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Table 1 Production steps of insoluble bovine collagenous bone matrices ICBM

From: Release kinetics of the model protein FITC-BSA from different polymer-coated bovine bone substitutes

Step 1Coarse degreasing of femur bone marrow with a steam jet
Step 2Thorough degreasing of femur bone marrow by application of chloroform:methanol (3:1) solution for three consecutive times at 24 h, 72 h and 24 h intervals.
Step 3Washing with distilled water, bleaching by application of hydrogen peroxide [H2O2] (3% for 15 min), washing with distilled water.
Step 4Demineralization by application of 0.5 M (M) hydrogen chloride [HCL] for three consecutive times at 1.5 h intervals.
Step 5Washing with distilled water, sawing out the cuboid femur slices, application of 0.5 M HCL for 1 h, washing with distilled water, inactivation of osteoinductive matrix proteins by application of 4 M guanidine-HCL/Tris*-HCL (pH value 7.0) for 24 h at 4 °C.
Step 6Application of 50 mM Tris-HCL/150 mM sodium chloride [NaCl] (pH value 7.0), washing with distilled water for 30 min, temporary freezing at − 80 °C for 72 h, lyophilization at − 4 °C and 0.05 mbar for 24 h, storage at 4 °C until gamma sterilization and final sterile storage in 50 ml containers at room temperature (RT).
Step 7Assorting of ICBM with medium cancellous bone density for use in this study.
  1. * tris (hydroxymethyl)aminomethane