Journal: International Journal of Molecular Sciences

Article Title: Cytoprotective Preconditioning of Osteoblast-Like Cells with N -Acetyl- L -Cysteine for Bone Regeneration in Cell Therapy

doi: 10.3390/ijms20205199

Figure Lengend Snippet: Effects of preconditioning osteoblast-like cells with N -acetyl- L -cysteine (NAC) on bone regeneration in autologous local cell transplantation. ( A ) Scheme indicating outline of the autologous local cell transplantation model. Bone marrow tissue was aspirated from the right femur bone in the first surgery. Bone marrow stromal cells (BMSCs) were cultured on a 10-cm polystyrene dish in an osteogenic differentiation medium for 2 weeks for the purpose of cell growth and differentiation into osteoblast-like cells. At confluence, the cells were labeled with a quantum-dot fluorescent tracer and then preincubated with 5 mM NAC for 3 h. The cells were seeded on a collagen sponge in a NAC-free growth medium. After overnight incubation, the cell-collagen unit was implanted into a critical-sized defect (5 × 5 mm) created on the opposite left femur bone of the donor rat. Representative micro-computed tomography images of the original grayscale cross-section at the long midline of the defect ( B – D ) and overhead view of constructed three-dimensional bone architecture for the defect ( E – G ) 3 weeks after implantation of a collagen sponge alone ( B , E ) or autologous rat osteoblast-like cells with ( D , G ) and without ( C , F ) preincubation with 5 mM NAC for 3 h. Quantitative assessment of three-dimensional bone morphometrical parameters ( H ), bone volume fraction (BV/TV) and trabecular number (Tb. N), thickness (Tb. Th) and separation (Tb. Sp) was performed in the areas of interest in the cortical and bone marrow space regions. The presented data represent the mean ± standard deviation ( n = 4). Asterisks indicate statistically significant differences ( p < 0.05 , Student’s or Welch’s t-test).

Article Snippet: After 14 days (when the cells reached 80% confluence), the cells were labeled with 1 nM fluorescent organic dot cell tracer (Long Term Cell Tracer 500 Cat. # P710G; 101 Bio.com , CA, USA) for 12 h in suspension.

Techniques: Transplantation Assay, Cell Culture, Labeling, Incubation, Micro-CT, Construct, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Cytoprotective Preconditioning of Osteoblast-Like Cells with N -Acetyl- L -Cysteine for Bone Regeneration in Cell Therapy

doi: 10.3390/ijms20205199

Figure Lengend Snippet: Histological features of newly formed bone tissue after autologous local transplantation of osteoblast-like cells with N -acetyl- L -cysteine (NAC). Representative light-microscope images with hematoxylin and eosin ( A , B,E ) or tartrate-resistant acid phosphatase (TRAP) staining ( C , D ) and fluorescent images merged with ( G , H ) and without ( F ) a light-microscope image in the same region on the histological cross section at the long midline of a 5 × 5 mm defect on the rat femur cortical bone 3 weeks after local transplantation of the collagen sponge vehicle used to load autologous rat osteoblast-like cells pretreated with 5 mM NAC for 3 h. Green in the fluorescent images ( F – H ) indicates transplanted osteoblast-like cells prelabeled with a fluorescent cell tracer. Notes: White arrows: location of green fluorescent signals within newly formed bone tissue; white arrowheads: location of green fluorescent signals on the outside of newly formed bone tissue; double arrow: location of TRAP positive spots; black dashed line: a border between the existing and newly formed bone tissue; white dash line: surface of newly formed bone tissue; black dash square in ( A , C ): the regions corresponding to higher magnification images in ( B , D ).

Article Snippet: After 14 days (when the cells reached 80% confluence), the cells were labeled with 1 nM fluorescent organic dot cell tracer (Long Term Cell Tracer 500 Cat. # P710G; 101 Bio.com , CA, USA) for 12 h in suspension.

Techniques: Transplantation Assay, Light Microscopy, Staining