Figure 6.
In vivo pharmacological treatment accompanying transplantation process rescues bone biology in mice. (A) Experimental design for HSCT into lethally irradiated 3-month-old wild-type mice (time point at the end of experiment: 13 months, 13R). Age-matched control group (13A) without HSCT. In vivo injection of CASIN (IP, blue filled symbols) at days 5, 6, 7, and 8 after HSCT (13R). Vector injection (phosphate-buffered saline and 15% ethanol, gray-filled symbols) at the same time points. Analysis of the BM niche of 13-month-old mice (13R with or without CASIN and 13A with or without CASIN) and Y control group (3 months, no HSCT; no CASIN and/or vehicle treatment). CT imaging of young and 13-month-old mice (13R + CASIN, 13R + vehicle, and 13A (13A no HSCT; no CASIN and/or vehicle treatment). (B) Graph shows the total cell number out of 4 long BM flushed bones after collagenase digest with CASIN or vehicle treatment (left graph) and relative numbers of immature MSPCs (CD45/Ter119/CD31− Sca-1+ Alcam−/low, middle left) and OBCs (CD45/Ter119/CD31− Sca-1− Alcam+, middle right). (C) Graph shows ROS (reactive oxygen levels) staining measured with FACS analysis in cultured MSPC (p4) with representative FACS plot (right). 13R + vehicle, black; 13R + CASIN, red. (D) Graph shows tetramethylrhodamine methyl ester (mitochondrial membrane potential) staining measured with FACS analysis in cultured MSPC (p4) with representative FACS plot (right). 13R + vehicle, black; 13R + CASIN, red. (E) Micro-CT image of 1 dissected long bone (femur) per mouse (all males). Percentage of BV relative to total volume (TV, graph left) measured with ImageJ software and representative micro-CT images of 13R + CASIN (right). (F) Trabecular thickness analyzed with micro-CT imaging with representative micro-CT image of trabecular structures in mouse femur of 13R + CASIN (right). (G) Cortical thickness analyzed with micro-CT imaging. The analysis represents 2-3 independent experiments. The micro-CT imaging was performed once. ∗P < .05 (Kruskal-Wallis test: panels B-G). Data are represented as mean ± SD. Cs.Th, cortical thickness; PB, peripheral blood; Tb.Th, trabecular thickness.

In vivo pharmacological treatment accompanying transplantation process rescues bone biology in mice. (A) Experimental design for HSCT into lethally irradiated 3-month-old wild-type mice (time point at the end of experiment: 13 months, 13R). Age-matched control group (13A) without HSCT. In vivo injection of CASIN (IP, blue filled symbols) at days 5, 6, 7, and 8 after HSCT (13R). Vector injection (phosphate-buffered saline and 15% ethanol, gray-filled symbols) at the same time points. Analysis of the BM niche of 13-month-old mice (13R with or without CASIN and 13A with or without CASIN) and Y control group (3 months, no HSCT; no CASIN and/or vehicle treatment). CT imaging of young and 13-month-old mice (13R + CASIN, 13R + vehicle, and 13A (13A no HSCT; no CASIN and/or vehicle treatment). (B) Graph shows the total cell number out of 4 long BM flushed bones after collagenase digest with CASIN or vehicle treatment (left graph) and relative numbers of immature MSPCs (CD45/Ter119/CD31− Sca-1+ Alcam−/low, middle left) and OBCs (CD45/Ter119/CD31− Sca-1− Alcam+, middle right). (C) Graph shows ROS (reactive oxygen levels) staining measured with FACS analysis in cultured MSPC (p4) with representative FACS plot (right). 13R + vehicle, black; 13R + CASIN, red. (D) Graph shows tetramethylrhodamine methyl ester (mitochondrial membrane potential) staining measured with FACS analysis in cultured MSPC (p4) with representative FACS plot (right). 13R + vehicle, black; 13R + CASIN, red. (E) Micro-CT image of 1 dissected long bone (femur) per mouse (all males). Percentage of BV relative to total volume (TV, graph left) measured with ImageJ software and representative micro-CT images of 13R + CASIN (right). (F) Trabecular thickness analyzed with micro-CT imaging with representative micro-CT image of trabecular structures in mouse femur of 13R + CASIN (right). (G) Cortical thickness analyzed with micro-CT imaging. The analysis represents 2-3 independent experiments. The micro-CT imaging was performed once. ∗P < .05 (Kruskal-Wallis test: panels B-G). Data are represented as mean ± SD. Cs.Th, cortical thickness; PB, peripheral blood; Tb.Th, trabecular thickness.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal