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(99m)TC-Methylene diphosphonate uptake at injury site correlates with osteoblast differentiation and mineralization during bone healing in mice.

Zhong ZA, Peck A, Li S, VanOss J, Snider J, Droscha CJ, Chang TA, Williams BO - Bone Res (2015)

Bottom Line: In an ex vivo culture system, calvarial cells were differentiated into osteoblasts with osteogenic medium, pulsed with (99m)Tc-MDP at different time points, and quantitated for (99m)Tc-MDP uptake with a gamma counter.We also found that (99m)Tc-MDP uptake was associated with osteoblast maturation and mineralization in vitro.This study provides direct and biological evidence for (99m)Tc-MDP uptake in osteoblasts during bone healing in vivo and in vitro.

View Article: PubMed Central - PubMed

Affiliation: Center for Skeletal Disease Research, Van Andel Research Institute , Grand Rapids, MI, USA ; Department of Internal Medicine, Center for Musculoskeletal Health, UC Davis Medical Center , Davis, CA, USA.

ABSTRACT
(99m)Tc-Methylene diphosphonate ((99m)Tc-MDP) is widely used in clinical settings to detect bone abnormalities. However, the mechanism of (99m)Tc-MDP uptake in bone is not well elucidated. In this study, we utilized a mouse tibia injury model, single-photon emission computed tomography (gamma scintigraphy or SPECT), ex vivo micro-computed tomography, and histology to monitor (99m)Tc-MDP uptake in injury sites during skeletal healing. In an ex vivo culture system, calvarial cells were differentiated into osteoblasts with osteogenic medium, pulsed with (99m)Tc-MDP at different time points, and quantitated for (99m)Tc-MDP uptake with a gamma counter. We demonstrated that (99m)Tc-MDP uptake in the injury sites corresponded to osteoblast generation in those sites throughout the healing process. The (99m)Tc-MDP uptake within the injury sites peaked on day 7 post-injury, while the injury sites were occupied by mature osteoblasts also starting from day 7. (99m)Tc-MDP uptake started to decrease 14 days post-surgery, when we observed the highest level of bony tissue in the injury sites. We also found that (99m)Tc-MDP uptake was associated with osteoblast maturation and mineralization in vitro. This study provides direct and biological evidence for (99m)Tc-MDP uptake in osteoblasts during bone healing in vivo and in vitro.

No MeSH data available.


Related in: MedlinePlus

Early bone healing and mineralization are noted from 7 days post injury. (a) Transaxial 2-D images of injury sites and sagittal images of injured tibiae at different time points were taken with a micro-CT scanner ex vivo. Arrows indicate the injury sites; (b) Sagittal paraffin sections of the injured sites were stained with pentachrome. Bony tissue is in yellow/golden, and cartilage is bluish.
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fig2: Early bone healing and mineralization are noted from 7 days post injury. (a) Transaxial 2-D images of injury sites and sagittal images of injured tibiae at different time points were taken with a micro-CT scanner ex vivo. Arrows indicate the injury sites; (b) Sagittal paraffin sections of the injured sites were stained with pentachrome. Bony tissue is in yellow/golden, and cartilage is bluish.

Mentions: We monitored the bone healing using ex-vivo micro-CT at each time point, and found that mineralized mass started to form at the periosteum and endosteum adjacent to the injury sites 7 days post-surgery. Significant mineralized mass was present inside the injury sites 14 days post-surgery (Figure 2a). We then decalcified the tibia samples and stained the paraffin sections with pentachrome. Corresponding to the micro-CT data, we found bony tissue began to form in the cavity and cells populated the injury sites 7 days post-surgery (Figure 2b). We observed bony tissue filling the injury sites (Figure 2b), and tartrate-resistant acid phosphatase (TRAP) staining (associated with osteoclast activity) reached the highest level in the injury sites on day 14 post-surgery (Figure S1). These observations suggested that histologic data correlated well with the X-ray photographic data in monitoring bone healing.


