Journal: bioRxiv

Article Title: In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale

doi: 10.1101/2023.10.10.560919

Figure Lengend Snippet: Optimization of different conditions for mineralizing microgels. (A) Microgels were mineralized with different conditions of photopolymerization time and media composition. (B) and (C) represent the mineralized matrix ratio and the crystallinity index of microgels cultivated in DMEM low and high glucose, as well as α-MEM media. (D-F) represent the mineralized matrix ratio, the crystallinity index, and the elastic moduli in kPa of microgels (GelMA 10%) that were exposed to 25, 50, or 100 seconds in the digital light printing (DLP) process. Statistical differences are represented by * (p<0.05), ** (p<0.01), and *** (p<0.001) after one-way or two-way ANOVA and a post-Tukey’s correction test.

Article Snippet: The number of cells, number of dendrites per cell and the length of dendrites per osteocytes in bioprinted microgels were quantified using Imaris 8.0.1 (Oxford Instruments, Tubney Woods, AB, UK) and ImageJ/FIJI [ ] after 3 days of mineralization.

Techniques:

Journal: bioRxiv

Article Title: In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale

doi: 10.1101/2023.10.10.560919

Figure Lengend Snippet: Printing and mineralization of microgels shapes. (A) Schematic of microgel preparation using DLP printer and mineralization steps. (B – H) represent the printed shapes, including triangle, square, circle, heart, stars, and the OHSU logo. (C, F, J) show the printed microgels in phase contrast. (D, G, K) show microgels imaged in green fluorescence. (E,H, L) show the printed constructs stained with alizarin red.

Article Snippet: The number of cells, number of dendrites per cell and the length of dendrites per osteocytes in bioprinted microgels were quantified using Imaris 8.0.1 (Oxford Instruments, Tubney Woods, AB, UK) and ImageJ/FIJI [ ] after 3 days of mineralization.

Techniques: Fluorescence, Construct, Staining

Journal: bioRxiv

Article Title: In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale

doi: 10.1101/2023.10.10.560919

Figure Lengend Snippet: Microgel characterization. The images show the morphology of mineralized and non-mineralized microgels in 4 and 10x of magnification by light microscopy. B and C show the differences in morphology and extrafibrillar mineralization between mineralized and non-mineralized microgels from top view and from a cross-section view by scanning electron (B) and transmission electron microscopy (C).

Article Snippet: The number of cells, number of dendrites per cell and the length of dendrites per osteocytes in bioprinted microgels were quantified using Imaris 8.0.1 (Oxford Instruments, Tubney Woods, AB, UK) and ImageJ/FIJI [ ] after 3 days of mineralization.

Techniques: Light Microscopy, Transmission Assay, Electron Microscopy

Journal: bioRxiv

Article Title: In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale

doi: 10.1101/2023.10.10.560919

Figure Lengend Snippet: Characterization of non-mineralized and mineralized microgels. Mineralized microgels stained with Alizarin red (A) and von Kossa (B) show dark solid areas compared to the non-mineralized group after 3 days. By the energy diffraction x-ray analysis (EDAX), (C) the presence of Ca and P electron peaks were just observed in mineralized microgels. (D). FTIR analysis shows the absorbance of different organic (amides I-III) and inorganic (phosphate) chemical groups only on mineralized samples with the wavenumber expressed in cm-1. The mineral matrix ratio (E), the crystallinity index (F) and the elastic modulus confirm higher stiffness (represented in kPa) and solid composition of mineralized microgels compared to the non-mineralized samples. Statistical differences are represented by ** p<0.01, and **** p<0.0001 after the One-way ANOVA test post-Turkey’s corrections.

Article Snippet: The number of cells, number of dendrites per cell and the length of dendrites per osteocytes in bioprinted microgels were quantified using Imaris 8.0.1 (Oxford Instruments, Tubney Woods, AB, UK) and ImageJ/FIJI [ ] after 3 days of mineralization.

Techniques: Staining

Journal: bioRxiv

Article Title: In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale

doi: 10.1101/2023.10.10.560919

Figure Lengend Snippet: Osteocyte morphology and functionality in mineralized and non-mineralized microgels. A-B and E-F represent the confocal images of non-mineralized (A-B) and mineralized (E-F) samples. Microgels stained with actin, DAPI, and SOST revels higher cell number and SOST intensity (normalized fluorescence per cell) in mineralized microgels (A-H, K-L). Also, the dendrite length in μm (I) and the number of dendrites per cell (I-J) were not significantly different between mineralized and non-mineralized samples. Statistical differences are represented by **** p<0.0001 after the One-way ANOVA test post-Turkey’s corrections.

Article Snippet: The number of cells, number of dendrites per cell and the length of dendrites per osteocytes in bioprinted microgels were quantified using Imaris 8.0.1 (Oxford Instruments, Tubney Woods, AB, UK) and ImageJ/FIJI [ ] after 3 days of mineralization.

Techniques: Staining, Fluorescence

Journal: PLoS ONE

Article Title: Domain of Dentine Sialoprotein Mediates Proliferation and Differentiation of Human Periodontal Ligament Stem Cells

doi: 10.1371/journal.pone.0081655

Figure Lengend Snippet: A . Coomassie-stained SDS-PAGE gel for analysis of expression of the recombinant COOH-terminal dentin sialoprotein (rC-DSP) protein. The sample in DL21 cells was grown 3 h with 1 mM IPTG induction after reaching O.D.600 of 0.6. The rC-DSP was purified by the column described in the materials and methods. B . Expression of purified rC-DSP was confirmed by Western blotting analysis using goat polyclonal anti-DSP antibody (Santa Cruz Biotechnology Inc.). D - E . Collagen type I and rC-DSP proteins were coated on glass slides for overnight at 37°C. For scanning electron microscopy, the samples were washed and dried as well as treated with sodium deoxycholate. Then the samples were observed under scanning electron microscope (SEM). Fibril matrix was visible on the collagen type I coated slide ( D ) and rC-DSP coated slide ( E ), but no control slide ( C ). Also, there were different morphologies between rC-DSP and collagen type I.

Article Snippet: Rabbit polyclonal anti-human DMP1 (a gift from Dr. Larry Fisher, National Institute of Health, Bethesda, MD), rabbit polyclonal anti-human DLX3 and rabbit polyclonal anti-human OSX (Abcam), rabbit polyclonal anti-human osteocalcin (OC) antibody, goat polyclonal anti-human osteoprotegerin (OPG) antibody and goat polyclonal anti-human RUNX2 antibody (Santa Cruz Biotechnology, Inc.) were used as primary antibodies.

Techniques: Staining, SDS Page, Expressing, Recombinant, Purification, Western Blot, Electron Microscopy, Microscopy, Control