Method for autologous single skin cell isolation for regenerative cell spray transplantation with non-cultured cells.

BACKGROUND: There is a therapeutic gap for patients with deep partial thickness wounds (Grade IIb) of moderate size that were initially not treated with split- or mesh grafting to avoid overgrafting, but developed delayed wound healing around two weeks after injury--at which time grafting is typically not indicated anymore. Delayed wound healing is often associated with esthetically unsatisfactory results and sometimes functional problems. An innovative cell isolation method for cell spray transplantation at the point of care, which eliminates cell culture prior to treatment, was implemented for this population of burn patients in our center. METHODS: Autologous skin cell spray transplantation was initiated by taking healthy skin. The dermal/epidermal layers were separated using enzymatic digestion with 40 min dispase application, followed by 15 min trypsin application for basal kerationcyte isolation, 7 min cell washing by centrifugation, followed by transferring the cells for spraying into Ringer lactate solution. The procedure was performed on site in a single session immediately following the biopsy. After sharp wound debridement, cells were immediately transplanted by deposition with a cell sprayer for even distribution of the cell suspension. RESULTS AND CONCLUSIONS: Eight patients were treated (mean age 30.3 years, mean burn total body surface area 14%, mean Abbreviated Burn Severity Index (5 points). The mean time to complete re-epithelialization was 12.6 days. All patients exhibited wound healing with improved esthetic and functional quality. Our initial experience for the use of non-cultured cells using a two-enzyme approach with cell washing suggests shortened time for wound closure, suggesting that the method may potentially avoid longer-term complications.

Sprayed cultured epithelial autografts for deep dermal burns of the face and neck.

The objective of this study was the assessment of clinical results after sprayed application of cultured epithelial autograft (CEA) suspensions onto deep dermal burn wounds of the face and neck. Nineteen patients with deep dermal burns of the face and neck were included into a prospective study. The average total body surface area burn was 15.1% (7%-46%; median: 13%). The average Abbreviated Burn Severity Index (ABSI) was 6.7 points (4-12 points; median: 7 points). The application of sprayed CEA suspension was performed onto an average body surface area of 2% (0.5-5%; median: 2%). Thirteen patients were recruited for clinical follow-up after an average of 10 months (3-18 months). The average Vancouver Scar Scale score at follow-up was 2.4 +/- 2.2 points (range, 0-8 points), and the average Donnersmarck and Hörbrand score was 9.3 +/- 6.8 points (range, 0-22). Four patients had less than 9 months' follow-up. Excluding these patients from the analysis resulted in an average Vancouver Scar Scale score of 1.3 +/- 0.9 points (range, 0-3 points) and an average Donnersmarck and Hörbrand score of 8.0 +/- 7.4 points (range 0-22) for the remaining 9 patients.Our data show that enzymatic and careful surgical debridement and consecutive application of CEA suspensions using a spray technique results in excellent cosmetic outcomes compared with any other method.