environmental regulations. Sharp blades can cause cuts
and should be kept in the appropriate sharps container
and labelled biohazardous after use.
2. In our previous research [7], hind legs with joints from sexually
mature pigs (aged over 54 weeks) were obtained from meat
processing plants, and femoral condyles with intact articular
cartilage were dissected for research. Porcine hind legs were
collected and transported to the laboratory in a plastic cooler.
This research project was performed under animal ethics permission from the animal care committee (University of Alberta
Research Ethics Office).
3. CPA solutions are prepared fresh in PBS solution (use DMEMF12 if cell viability is the interest of investigation). All the CPA
solutions are stirred vigorously at 4
C using an electronic
stirrer for 30 min and then held at the desired temperature
before starting CPA permeation into articular cartilage. We
selected 4, 22, and 37
C to give a range of permeation kinetics
for calculations.
4. This method is for measuring CPA permeation in a pure cartilage disk without the bone base. It is also possible to measure
the CPA permeation in articular cartilage while still attached to
its bone base according to our previous method [6].
5. The 10 mm diameter coring device is used to produce nearly
flat cartilage disks for osmometric measurement. This method
is modified from our previous studies [6, 7, 12] as an advanced
approach to calculate the articular cartilage volume. The cartilage thicknesses are measured at three positions for calculation
of the average height of the cartilage disk. The volume of the
cartilage disk is estimated by the disk bottom area multiplied by
disk height (see Fig. 1). A digital caliper with 0.0005 in. resolution is required to measure the cartilage disk height.
6. There are two range options on the Osmette™ Micro-Osmette
osmometer (Model 5004). The first range “0–2” is for measurement from 0 to 2000 mOsm/kg H 2 O. The second range
“1–3” is for measurement from 1000 to 3000 mOsm/kg
H 2 O, and 1000 needs to be added manually to the reading
value when using the “1–3” range.
7. When the operating head is pressed, the refrigerator module
will start freezing the test tube. The seed wire in the center of
the test sample will induce ice nucleation by physical vibration,
and the thermometer will measure the freezing temperature of
the sample.
8. The measured osmolality of the sample is output on the display
in units of mOsm/kg H 2 O.
314
Kezhou Wu et al.
and should be kept in the appropriate sharps container
and labelled biohazardous after use.
2. In our previous research [7], hind legs with joints from sexually
mature pigs (aged over 54 weeks) were obtained from meat
processing plants, and femoral condyles with intact articular
cartilage were dissected for research. Porcine hind legs were
collected and transported to the laboratory in a plastic cooler.
This research project was performed under animal ethics permission from the animal care committee (University of Alberta
Research Ethics Office).
3. CPA solutions are prepared fresh in PBS solution (use DMEMF12 if cell viability is the interest of investigation). All the CPA
solutions are stirred vigorously at 4
C using an electronic
stirrer for 30 min and then held at the desired temperature
before starting CPA permeation into articular cartilage. We
selected 4, 22, and 37
C to give a range of permeation kinetics
for calculations.
4. This method is for measuring CPA permeation in a pure cartilage disk without the bone base. It is also possible to measure
the CPA permeation in articular cartilage while still attached to
its bone base according to our previous method [6].
5. The 10 mm diameter coring device is used to produce nearly
flat cartilage disks for osmometric measurement. This method
is modified from our previous studies [6, 7, 12] as an advanced
approach to calculate the articular cartilage volume. The cartilage thicknesses are measured at three positions for calculation
of the average height of the cartilage disk. The volume of the
cartilage disk is estimated by the disk bottom area multiplied by
disk height (see Fig. 1). A digital caliper with 0.0005 in. resolution is required to measure the cartilage disk height.
6. There are two range options on the Osmette™ Micro-Osmette
osmometer (Model 5004). The first range “0–2” is for measurement from 0 to 2000 mOsm/kg H 2 O. The second range
“1–3” is for measurement from 1000 to 3000 mOsm/kg
H 2 O, and 1000 needs to be added manually to the reading
value when using the “1–3” range.
7. When the operating head is pressed, the refrigerator module
will start freezing the test tube. The seed wire in the center of
the test sample will induce ice nucleation by physical vibration,
and the thermometer will measure the freezing temperature of
the sample.
8. The measured osmolality of the sample is output on the display
in units of mOsm/kg H 2 O.
314
Kezhou Wu et al.