Effect of Different Methods of Trypsinization on Cell Viability and Clinical Outcome in Vitiligo Patients Undergoing Noncultured Epidermal Cellular Suspension.

BACKGROUND: Noncultured Epidermal Cell Suspension (NCECS) is a surgical modality used in treating stable vitiligo. Trypsinization of the epidermis may be done either at 4°C overnight (cold) or at 37°C for 30 to 50 minutes (warm). Recently, trypsinization was done at room temperature (25°C) in an in vitro trial. OBJECTIVE: To compare different trypsinization techniques in NCECS regarding cell viability and clinical outcome. METHODS: This comparative multicenter study was conducted on 20 patients with stable nonsegmental vitiligo. In each patient, 3, nonacral vitiligo lesions were randomly assigned for treatment by NCECS prepared by warm, room temperature, and cold trypsinization techniques, respectively. A perilesional biopsy was taken from each of the 3 treated lesions as an objective measure of disease stability. After transplantation, all patients received narrow-band ultraviolet B twice weekly for 6 months. Cell viability was assessed in each technique, as well as clinical outcome in all treated lesions. RESULTS: Warm and room temperature trypsinization techniques were comparable with each other. Both were significantly better than the cold technique regarding viability and repigmentation. CONCLUSION: Room temperature trypsinization can be used as a convenient substitute to warm trypsinization. Cold trypsinization is not recommended because of its poor results and poor patient satisfaction.

Antiangiogenic effect of methotrexate and PUVA on psoriasis.

Vascular endothelial growth factor (VEGF) is important factor for angiogenesis in psoriasis. Methotrexate and psoralen and ultraviolet light A (PUVA) mainly target the T cell-mediated immunopathology of psoriasis. Our work aimed at estimating VEGF mRNA in psoriatic patients and investigating whether the standard therapeutic modalities (methotrexate and PUVA) exert their antiangiogenic activity through altering VEGF levels. Twenty-four chronic plaque psoriasis patients were enrolled. Patients were divided into two groups (12 patients each); group A received intramuscular methotrexate and group B was treated by PUVA three times/week in a PUVA 1000 cabin for 10 weeks each. Twelve healthy volunteers served as controls. A skin biopsy was taken from lesional skin before and after treatment for RT-PCR detection of VEGF mRNA. Capillary perfusion scanning using LASER Doppler perfusion imaging was performed on the same psoriatic plaque before and after treatment and was also done for the controls. Following both methotrexate and PUVA, a significant reduction in the amount of VEGF mRNA (P < 0.001 and P = 0.002, respectively) and capillary perfusion (P = 0.002) occurred. These reductions were significantly higher in the methotrexate group (P < 0.001 and  P = 0.001, respectively) than in the PUVA group. The percentage of clinical improvement in the examined psoriatic plaque was significantly positively correlated with the percentage of reduction in the amount of VEGF mRNA (r = 0.850, P < 0.001) and the percentage of reduction in the capillary perfusion (r = 0.684, P < 0.001). Both modalities may exert an antiangiogenic effect. Methotrexate appears to have possibly a more potent antiangiogenic effect than PUVA.