Journal: Bioactive Materials

Article Title: Nanomaterial-assisted theranosis of bone diseases

doi: 10.1016/j.bioactmat.2022.12.014

Figure Lengend Snippet: Nanocrystal-polymer and chitosan-based nanocarriers for osteoarthritis therapy. A) Schematic representation of a KGN-NPP. (i) SEM of surface and core of KGN-NPPs. (ii) Representative safranin-O/fast green and IHC showing the medial side of the knee of mice at day 56. Femur (above), tibia (below). Arrows indicate cartilage erosion. Scale bar = 200 μm . The images reproduced with the permission from WILEY-VCH. B) Illustration of the procedures to synthesize PN-KGN. (iii) TEM of PN-KGN. (iv) In vitro release of KGN from PN-KGN at 37 °C. (v) Macroscopic views of OA development at 3, 6, and 12 weeks after OA induction. Arrows indicate the locations of cartilage degeneration (n = 6 per group) . The images reproduced with the permission from Taylor & Francis. C). Proposed reaction scheme of CHI-KGN conjugate. (vi) Safranin-O and Alcian-blue staining on day 21. (vii) In vivo fluorescence imaging and relative intensity using fluorescence dye-labeled CHI-KGN NPs (left knee) and CHI-KGN MPs (right knee) at various time points from rats in which OA was induced surgically 6 weeks before . The images reproduced with the permission from Elsevier Ltd. D) Effects of BBR-loaded CNs on protein expressions of caspase-3, Bcl-2 and Bax in rat articular cartilage from each experimental group. (viii) Effect of BBR-loaded CNs on histological changes in articular cartilage of the rat ACLT + MMx model . The images reproduced with the permission from Elsevier B.V. E) SEM of C-CP-Fe-bLf-NCs and chemical bonding analysed using FTIR analysis. (ix) Fluorescence and biodistribution of arthritic mice treated with (a) control diet and (b&c) Cy5.5 dye labeled AEC-CP-Fe-bLf-NC diet were fed orally 36 h prior to being euthanized and imaged. Arrows show the localization of NC in joint cartilage. (x) Schematic diagram illustrating the signaling mechanism modulated by AEC-CP-Fe-bLf-NCs in arthritis . The images reproduced with the permission from Elsevier Ltd.

Article Snippet: KGN coated with chitosan NPs using EDC/NHS catalysis methods (CHI-KGN) showed a distinct inhibitory effect on cartilage degeneration ( C) [ ].

Techniques: Polymer, In Vitro, Staining, In Vivo, Fluorescence, Imaging, Labeling, Control

Journal: The FASEB Journal

Article Title: Timely expression and activation of YAP1 in granulosa cells is essential for ovarian follicle development

doi: 10.1096/fj.201900179RR

Figure Lengend Snippet: YAP1 interacts with EGFR signaling pathway to drive granulosa cell proliferation. A) Knockdown of Yap1 in mouse granulosa cells reduced mRNA levels of Egfr, EGF-like ligands (Hbegf and Areg), and cyclin D1 (Ccnd1). Relative mRNA levels were determined using RT-PCR. The experiment was repeated ≥3 times, and representative images are presented. B) Expression of HBEGF in hGCs, KGN cells, and HGrC1 cells transfected with empty vectors [MX as control (CTL)], vectors expressing wild-type YAP1 (YAP), or constitutively active YAP1 (YAPS127A). Relative mRNA levels were quantified using real-time PCR. Each bar represents the mean ± sem (n = 4). C, D) HBEGF treatment (50 ng/ml, 3 d) stimulated cell proliferation (C), but suppressed estradiol (E2) and progesterone (P4) production (D) in cultured granulosa cells. Each bar represents the mean ± sem (n = 4). E) HBEGF (50 ng/ml) induced rapid dephosphorylation of LATS1 (S909) and YAP1 (S127) and phosphorylation of ERK1/2 in cultured granulosa cells. β-Actin protein was used as loading CTL. The relative levels of the total proteins and phosphoproteins were determined by Western blot. Ctgf, connective tissue growth factor; Gapdh, glyceraldehyde 3-phosphate dehydrogenase; p, phosphorylated. Experiments were repeated ≥3 times, and representative images are presented. *P < 0.05, **P < 0.01, ***P < 0.001, compared with corresponding CTL groups (MX groups).

