Cultured autologous keratinocytes in fibrin glue suspension, exclusively and combined with STS-allograft (preliminary clinical and histological report of a new technique).

The use of cultured epidermal cell sheets has become a recognized method for the coverage of extensive burns. The disadvantages are a long time-lag until the cells are available, the fragility and difficult handling of the grafts, an unpredictable 'take' and extremely high costs. In three patients with deep partial and full skin thickness burns we have applied cultured autologous keratinocytes suspended in fibrin glue. In two of these patients the keratinocyte culture in the fibrin matrix (KFGS) was overgrafted with allogeneic, glycerine-preserved split thickness cadaver skin. The area thus covered ranged from 3 to 15 per cent TBSA. Cultured grafts were available between 2.5 and 3 weeks. The non-confluent cells developed a continuous epithelial layer within the 4 days until the first dressing change. Histological examination showed a stratified neoepidermis. Clinically the new skin had satisfactory stability and mechanical quality. The epidermis of the allogeneic overgrafts desquamated within a few days without signs of inflammation, but there are indications that the STS-allograft dermis is at least partly integrated into the new skin and may serve as a scaffold for the grafted cell culture. The fibrin glue matrix seems to give sufficient adherence stability to keratinocytes that are grafted in an actively proliferating state. Further advantages are the easy repetition and application, as well as a reduction in operating time and costs in these severely injured patients.

Localization of adducin in epidermis.

On the cytoplasmic side of the plasma membrane of erythrocytes there is a dense protein filament matrix that maintains the shape of the cells. The main constituents of this system, actin and spectrin, which have also been detected in keratinocytes and fibroblasts, are known to be linked in erythrocytes in a network structure by additional proteins such as band 4.1 and adducin. The interaction between actin and spectrin, mediated by adducin, is regulated by calmodulin and protein kinase C. Because we have previously found adducin in cultured keratinocytes, we investigated epidermis by immunochemical techniques. We found adducin to be localized at cell-cell contact sites in epidermis using affinity-purified antibodies against human erythrocyte adducin. Immunofluorescence of epidermis revealed an intense fluorescence in the basal layer, whereas stratum spinosum and stratum granulosum showed moderate staining. There was intense staining at sites of cell-cell contact in cultured human keratinocytes. Immunoblot analysis indicated the presence of adducin polypeptides of 103 kd and 97 kd in epidermis, but in cultured keratinocytes only the higher molecular weight form could be detected. This study indicates adducin, a regulatory protein in erythrocytes, is also present in epidermis. Its localization suggests that it may be involved in the formation of the microfilament matrix of the membrane skeleton at cell-cell contact sites.