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. Author manuscript; available in PMC: 2016 Aug 11.

Published in final edited form as: Wound Repair Regen. 2015 Nov 4;23(6):842–854. doi: 10.1111/wrr.12365

A) Twenty BALB/c mice were randomized to four treatment groups. Two splinted full thickness wounds were made on the backs of each mouse, and were treated with two 8mm diameter discs of Telfa® pads soaked with 60μl of either PBS (Control), 50mM D-tryptophan, 50mM L-tryptophan, or a 50:50 combination of D- and L-tryptophan (50mM total tryptophan concentration). Treatments were applied on day 0 and reapplied on days 3 and 6. Images of the wounds were taken on days 0 (baseline), 3, 6, and 9 with a Nikon D300 camera. Image J was used to calculate the wound areas, which were normalized to the baseline (day 0) values. No significant differences in remaining wound area were measured between any treatment and the control, a p-value of below 0.05 was considered significant. B) A one hour exposure of the immortalized HaCaT cell line to D-tryptophan at 37°C exhibited no direct cytotoxicity. Cell viability was measured using calcein-AM and measuring resulting fluorescence at EM:485nm/EX:528nm. C) No loss of cellular membrane integrity was observed in immortalized human keratinocytes (STINKs) with D-/L-tryptophan concentrations up to 10mM over 24 and 48 hour incubations at 37°C using the CellTox^™ Green Cytotoxicity Assay. D) Altered or reduced the cellular metabolism of the STINKs cell line was observed over 24 and 48 hour incubations with increasing D-/L-tryptophan concentrations as assessed by the RealTime-Glo MT Cell Viability Assay.

A) Twenty BALB/c mice were randomized to four treatment groups. Two splinted full thickness wounds were made on the backs of each mouse, and were treated with two 8mm diameter discs of Telfa® pads soaked with 60μl of either PBS (Control), 50mM D-tryptophan, 50mM L-tryptophan, or a 50:50 combination of D- and L-tryptophan (50mM total tryptophan concentration). Treatments were applied on day 0 and reapplied on days 3 and 6. Images of the wounds were taken on days 0 (baseline), 3, 6, and 9 with a Nikon D300 camera. Image J was used to calculate the wound areas, which were normalized to the baseline (day 0) values. No significant differences in remaining wound area were measured between any treatment and the control, a p-value of below 0.05 was considered significant. B) A one hour exposure of the immortalized HaCaT cell line to D-tryptophan at 37°C exhibited no direct cytotoxicity. Cell viability was measured using calcein-AM and measuring resulting fluorescence at EM:485nm/EX:528nm. C) No loss of cellular membrane integrity was observed in immortalized human keratinocytes (STINKs) with D-/L-tryptophan concentrations up to 10mM over 24 and 48 hour incubations at 37°C using the CellTox^™ Green Cytotoxicity Assay. D) Altered or reduced the cellular metabolism of the STINKs cell line was observed over 24 and 48 hour incubations with increasing D-/L-tryptophan concentrations as assessed by the RealTime-Glo MT Cell Viability Assay.