Simplified cellular grafting for treatment of vitiligo and piebaldism: the "6-well plate" technique.

BACKGROUND: Vitiligo is a disfiguring depigmenting dermatosis that affects approximately 0.5% to 1% of the general population regardless of race and sex. In patients with stable vitiligo who fail conventional therapies, surgical transplant offers a viable alternative. Noncultured cellular grafting offers the advantage of repigmenting vitiligo 5 to 10 times the size of the donor skin and can be completed on the same day on an outpatient basis. In recent years, ways to simplify this procedure have been explored, including the use of commercially available kits. OBJECTIVES: To simplify the extraction of epidermal cells from donor skin using a 6-well plate and to evaluate the clinical efficacy of this simplified technique in repigmenting stable vitiligo and piebaldism. METHODS: Four patients with focal or segmental vitiligo and one with piebaldism were treated using the simplified noncultured cellular grafting protocol. Percentage of repigmentation 6 months after grafting was objectively measured using digital contour mapping. RESULTS: Patients with stable segmental or focal vitiligo achieved 65% to 92% repigmentation 6 months after grafting; the treated sites involved face or limbs. The patient with piebaldism achieved 86% repigmentation. One year after grafting, the extent of repigmentation remained for all patients. CONCLUSION: This set-up is simple and inexpensive; it reduces cell preparation time, amount of reagents used, and costs, and obviates the need of a laboratory for extraction of epidermal cells.

Influence of cryopreservation on human periodontal ligament cells in vitro.

Cryopreservation of teeth before autotransplantation may create new possibilities in dentistry. The purpose of this study was to examine the effect of a standardised cryopreservation procedure on human periodontal ligament (PDL) cell cultures. Human PDL fibroblasts obtained from immature third molars of 11 patients were cultured and divided into two groups. The experimental group was cryopreserved and cultured after thawing. The control group was cultured without cryopreservation. A comparison was made between cryopreserved and control cells. To evaluate possible differences in the characteristics of the fibroblasts, the cells in both groups were tested for viability (membrane integrity), growth capacity and alkaline phosphatase (ALP) expression. The Wilcoxon test for paired comparison between cryopreserved and non-cryopreserved cells was performed for each characteristic. The results showed that membrane integrity of cells was not influenced by cryopreservation. There was no statistically significant difference in growth capacity between cryopreserved and control cells. Non-cryopreserved cells were slightly stronger positive for ALP, but the difference was not statistically significant. From these experiments it can be concluded that the observed parameters are not influenced by cryopreservation.

Evaluation of the antibacterial activity and toxicity of 2 new hydrogels: a pilot study.

Wound bed preparation remains a very important issue in wound healing. To promote the production of granulation tissue, it is necessary to remove necrotic tissue and to control infection. Necrotic tissue may be removed using a hydrogel preparation. Flaminal and Flaminal Hydro (Flen Pharma, Belgium) are 2 new hydroactive colloid gel dressings with state antibacterial properties. These properties are attributed to an enzymatic complex in their formulation. In the study described in this report, the antibacterial effects of Flaminal and Flaminal Hydro were confirmed in an in vitro as well as an in vivo setting. It was also demonstrated that Flaminal and Flaminal Hydro are not toxic to keratinocytes in vitro using an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test.

Double-blind placebo-controlled study of autologous transplanted epidermal cell suspensions for repigmenting vitiligo.

OBJECTIVES: To investigate the efficacy of epidermal noncultured cellular grafting in patients with vitiligo and the role of postinflammatory, spontaneous, or UV-induced pigmentation in obtaining repigmentation. DESIGN: A prospective, randomized, double-blind, placebo-controlled study. SETTING: Ambulatory patients in an institutional practice. Patients were followed up for 3 to 12 months. PATIENTS: A total of 33 paired, symmetrically distributed leukodermic lesions, all resistant to therapy, were observed in 28 patients. Nineteen patients appeared to have a stable vitiligo (group 1), whereas there was doubt about the stability of the disease in 9 patients (group 2). INTERVENTION: After laser ablation, a hyaluronic acid-enriched cellular graft was applied to 1 lesion while the paired lesion received placebo. Three weeks later all lesions were exposed to UV irradiation twice per week for approximately 2 months. MAIN OUTCOME MEASURES: Primarily, the percentage of repigmentation was assessed after 3, 6, and 12 months using a digital image analysis system. The repigmentation pattern was also evaluated after 1 and 3 months. RESULTS: A strongly significant difference between cellular grafts and placebo was observed after 3, 6, and 12 months (P<.001, P = .002, and P = .002, respectively). In group 1, repigmentation of at least 70% of the treated area was achieved in 55%, 57%, and 77% of the actively treated lesions 3, 6, and 12 months after treatment, whereas in group 2 repigmentation of at least 70% of the treated area was not observed at any time point. The repigmentation pattern was diffuse in 94% of the responding patients. CONCLUSIONS: After a strict preoperative selection for disease stability, transplantation resulted in repigmentation of at least 70% of the treated area in most actively treated vitiligo lesions. Repigmentation was primarily caused by the transplanted melanocytes.