Physical characterization of succinylated type I collagen by Raman spectra and MALDI-TOF/MS and in vitro evaluation for biomedical applications

In this study, we report on physical and in vitro biological characterization of succinylated collagen (SC). SC was prepared by succinylation of type I bovine tendon collagen. SC swells and dissolves in physiological pH buffers (pH 7.4) Biocompatibility of SC to collagen for fibroblasts was comparable but L6 myoblasts showed pronounced proliferation and differentiation with SC. Using the MALDI-TOF/MS technique, SC was found with increased molecular mass by 16,359 Da per molecule which corresponds to about 54 succinyl groups covalently linked to the collagen strand. Raman spectroscopy revealed the retention of triple helical structure conformation in the presence of linked succinyl groups. New peaks near 1737, 1675 and 1420 cm−1 and decreased intensities near 2440 and 488 cm−1 provides the most convenient marker bands for succinylation of collagen. The intense band regions near 2856–2934, 2724, and 1445 cm−1 also confirms the existence of succinyl groups.

► Succinylated collagen (SC) prepared by succinylation of type I bovine tendon collagen.
► Isolation and culture of rat cardiac fibroblasts.
► Biocompatibility assay for SC and collagen by using NIH 3T3fibroblasts, rat cardiac fibroblasts and L6 myoblasts.
► Raman spectroscopy and MALDI-TOF/MS for SC and collagen.
► Cytoproliferative effects for collagen matrix.