(99m)TC-Methylene diphosphonate uptake at injury site correlates with osteoblast differentiation and mineralization during bone healing in mice.

Zhong ZA, Peck A, Li S, VanOss J, Snider J, Droscha CJ, Chang TA, Williams BO - Bone Res (2015)

Early bone healing and mineralization are noted from 7 days post injury. (a) Transaxial 2-D images of injury sites and sagittal images of injured tibiae at different time points were taken with a micro-CT scanner ex vivo. Arrows indicate the injury sites; (b) Sagittal paraffin sections of the injured sites were stained with pentachrome. Bony tissue is in yellow/golden, and cartilage is bluish.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4472149&req=5

fig2: Early bone healing and mineralization are noted from 7 days post injury. (a) Transaxial 2-D images of injury sites and sagittal images of injured tibiae at different time points were taken with a micro-CT scanner ex vivo. Arrows indicate the injury sites; (b) Sagittal paraffin sections of the injured sites were stained with pentachrome. Bony tissue is in yellow/golden, and cartilage is bluish.
Mentions: We monitored the bone healing using ex-vivo micro-CT at each time point, and found that mineralized mass started to form at the periosteum and endosteum adjacent to the injury sites 7 days post-surgery. Significant mineralized mass was present inside the injury sites 14 days post-surgery (Figure 2a). We then decalcified the tibia samples and stained the paraffin sections with pentachrome. Corresponding to the micro-CT data, we found bony tissue began to form in the cavity and cells populated the injury sites 7 days post-surgery (Figure 2b). We observed bony tissue filling the injury sites (Figure 2b), and tartrate-resistant acid phosphatase (TRAP) staining (associated with osteoclast activity) reached the highest level in the injury sites on day 14 post-surgery (Figure S1). These observations suggested that histologic data correlated well with the X-ray photographic data in monitoring bone healing.

Bottom Line: In an ex vivo culture system, calvarial cells were differentiated into osteoblasts with osteogenic medium, pulsed with (99m)Tc-MDP at different time points, and quantitated for (99m)Tc-MDP uptake with a gamma counter.We also found that (99m)Tc-MDP uptake was associated with osteoblast maturation and mineralization in vitro.This study provides direct and biological evidence for (99m)Tc-MDP uptake in osteoblasts during bone healing in vivo and in vitro.

View Article: PubMed Central - PubMed

Affiliation: Center for Skeletal Disease Research, Van Andel Research Institute , Grand Rapids, MI, USA ; Department of Internal Medicine, Center for Musculoskeletal Health, UC Davis Medical Center , Davis, CA, USA.

ABSTRACT
(99m)Tc-Methylene diphosphonate ((99m)Tc-MDP) is widely used in clinical settings to detect bone abnormalities. However, the mechanism of (99m)Tc-MDP uptake in bone is not well elucidated. In this study, we utilized a mouse tibia injury model, single-photon emission computed tomography (gamma scintigraphy or SPECT), ex vivo micro-computed tomography, and histology to monitor (99m)Tc-MDP uptake in injury sites during skeletal healing. In an ex vivo culture system, calvarial cells were differentiated into osteoblasts with osteogenic medium, pulsed with (99m)Tc-MDP at different time points, and quantitated for (99m)Tc-MDP uptake with a gamma counter. We demonstrated that (99m)Tc-MDP uptake in the injury sites corresponded to osteoblast generation in those sites throughout the healing process. The (99m)Tc-MDP uptake within the injury sites peaked on day 7 post-injury, while the injury sites were occupied by mature osteoblasts also starting from day 7. (99m)Tc-MDP uptake started to decrease 14 days post-surgery, when we observed the highest level of bony tissue in the injury sites. We also found that (99m)Tc-MDP uptake was associated with osteoblast maturation and mineralization in vitro. This study provides direct and biological evidence for (99m)Tc-MDP uptake in osteoblasts during bone healing in vivo and in vitro.

No MeSH data available.


Related in: MedlinePlus