Journal: International Journal of Molecular Sciences

Article Title: Annulus Fibrosus Injury Induces Acute Neuroinflammation and Chronic Glial Response in Dorsal Root Ganglion and Spinal Cord—An In Vivo Rat Discogenic Pain Model

doi: 10.3390/ijms25031762

Figure Lengend Snippet: AF puncture injury increased the level of CD68 in the spine. ( A ) Representative images of CD68 IHC staining with toluidine blue in the entire IVD, anterior AF (yellow box), anterior longitudinal ligament (green box), and endplate (red box) at 3 days and 8 weeks after AF injury. The highlighted regions (black box) in anterior AF, anterior longitudinal ligament, and endplate show a closer look for positive, brown CD68 staining (indicated with arrow) vs. negative blue staining in each group from naïve to 8 weeks. ( B ) Semi-quantitative analysis showed that the CD68-ir in anterior AF increased at 3 days post-injury and remained elevated until 8 weeks post-injury, while the CD68-ir in the anterior longitudinal ligament and endplate were temporary, with peaks at 1 week and 3 days, respectively, and then significantly decreased. * p < 0.05. ** p < 0.01. *** p < 0.001. **** p < 0.0001. Naïve (n = 4), 3 days (n = 6), 1 week (n = 7), 2 weeks (n = 4), 8 weeks (n = 7). The connecting line is drawn to visualize mean changes in the measurements with time, and we note that each bar involves a separate experimental group sacrificed at that time point, and that each dot represents a single animal.

Article Snippet: After antigen retrieval with IHC Enzo (ADI950-280-0015, ENZO Life Science, Farmingdale, NY, USA) and protein block (X0909, Agilent Technologies, Inc., Santa Clara, CA, USA), the sections were incubated for 1 h at room temperature with rabbit monoclonal primary antibody against rat CD68 (1:500 dilution, ab283654, Abcam, Waltham, MA, USA), or normal rabbit serum (S-500-20, Vector Laboratories, Inc., Burlingame, CA, USA) as the negative control.

Techniques: Immunohistochemistry, Staining

Journal: International Journal of Molecular Sciences

Article Title: Annulus Fibrosus Injury Induces Acute Neuroinflammation and Chronic Glial Response in Dorsal Root Ganglion and Spinal Cord—An In Vivo Rat Discogenic Pain Model

doi: 10.3390/ijms25031762

Figure Lengend Snippet: Temporal crosstalk of inflammatory responses between spine, DRG, and SC at acute and chronic states following AF injury. AF injury induced increased spine CD68-ir (anterior AF, purple line) and DRG Iba1-ir (red line) with peak at 3 days post-injury, followed by SC Iba1-ir (blue line) with a peak at 2 weeks post-injury.

Article Snippet: After antigen retrieval with IHC Enzo (ADI950-280-0015, ENZO Life Science, Farmingdale, NY, USA) and protein block (X0909, Agilent Technologies, Inc., Santa Clara, CA, USA), the sections were incubated for 1 h at room temperature with rabbit monoclonal primary antibody against rat CD68 (1:500 dilution, ab283654, Abcam, Waltham, MA, USA), or normal rabbit serum (S-500-20, Vector Laboratories, Inc., Burlingame, CA, USA) as the negative control.

Techniques: