AAGP™ Toxicity in a Keratinocyte Cell Culture

Hussein, 2006

Introduction

It has been hypothesized that AAGP™ may have cell protective properties at concentrations of 1-2mg/ml. With therapeutic applications in mind our aim was to elucidate at what concentration, if any, AAGP™ is toxic to Keratinocyte cell culture.

Method

Keratinocytes were seeded at a concentration of 25,000 cells/well (day -1) and exposed to 0, 1, 2, 5, 10, 20 or 50 mg/ml of AAGP™ (day 0). The cell proliferation of each well (2 wells per concentration) was assessed using Trypan Blue cell counting (2 counts per well) at 24, 48 and 72 hours post exposure. The oxidative activity of the cultures was also measured using the MTT test (2 wells per concentration).

Results

Cell populations exposed to 1-5mg/ml of AAGP™ showed a similar rate of proliferation in comparison to the control groups. Whereas cell populations exposed to 10-20 mg/ml of AAGP™ showed a slowed proliferation and those populations exposed to 50mg/ml exhibited 100% cell death at 72 hours. The MTT test confirmed the toxicity of AAGP™ at a concentration of 50mg/ml. Interestingly, despite cell populations exposed to 1-5mg/ml AAGP™ having a similar cell number in comparison to the control, they appeared to have a reduction in the activity of cell oxidases according to the MTT assay.

Conclusion

These results confirm that AAGP™ is toxic at 50mg/ml and suggest that AAGP™ may be protecting against cell death via an anti oxidative affect at a concentration of 1-5mg/ml.