Journal: bioRxiv
Article Title: A Tough Biointerface in Human Knee Empowered by Dynamic Phase-transforming Minerals in Collagenous Matrix
doi: 10.1101/2024.08.03.606023
Figure Lengend Snippet: ( A ) Schematic illustration of the experimental procedure of proteomic analysis. ( B ) Classification of interface-enriched proteins based on functions described in Genecards Database. ( C ) Volcano plots of protein expression differences between interface and bone, and interface and root. Proteins with statistically significant differences in expression were colored in orange (significantly higher in interface) and blue (significantly lower in interface) ( p adjusted value < 0.05, fold change > 2). ( D ) Immunofluorescent staining of MGP at the S-H interface. ( E ) MGP protein expression profile across the S-H interface quantified using fluorescent signal intensity which increased in the mineralized tissue region and S-H interface (denoted by dashed line). ( F ) Functional verification of MGP’s role as a mineralization inhibitor through in vitro mineralization using nano-ACP. TEM images showed the increase in HAP crystals in the non-treated group was less than in the MGP-treated group on day 1 and day 3. Inlets revealed diffraction patterns of associated areas. ( G ) Quantification of HAP crystal density in TEM snapshots (n=5) of MGP-treated and non-treated groups. Statistical significance is illustrated as: *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001. Abbreviations: LC-MS/MS, liquid chromatography−tandem mass spectrometry; MGP, matrix Gla protein.
Article Snippet: 200-mesh golden grids with a carbon support film were added with 3 μL recombinant matrix gla protein (MGP) (100 μg/mL, CUSABIO, China) diluted with deionized water and the control group was added with 3 μL deionized water, and both were incubated for 20 minutes at room temperature.
Techniques: Expressing, Staining, Functional Assay, In Vitro, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Mass Spectrometry