical progression is distinguished by enzymatic modulation. The Mild group exhibits efficient regulatory balance (TIMPs control MMPs), whereas the Severe group maintains a persistent imbalance (predominance of MMPs, such as MMP-9). (C) Resolution and Homeostasis (T2): In the Mild group, modulation is sufficient, with MMP-10 (an activator) and TIMPs promoting matrix extracellular (ECM) turnover and repair. The Severe group fails to reorganize the ECM, indicating dysfunctional and prolonged recovery. Created in BioRender. Gama, F. (2026) https://BioRender.com/ly48v07.