Journal: bioRxiv

Article Title: Cardiac fibroblasts regulate cardiomyocyte hypertrophy through dynamic regulation of type I collagen

doi: 10.1101/2022.05.25.493406

Figure Lengend Snippet: (A) Whole ventricle mRNA microarray analysis of Col1a2 -/- mouse hearts compared to Col1a2 +/- at 2 months of age, n=3 per genotype. (B) Mass spectrometry analysis of ECM protein changes in Col1a2 -/- mouse hearts compared to Col1a2 +/- hearts at 3 months of age, n=4 per genotype. (C) Representative immunofluorescence images and (D) Western blot analysis of periostin from hearts of Col1a2 +/- and Col1a2 -/- mice at 3 months of age. Scale bar: 25 µm. (E) Flow cytometric gate strategy and (F) analysis of cardiac fibroblasts (MEFSK4 + /CD31 - /CD45 - ) from dissociated hearts of Col1a2 +/- and Col1a2 -/- mice at 3 months of age. (G) Representative immunofluorescence images of platelet-derived growth factor receptor (PDGFR)-α (purple) in Col1a2 +/- and Col1a2 -/- mice at 3 months of age. Wheat germ agglutinin (WGA) staining is green and shows outlines of cardiomyocytes. Scale bar: 100 µm. Relative mRNA expression of Col1a2 (H), Postn (I), Col3a1 (J) and Col5a1 (K) in sorted cardiac fibroblasts (MEFSK4 + /CD31 - /CD45 - ) from Col1a2 +/- and Col1a2 -/- mice at 9 months of age. Student t -test for panels (F), (H), (I), (J) and (K).

Article Snippet: Antibodies against the following proteins were used: periostin (Novus Biologicals NBP1-30042; 1:300 dilution for IF, 1:1000 for Western blot); collagen I (Abcam ab21286; 1:100 for IF); PDGFRα from (R&D Systems AF1062; 1:1000 for IF); collagen 1a2 (Santa Cruz sc-393573; 1:500 for Western blot) Anti-CD31 was from BioLegend (102423; 1:100 for flow cytometry); anti-CD45 was from BD Biosciences (563890; 1:100 for flow cytometry); anti-MEFSK4 was from Miltenyi Biotec (130-120-802; used 1:30 for flow cytometry).

Techniques: Microarray, Mass Spectrometry, Immunofluorescence, Western Blot, Derivative Assay, Staining, Expressing

Journal: Advanced Science

Article Title: A 3D‐Bioprinted Functional Module Based on Decellularized Extracellular Matrix Bioink for Periodontal Regeneration

doi: 10.1002/advs.202205041

Figure Lengend Snippet: The GelMA/dECM bioink provided a favorable inductive microenvironment for DFCs. A) Live/dead staining and B) cell viability analysis of DFCs in the GelMA/dECM bioink for 7 days (* p = 0.0188, *** p = 0.0002, *** p = 0.0003, * p = 0.0421, ** p = 0.0014, *** p = 7.55 × 10 −9 , * p = 0.0427, ** p = 0.0012, ** p = 0.0063, n = 6 fields from three independent samples). C) Cell proliferation of DFCs in GelMA/dECM hydrogels determined via a CCK‐8 quantitative analysis after 7 days of culture (* p = 0.0297, NS = 0.1788, 0.5635, 0.4091). D) Evaluation of the chemotactic activities of the GelMA and GelMA/dECM bioink diluent on DFCs by transwell assay. E) Quantitative statistics of the positive area of DFCs that migrated to the opposite side of the membrane (NS = 0.9416, *** p = 6.2098 × 10 −7 , *** p = 1.383 × 10 −12 , n = 6 fields from three independent experiments). F) Real‐time PCR detection of osteogenic (Runx2, ALP, OPN, COL‐1) and odontogenic (DSPP, periostin, laminin‐ β 1) gene expression in DFCs cultured in the four hydrogels for 7 days (left: * p = 0.0351, * p = 0.0478, * p = 0.0267, * p = 0.0109; right: * p = 0.0203, * p = 0.0297, ** p = 0.0098, ** p = 0.0012, ** p = 0.0042, ** p = 0.0028, n = 3 independent replicates from three experiments). Scale bars = 200 µm. (Two‐sided comparison, error bars represent standard deviation, * p < 0.05, ** p < 0.01 and *** p < 0.001).

Article Snippet: The antibodies used for IF included periostin (1:300 dilution, Santa Cruz, USA), mitochondria (1:400 dilution, Abcam, UK), OCN (1:300 dilution, Zenbio, China), and OPG (1:200 dilution, Abcam, UK).

Techniques: Staining, CCK-8 Assay, Transwell Assay, Membrane, Real-time Polymerase Chain Reaction, Gene Expression, Cell Culture, Comparison, Standard Deviation

Journal: Advanced Science

Article Title: A 3D‐Bioprinted Functional Module Based on Decellularized Extracellular Matrix Bioink for Periodontal Regeneration

doi: 10.1002/advs.202205041

Figure Lengend Snippet: Periodontal regeneration induced by the 3D bioprinted periodontal module. A) H&E and Masson's trichrome staining showed evident new bone and PDL formation in the GelMA+5dECM group (black rectangle: defect area). B) IF staining of the PDL‐specific protein periostin and human cell‐specific protein mitochondria in newly formed tissues (white dotted lines: the bone–ligament and ligament–root interfaces). Scale bars = 100 µm. AB: alveolar bone; NB: new bone; PDL: periodontal ligament.

Article Snippet: The antibodies used for IF included periostin (1:300 dilution, Santa Cruz, USA), mitochondria (1:400 dilution, Abcam, UK), OCN (1:300 dilution, Zenbio, China), and OPG (1:200 dilution, Abcam, UK).

Techniques: Staining