*2.3. CRO Reduced EAE-Induced Inflammatory Infiltrates in the Spinal Cord 28 Days after Immunization*

Central nervous system inflammation is a hallmark of MS pathogenesis [43]. Therefore, the effect of CRO in EAE-induced leukocyte migration to the spinal cord was investigated. The level of cellular infiltrate was scored from 0 (no cellular infiltrate) to 4 (high cellular

infiltrate) As shown in panel (Figure 3A), HE staining revealed that there was no cell infiltration in the control group treated with CFA, and the spinal cord of EAE mice displayed mixed cellular infiltrate with numerous perivascular clusters. Of interest, treatment with CRO significantly reduces the EAE-induced cell infiltration at the peak of the disease (Figure 3B), with no difference observed on the 28th day after immunization (Figure 3C).

**Figure 3.** A single dose of CRO reduces cellular infiltration in the spinal cord. The animals were sedated and euthanized for spinal cord collection. Representative 20× microscopic fields of transverse sections of the spinal cord stained with hematoxylin and eosin ((**A**), panel). Quantification of the inflammatory score (0–4) in 17th (**B**) and 28th (**C**) days. Data represent the mean ± SEM of 3 animals per group. \* *p* < 0.05 compared to the control group (CFA). # *p* < 0.05 compared to the untreated group (EAE). One-way ANOVA was used, followed by Tukey's post-hoc test.
