Type-I Pepsin-Solubilized Collagen From Bohadschia Bivittata Enhanced hPDLFs Cells Viability

Objectives:
Collagen has been found to have promising role in tissue healing and regeneration. In previous study, we isolated 65% of pepsin-solubilized collagen (PSC) from the body wall of sea cucumber species, Bohadschia bivittata, classified as type-I collagen. The purpose of this study was to examine the cytotoxic effects of non-γ or γ-irradiated PSC on hPDL fibroblasts (hPDLFs).

Methods:
PSC was sterilized by γ-rays at 25 kGy from ⁶⁰Co, and two groups (γ-irradiated PSC or non-γ-irradiated PSC) were made. For cytotoxicity experiment, PSC was added in α-MEM and incubated for 5 days at 37°C. On day 4, the hPDLF (ScienCell^TM, USA) cells were cultured in 96-well plates at 37°C, and 5% CO₂ for 24 hours, and then consequently the PSC extract was added into the wells at seven serial dilutions for 72 hours, and finally MTT assay was used to measure cell viability. Independent t-test was used to determine the significance of the difference between means of non-γ and γ-irradiated PSC extracts at different concentrations on hPDLF cell line. The level of significance (p value) was set at 0.05.
Results:
A significant increase of the cell viability was detected (maximum 3 to 4-times effects at 25 mg/ml), indicating that PSC is non-cytotoxic. However, cell viability values of hPDLFs by γ-irradiated PSC were lower than non-γ-irradiated PSC at from 6.25 to 50 mg/ml (p < 0.05).
Conclusions:
Results indicated that the PSC extracts from B. bivittata did not induce cytotoxicity in hPDLF, and irradiation at 25kGy may have harmful affects on its physiochemical properties. This exploratory study suggests that collagen from B. bivittata could potentially replace mammalian collagen in healing and regenerative procedures.