An in vitro cytotoxicity study of aldehyde-treated pig dermal collagen

The cytotoxicity of aldehyde-treated collagen was assayed by measuring 3H-thymidine incorporation in adult human skin fibroblasts grown in tissue culture for 1 or 3 days in the presence of pig dermal collagen cross-linked with formaldehyde or glutaraldehyde. A comparison was also made with collagen preparations washed for 2 weeks either at 15 degrees throughout or partly at 15 degrees and partly at 37 degrees. Collagen treated with both formaldehyde and glutaraldehyde proved increasingly toxic with increase in the concentrations of aldehyde used. While the maximum toxic effect was observed after 1 day culture in formaldehyde-treated collagen, with thymidine uptake ranging from 4-48% of control values with 5-0.1% formaldehyde and a 15 degrees wash, the toxic effect of glutaraldehyde treatment increased with longer exposure and at 3 days thymidine uptake ranged from 3-40% of control values with 0.05-0.001% glutaraldehyde and washing at 15 degrees. Washing partly at 37 degrees significantly reduced toxicity, the differences in thymidine uptake as compared with washing at 15 degrees alone ranging from 34-50% with 1 and 0.3% formaldehyde respectively in 1 day cultures and from 14-37% with 0.02 and 0.005% glutaraldehyde in 3 day cultures. While fibroblasts actively grew and migrated when seeded on non-cross-linked collagen, only limited cell survival occurred on aldehyde-treated collagen.