Immunological evaluation into the bone allograft and selection of experimental animal model on im-munogenicity / 中华创伤骨科杂志

ABSTRACT

Objective To explore the effect of immunogenicity of freeze-dried bone allograft on different in vitro experimental models. Methods The lymphocytes were obtained respectively from 10 healthy young human volunteers, 10 Balb/c and 10 C57 mice and 10 New Zealand rabbits. The experiment was carried out in 6 groups positive control group (PHA/ConA+lymphocyte), negative control group (Hydroxyapatite powder + lymphocyte), allogeneic bone group A (Freeze-dried bone powder 2. 0 g/L + lym-phocyte), allogeneic bone group B (Freeze-dried bone powder 1.0 g/L + lymphocyte), allogeneic bone group C (Freeze-dried bone powder 0.5 g/L + lymphocyte), and negative control group (culture solution + lym-phocyte). Lymphocyte transformation test (Alamarblue) was conducted to culture the 6 kinds of experimental materials in vitro. After 72 hours, samples were scanned with ELISA muhiscan at wave lengths 570 nm and 600 nm to fetal the light absorption value. Pearson analyses were performed 10 determine the relationships a-mong the 3 animals and 1 human groups and find out which animal would be highly correlated to human. Results In the human and Balb/c mice lymphocyte transformation tests, there was no significant difference (P > 0.05) between allogeneic bone groups A, B, C and negative control group (HA) ; but there was sig-nificant difference (P < 0.001) between allogeneic bone groups A, B, C and positive control group (PHA/ConA); there was no significant difference between the 3 allogeneic bone groups (P > 0.05). There was no significant difference among the 6 groups of C57 mice and New Zealand rabbits (P > 0.05). The coefficient r between Balb/c mice and human groups was 0.959, P = 0.003, showing a highly positive correlation. The coefficient r between C57 mice and human groups was 0.527, P = 0.283, while the coefficient r between New Zealand rabbits and human groups was 0.866, P =0.026. Conclusions The immunogenicity of freeze-dried bone powder in this experiment may not be sufficient enough to induce significanrt immunologic response. Balb/c mice may be preferable for immunogenicity related experiments.