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Bone matrix integrity in response to HHP treatment. Human trabecular bone was treated with HHP at 250, 350, and 600 MPa at 10°C for 20 min using 0.9% saline as the pressure-transmitting medium before post-treatment incubation of bone cylinders was conducted at 4°C for 1 h. <t>Multiphoton</t> microscopy images (a–j), showing tissue autofluorescence at 525/50 nm (green; organic matrix components)) and second harmonic generation (SHG) signal at 405/20 nm (blue; fibrillar collagen), using an excitation wavelength of 810 nm. Here, treated bone cylinders (c–h) are compared to untreated bone (a and b) and bone that was autoclaved (i and j) 121°C for 20 min). Scale bar: 100 µm. The collagen I Western blot (k) is based on a native PAGE (1000 ng of protein per band) with the corresponding quantification (l) that was performed using Image Lab quantification of respective band intensities. Mechanical characterization of human trabecular bone cylinders ( n = 9) regarding compressive modulus (m) and strength (n) via a tactile extensometer. Compressive modulus and strength are displayed as means with standard deviation and single values, represented as dots colored according to corresponding bone density values. Statistical analysis of M and N: Kruskal-Wallis test with Dunn’s multiple comparisons test: p > 0.05.
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Bone matrix integrity in response to HHP treatment. Human trabecular bone was treated with HHP at 250, 350, and 600 MPa at 10°C for 20 min using 0.9% saline as the pressure-transmitting medium before post-treatment incubation of bone cylinders was conducted at 4°C for 1 h. <t>Multiphoton</t> microscopy images (a–j), showing tissue autofluorescence at 525/50 nm (green; organic matrix components)) and second harmonic generation (SHG) signal at 405/20 nm (blue; fibrillar collagen), using an excitation wavelength of 810 nm. Here, treated bone cylinders (c–h) are compared to untreated bone (a and b) and bone that was autoclaved (i and j) 121°C for 20 min). Scale bar: 100 µm. The collagen I Western blot (k) is based on a native PAGE (1000 ng of protein per band) with the corresponding quantification (l) that was performed using Image Lab quantification of respective band intensities. Mechanical characterization of human trabecular bone cylinders ( n = 9) regarding compressive modulus (m) and strength (n) via a tactile extensometer. Compressive modulus and strength are displayed as means with standard deviation and single values, represented as dots colored according to corresponding bone density values. Statistical analysis of M and N: Kruskal-Wallis test with Dunn’s multiple comparisons test: p > 0.05.
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Bone matrix integrity in response to HHP treatment. Human trabecular bone was treated with HHP at 250, 350, and 600 MPa at 10°C for 20 min using 0.9% saline as the pressure-transmitting medium before post-treatment incubation of bone cylinders was conducted at 4°C for 1 h. Multiphoton microscopy images (a–j), showing tissue autofluorescence at 525/50 nm (green; organic matrix components)) and second harmonic generation (SHG) signal at 405/20 nm (blue; fibrillar collagen), using an excitation wavelength of 810 nm. Here, treated bone cylinders (c–h) are compared to untreated bone (a and b) and bone that was autoclaved (i and j) 121°C for 20 min). Scale bar: 100 µm. The collagen I Western blot (k) is based on a native PAGE (1000 ng of protein per band) with the corresponding quantification (l) that was performed using Image Lab quantification of respective band intensities. Mechanical characterization of human trabecular bone cylinders ( n = 9) regarding compressive modulus (m) and strength (n) via a tactile extensometer. Compressive modulus and strength are displayed as means with standard deviation and single values, represented as dots colored according to corresponding bone density values. Statistical analysis of M and N: Kruskal-Wallis test with Dunn’s multiple comparisons test: p > 0.05.

Journal: Journal of Tissue Engineering

Article Title: Comprehensive characterization of cell and tissue responses toward high hydrostatic pressure treatment: Molecular feedback and structural integrity in bone graft processing

doi: 10.1177/20417314251337193

Figure Lengend Snippet: Bone matrix integrity in response to HHP treatment. Human trabecular bone was treated with HHP at 250, 350, and 600 MPa at 10°C for 20 min using 0.9% saline as the pressure-transmitting medium before post-treatment incubation of bone cylinders was conducted at 4°C for 1 h. Multiphoton microscopy images (a–j), showing tissue autofluorescence at 525/50 nm (green; organic matrix components)) and second harmonic generation (SHG) signal at 405/20 nm (blue; fibrillar collagen), using an excitation wavelength of 810 nm. Here, treated bone cylinders (c–h) are compared to untreated bone (a and b) and bone that was autoclaved (i and j) 121°C for 20 min). Scale bar: 100 µm. The collagen I Western blot (k) is based on a native PAGE (1000 ng of protein per band) with the corresponding quantification (l) that was performed using Image Lab quantification of respective band intensities. Mechanical characterization of human trabecular bone cylinders ( n = 9) regarding compressive modulus (m) and strength (n) via a tactile extensometer. Compressive modulus and strength are displayed as means with standard deviation and single values, represented as dots colored according to corresponding bone density values. Statistical analysis of M and N: Kruskal-Wallis test with Dunn’s multiple comparisons test: p > 0.05.

Article Snippet: The multiphoton images were taken utilizing an ultra-fast laser-scanning multiphoton microscope (TriMScope II, LaVision BioTec, Bielefeld, Germany) in combination with a mode-locked femtosecond-pulsed Ti:Sa laser (Chameleon Vision II, Coherent, Santa Clara, CA, USA).

Techniques: Saline, Incubation, Microscopy, Western Blot, Clear Native PAGE, Standard Deviation