Ground type-I collagen—a focused study on its fibrillogenesis behavior and bioactivity in vitro

다국어 초록 (Multilingual Abstract)

Take bovine tendon type-I collagen as the raw material, we pretreated it by freezing and then grinding the macromolecule at a low temperature (4–10 °C). Effects of the grinding process on the molecular structure, water solubility, fibrillogenesis behavior in vitro and biological activity of collagen were systematically prospected. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) results revealed that the collagen had typical structural characteristics of type-I collagen; moreover, grinding treatment gave rise to partially degraded collagen molecules which took on continuous bands with lower molecular weights different from α1 and α2. More strikingly, the degradation degree positively corresponded to the grinding duration. Circular dichroism (CD) analysis manifested that with extended grinding, intensities of both the positive and negative peaks for collagen samples shrunk. Interestingly, grinding also contributed to improved water solubility of collagen. The water solubility of the sample ground for 20 min (COL20) was 1.91 mg/mL, which is about 6 times higher than that of the natural collagen sample (0.28 mg/mL). Amid the study concerning the fibrillogenesis behavior, turbidity and rheological data collaboratively demonstrated that with prolonged grinding time, the assembly rate and degree of the sample plunged. Assembled gel from the ground collagen with finer fibrils was also observed in scanning electron microscope (SEM), which further embraced the conclusion aforementioned in fibrillogenesis kinetics. Cell experiments corroborated that the grinding process can be harnessed to manipulate the cell proliferation promoting ability of collagen. Simultaneously, further dissection of COL20 with the best biological activity clarified that the water-soluble part and water-insoluble part of COL20 had distinct aptitudes as to promoting cell proliferation as well as inhibiting the growth of liver cancer cells.