Article Snippet: HGrC1 [an immortalized human granulosa cell (hGC) line] and KGN (an ovarian granulosa cell–like tumor cell line) cells were obtained from the Riken Cell Bank (Riken BioResource Research Center, Tsukuba, Japan).

Techniques: Knockdown, Reverse Transcription Polymerase Chain Reaction, Expressing, Transfection, Control, Real-time Polymerase Chain Reaction, Cell Culture, De-Phosphorylation Assay, Phospho-proteomics, Western Blot

Journal: The FASEB Journal

Article Title: Timely expression and activation of YAP1 in granulosa cells is essential for ovarian follicle development

doi: 10.1096/fj.201900179RR

Figure Lengend Snippet: YAP1 stimulates proliferation but suppresses differentiation of granulosa cells. A) Representative images showing spheroids formed in KGN-MX, KGN-YAP, and KGN-YAPS127A cells in a 3D culture system. Scale bars, 200 μm. B) Numbers of spheroids with different diameters formed by KGN-MX, KGN-YAP, and KGN-YAPS127A cells in the 3D culture system in A. Each bar represents the mean ± sem (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001 compared with corresponding control (CTL). C) Production of E2 and progesterone in KGN-MX, KGN-YAP, and KGN-YAPS127A cells in the 3D culture system. *P < 0.05, **P < 0.01, ***P < 0.001 compared with MX control. D) Knockdown of Yap1 with Yap1-specific siRNA (siYap) in cultured mouse granulosa cells up-regulated the expression of granulosa cell differentiation–associated genes. The relative mRNA levels of Star, Fshr, Sult1e1, Ptgfr, cyp11A1, cyp19a1, and Gapdh were determined by RT-PCR. E) Forskolin (FSK) induced expression of differentiation-associated genes in cultured primary mouse granulosa cells. The relative mRNA levels of Yap1, Star, Fshr, Sult1e1, Ptgfr, cyp11a1, and cyp19a1 in mouse granulosa cells treated with 10 μM FSK for the indicated time points were determined by quantitative PCR. *P < 0.05, **P < 0.01, ***P < 0.001 compared with CTL. F) FSK (10 μM) induced phosphorylation of YAP1 in cultured HGrC1 cells. Please note that phosphorylation results in inactivation of YAP1 protein. The relative levels of the total proteins and phosphoproteins were determined by Western blot. All experiments were repeated ≥3 times, and representative images were presented. G) FSK (10 μM) suppressed the expression of Hbegf mRNA in cultured primary mouse granulosa cells in a time-dependent manner. Each bar represents the mean ± sem (n = 4). *P < 0.05, **P < 0.01 compared with CTL. H) FSK (10 μM, 72 h) suppressed HBEGF mRNA expression in cultured primary hGCs. CREB, cAMP responsive element binding protein; Gapdh, glyceraldehyde 3-phosphate dehydrogenase; p, phosphorylated. Each bar represents the mean ± sem (n = 4). **P < 0.01 compared with CTL.

Article Snippet: HGrC1 [an immortalized human granulosa cell (hGC) line] and KGN (an ovarian granulosa cell–like tumor cell line) cells were obtained from the Riken Cell Bank (Riken BioResource Research Center, Tsukuba, Japan).

Techniques: Control, Knockdown, Cell Culture, Expressing, Cell Differentiation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Phospho-proteomics, Western Blot, Binding Assay

Journal: Bioengineering & Translational Medicine

Article Title: Kartogenin (KGN)/synthetic melanin nanoparticles (SMNP) loaded theranostic hydrogel scaffold system for multiparametric magnetic resonance imaging guided cartilage regeneration

doi: 10.1002/btm2.10364

Figure Lengend Snippet: Schematic illustration of preparation and functionalization of kartogenin loaded and SMNP labeled multifunctional hydrogel scaffold for cartilage regeneration

Article Snippet: Kartogenin (KGN) and acetone were obtained from Macklin Co., Ltd. (Shanghai, China).

Techniques: Labeling