Journal: Calcified tissue international

Article Title: A Mouse Noninvasive Intraarticular Tibial Plateau Compression Loading-Induced Injury Model of Posttraumatic Osteoarthritis

doi: 10.1007/s00223-019-00614-0

Figure Lengend Snippet: A: Longitudinal thin sections (5 μm) of the femoral-tibial junction, including the meniscus, were stained with alcian blue with orange G counterstain. GAG-proteoglycan of cartilage is stained in blue/purple color, and bone is stained in orange to red color. Arrows indicates the hypertropic chondrocyte areas in the injured meniscus. B: The hypertrophic chondrocyte and total tissue areas of the meniscus of both the injured right knee joint as well as the control contralateral intact knee joint were determined with the aid of an osteomeasure software. C: The comparison of the relative ratio of the hypertrophic chondrocyte area to total tissue area of meniscus of the injured joints with that of contralateral uninjured joint. Results are shown as mean±SEM, n=8 per group. *P<0.05; ***P<0.001, by two-tailed Student’s t-test.

Article Snippet: Accordingly, we drew a circle from the edge of the articular cartilage layer to the edge of the periarticular bone of the site-of-interest and then measured the total area of the circle using the OsteoMeasure system equipped with a digitizing tablet (Osteometrics, Atlanta, GA).

Techniques: Staining, Control, Software, Comparison, Two Tailed Test

Journal: bioRxiv

Article Title: Intermittent Fasting Mitigates Vascular and Neuronal Pathologies in a Mouse Model of Vascular Dementia

doi: 10.1101/2022.02.22.481534

Figure Lengend Snippet: A-F . Representative cresyl violet images and quantification illustrating Nissl positively stained neurons in hippocampal CA1, CA2 and CA3 regions. Magnification x60. Scale bar, 20 μm. Images were taken under identical exposures and conditions. Data are represented as mean ± standard error of the mean. n=5 mice in each experimental group. *P<0.05 compared with AL Sham; + P<0.05, ++ P<0.01, ++++ P<0.0001 compared with corresponding AL BCAS. G-L . Representative immunoblots and quantification illustrating apoptosis marker cleaved caspase-3 at the cortex (G-H) , hippocampus (I-J) and cerebellum (K-L) . Data are represented as mean ± standard error of the mean. n=5-7 mice in each experimental group. β-actin was used as a loading control. *P<0.05 compared with AL Sham; + P<0.05, ++ P<0.01 compared with corresponding AL group. Abbreviations: AL, ad libitum; BCAS, bilateral common carotid artery stenosis; Cl, cleaved; IF, intermittent fasting; VCI, vascular cognitive impairment.

Article Snippet: The nitrocellulose membranes were incubated with the following primary antibodies: VEGF (Santa Cruz, sc7269), PDGFRβ (Cell Signaling, #3169), ZO-1 (Thermo Fisher, 61-7300), Occludin (Invitrogen, 711500), Claudin-5 (Thermo Fisher, 35-2500), JAM-A (Santa Cruz, sc25629), MBP (Cell Signaling, #78896S), Cleaved Caspase-3 (Cell Signaling, #9664), Total Caspase-3 (Cell Signaling, #9662), MMP2 (Cell Signaling, #87809S), MMP9 (Millipore, AB19016), MT1-MMP (Cell Signaling, #13130S), Malondialdehyde (Abcam, ab6463), Superoxide Dismutase (Abcam, ab13498), Glutathione (Abcam, ab19534), Vinculin (Cell Signaling, #13901) and β-actin (Sigma-Aldrich, A5441).

Techniques: Staining, Western Blot, Marker

Journal: bioRxiv

Article Title: Intermittent Fasting Mitigates Vascular and Neuronal Pathologies in a Mouse Model of Vascular Dementia

doi: 10.1101/2022.02.22.481534

Figure Lengend Snippet: Vascular pathology in the brain following decreased cerebral blood flow (CBF) is defined as changes in the number of leaky microvessels and damage to the blood brain barrier (BBB). DiI staining was used to stain the vasculature, and IF was found to decrease the number of leaky microvessels in the brain following CCH. In addition, IF increased vascular growth factors in the brain, namely vascular endothelial growth factors (VEGF) and platelet derived growth factor receptor beta (PDGFRβ). Evans blue staining was used to assess the extent of BBB damage in the brain. IF was found to decrease BBB permeability through increased tight junction proteins zonula-occludens (ZO)-1, Occludin, Claudin-5 and junctional adhesion molecule (JAM)-A. Vascular pathology in the brain following decreased CBF is defined as changes in the white matter pathology and loss of hippocampal neurons. Luxol fast blue (LFB) staining method was used to stain and image 5 different white matter regions (corpus callosum medial, corpus callosum paramedian, caudoputamen, internal capsule and optic tract). IF decreased the severity of white matter damage in the brain following CCH, through maintenance of myelin basic protein (MBP) levels. Nissl staining was used to visualise the hippocampal neurons in the brain, and IF was found to be able to increase neuronal counts through reducing apoptotic death indicated by the levels of cleaved caspase-3 in the brain. IF was found to reduce membrane-type 1 MMP (MT1-MMP) and MMP2 levels in the brain following CCH. However, MMP9 was not involved in breaking down the extracellular matrix following CCH. IF reduced oxidative stress as evidenced through markers of lipid oxidation such as malondialdehyde, and antioxidative glutathione, and superoxide dismutase levels.

Article Snippet: The nitrocellulose membranes were incubated with the following primary antibodies: VEGF (Santa Cruz, sc7269), PDGFRβ (Cell Signaling, #3169), ZO-1 (Thermo Fisher, 61-7300), Occludin (Invitrogen, 711500), Claudin-5 (Thermo Fisher, 35-2500), JAM-A (Santa Cruz, sc25629), MBP (Cell Signaling, #78896S), Cleaved Caspase-3 (Cell Signaling, #9664), Total Caspase-3 (Cell Signaling, #9662), MMP2 (Cell Signaling, #87809S), MMP9 (Millipore, AB19016), MT1-MMP (Cell Signaling, #13130S), Malondialdehyde (Abcam, ab6463), Superoxide Dismutase (Abcam, ab13498), Glutathione (Abcam, ab19534), Vinculin (Cell Signaling, #13901) and β-actin (Sigma-Aldrich, A5441).

Techniques: Staining, Derivative Assay, Permeability