Cookies on PerkinElmer
PerkinElmer uses cookies to ensure that we give you the best experience possible on our website. This may include cookies from third party websites. If you continue without changing your settings, we will assume that you consent to receive cookies from this website. You can change your cookie settings at any time. To learn more, please review our cookie policy, which includes information on how to manage your cookies.

OsteoSense on the ASK

osteosensefig1.jpg


Overview


Vertebral skeleton development and maintenance depends upon a balance between osteoblast and osteoclast activity. Osteoblasts are cells that are important in the formation of bone, whereas osteoclasts are cells that resorb bone. A change in the balance between these two cell types is characteristic of numerous disease states. These diseases include arthritis, osteoporosis, and cancer metastases. Hydroxyapatite (HA) is a mineral form of calcium apatite and is the major mineral product of osteoblasts. Therefore, HA levels are a good biomarker for osteoblast activity. In addition, abnormal accumulation of HA can be indicative of a disease state. The OsteoSense imaging agents were designed to bind with high affinity to HA both in vitro and in vivo. Since hydroxyapatite (HA) is known to bind pyrophosphonates and phosphonates as well as synthetic bisphosphonates with high affinity, OsteoSense agents were designed as bisphosphonate imaging agents. Specifically, OsteoSense imaging agents can be used to image areas of microcalcifications, bone remodeling and enables imaging of bone growth and resorption.



Products and catalog numbers


ProductCatalog NumberEx/Em wavelength (nm)Molecular weight (g/mol)Validated ExperimentsApplicationsStorage and Stability
OsteoSense 680 EXNEV10020EX668/6871470.5In vivoOncology
Arthritis
Osteoporosis
Technical Data Sheet
OsteoSense 750 EXNEV10053EX749/7701101.1In vivoOncology
Arthritis
Osteoporosis
Technical Data Sheet
OsteoSense 800NEV11105780/8051281In vivoOncology
Arthritis
Osteoporosis
Technical Data Sheet


Using OsteoSense in vivo


OsteoSense agents can be used to measure the effects of therapeutic stimulation of bone growth and can be used to characterize bone remodeling associated with animal models of arthritis.

  • OsteoSense 680 EX and OsteoSense 750 EX are administered via tail vein injection and imaged 24 hours post tail vein injection.
  • OsteoSense 800 is administered via intravenous injection and imaged 24 hours post injection.

For instructions on setting up an in vivo mouse experiment with OsteoSense and imaging on an IVIS or FMT system:

ProductRoute of InjectionMouse Dose (25 g)Rat Dose (250 g)Blood t 1/2Tissue t 1/2Optimal imaging timeOptimal Re-injection Time (complete clearance)Route of Metabolism/ background tissueFMT & IVIS settings
OsteoSense 680EXIV (IP)2 nmol6 nmol5-10 min~ 30 days3-24 hPreimage subtractionBladderFMT 680/700
IVIS 675/720
OsteoSense 750EXIV (IP)4 nmol12 nmol5-10 min7-10 days3-24 hPreimage subtractionBladderFMT 750/770
IVIS 745/800
OsteoSense 800IV (IP)2 nmol6 nmol5-10 min7-10 days3-24 hPreimage subtractionBladderFMT 790/810

In Vivo Imaging Applications:

Imaging Active Bone Remodeling:

osteosensefig2.jpg
Figure 1: OsteoSense 680, OsteoSense 750 or free dyes (0.5 nmoles) were injected intraperitoneally in 1 week-old Balb/c mice. Imaging was performed 24 hours later on a planar Kodak 2000 MM system. The lower chart shows the target to background ratio. This was defined as the fluorescent signal (relative fluorescent units) divided by the adjacent tissue background signal.

Imaging Bone:

osteosensefig3.jpg
Figure 2: X-ray (upper left), reflectance (lower left) and FMT images of a mouse (right). The same mouse was imaged on an FMT system in reflectance and tomographic modes 24 hours after injection with OsteoSense 680. The numbers indicate various bone structures: 1) knee; 2) ankle; 3) pelvis; 4) vertebrae. The fluorescence of the OsteoSense imaging agents on the FMT indicates areas of bone renewal. Using the tomographic mode (right), the FMT allows visualization of deeper pelvic and vertebral structures, which are not visible on reflectance.

Imaging Arthritis:

osteosensefig4.jpg
Figure 3: Arthritis was induced in DBA/1J mice by first injecting of bovine collagen (1 mg/ml) in Complete Freund’s Adjuvant (4 mg/ml M. tuberculosis) subcutaneously at the base of the tail. Then, twenty one days later, the mice were boosted with a subcutaneously injection of collagen (1 mg/ml) in Incomplete Freund’s Adjuvant. The thickness of the paw was measured using a digital Vernier caliper. Imaging was performed in arthritic and healthy (non-induced) mice by injection of OsteoSense 680 (2 nmoles) and imaging on a planar Kodak 2000 MM system 24 hours later. Regions of interests within the paws are highlighted. The target-to-background ratio was defined as the specific fluorescent signal (relative fluorescent units) divided by the adjacent tissue background signal.

Simulated Bone Growth:

osteosensefig5.jpg
Figure 4: Injections of human PTH (1–34) were delivered twice daily subcutaneously onto the calvaria of CF-1 mice for 12 days to induce Calvarial bone growth. The control mice were injected with only the vehicle. Twenty-four hours after injection of OsteoSense 680, imaging was performed. The visible calvaria region shows regions of interest. The target to background ratio was defined as the fluorescent signal (relative fluorescent units) divided by the adjacent tissue background signal.

osteosensefig6.jpg
Figure 5: Immediately following imaging (Figure 5), the mice were sacrificed, calvarias dissected, fixed in buffered formalin and decalcified. The sections were stained with Hematoxylin and Eosin, and the calvaria thickness measured using a Zeiss Axioscope microscope and Axiovision software. From this data, it is clear that OsteoSense 680 can be used to image and quantify stimulated bone growth.


Application notes and posters


  • Poster: Imaging of Cathepsin K activity in rodent models of bone turnover and soft tissue calcification


Citations


Please visit our Citations Library for references using OsteoSense on the IVIS or on the FMT.