Active wound dressing with artificial capillaries for temporary wound irrigation and skin cell supply.

Medical treatment of burns and chronic wounds remains a challenge. We discussed a therapy concept that combines skin cell spray transplantation with a novel wound dressing based on artificial hollow fiber membrane capillaries. In skin cell-based therapy development, autologous skin progenitor cells are isolated from a healthy skin area and sprayed onto the wound. A medical device was introduced that uses perfused capillaries, known from clinical plasma separation, as a temporarily applied extracorporeal wound capillary bed. The functions of the dressing are comparable with those of dialysis; the capillaries, however, are applied externally onto the wound. Perfusion with a clinical peripheral nutrition and buffer solution can provide wound irrigation, wound debris removal, cell nutrition, pH regulation, and electrolyte balance while potentially serving to address delivery of regenerative factors and antibiosis. An innovative active skin wound dressing that provides cell support and stimulates regeneration by wound irrigation is discussed.

Feasibility study of an active wound dressing based on hollow fiber membranes in a porcine wound model.

INVESTIGATION: A novel active wound dressing (AWD) concept based on a microporous hollow fiber membrane network was investigated in an animal model. It provides a local solution-perfused environment for regenerative cell nutrition, wound irrigation, debris removal, electrolyte balancing, pH regulation, and topical antibiosis. The device is capable of supplying soluble factors, as tested experimentally for the recombinant human growth and differentiation factor-5 (rhGDF-5). METHODS: Following in vitro studies for rhGDF-5 using primary human keratinocytes and dermal fibroblasts, we employed a porcine partial thickness wound model with five distinct wounds on each back of n=8 pigs. Four wound groups were perfused differently over 9 days and compared with a negative control wound without perfusion: (1) 1% trehalose solution, pH 5.5; (2) rhGDF-5 (150 ng/ml) in 1% trehalose solution, pH 5.5; (3) nutrition solution; and (4) rhGDF-5 (150 ng/ml) in nutrition solution with 1% trehalose, pH 5.5. RESULTS: Promoted wound healing was observed within group 1 and more pronounced within group 2. Groups 3 and 4, with nutrition solution, showed significant adverse effects on wound healing (p<0.05). CONCLUSIONS: The investigated AWD concept appears to be an interesting therapeutic tool to study further wound healing support. Additionally, topical application of rhGDF-5 could be promising.

Phenotypical characterization of 6-21-week gestational age human dermis and epidermal cell isolation methods for in vitro studies on epidermal progenitors.

UNLABELLED: Cell banked epidermal skin progenitor cells have the potential to provide an "off-the-freezer" product. Such cells may provide a skin donor area-independent cell-spray grafting therapy for the treatment of burns. We first characterized fetal skin samples of gestational ages ranging from 6 to 21 weeks. As the results suggest that the phenotypic differentiation occurs after 10 weeks, which may complicate follow-up in vitro studies, we developed and compared different cell isolation techniques for human fetal skin-derived epithelial cells from tissue ages 6 to 9 weeks. We initially screened seven methods of characterization, concluding that two methods warranted further investigation: incubating the epidermal tissue in Petri-dishes with culture medium for spontaneous cell outgrowth, and wiping the epidermal tissue onto a dry Petri-dish culture surface followed by adding culture medium. Non-controllable culture contamination with dermal cells was the reason for excluding the other five methods. The results suggest that epidermal cells can be isolated from tissue exhibiting a single homogeneous layer of CK15(+) basal keratinocytes up to week 9. At later gestational ages, the ongoing skin differentiation results in a multi-layer basal structure and progenitors associated with the hair bulb would have to be considered. Spraying the resulting cells with a clinical spray device was successfully demonstrated in an in vitro model. CONCLUSION: Gestational age 6-9 weeks epidermal human fetal skin cells from the basal layer can be reproducibly isolated and transferred into culture for studies on the development of skin cell transplantation therapies.

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.