Wound closure with human keratinocytes cultured on a polyurethane dressing overlaid on a cultured human dermal replacement

Rennekampff, Hans Oliver; Hansbrough, John F.; Kiessig, Verena; AbiEzzi, Salwan S.; Woods, Virgil L.

doi:10.1016/s0039-6060(96)80235-2

Cited by 53 publications

(24 citation statements)

References 15 publications

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“…Other researchers have shown that synthetic materials may also serve as a carrier to deliver confluent keratinocyte cell layers to wounds. 37 Unlike with conventiona l composite skin grafts in this system no time is required to revascularize a matrix in order to reestablish nutrient provision to the transplanted cells. This ensures a high viability of the transferred cells in wound environm ent.…”

Section: Discussionmentioning

confidence: 99%

“…To avoid any barrier function of the carrier, we transplanted composite grafts with the keratinocyte layer in an "upside-down" direction towards the wound bed, so that the collagen component serves as a carrier and as a biological dressing on top. In contrast to studies previously published by others, 37 we used subconfluent monolayers of cultured human keratinocytes instead of confluent layers or multilayered sheet grafts. The aim of this study was to prove the feasibility of this approach utilizing keratinocyte collagen membrane grafts (KCMG) "upsidedown" grafts onto full-thickne ss nude mice wounds to reconstitute the epithelium .…”

Section: Introductionmentioning

confidence: 99%

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Cultured Human Keratinocytes on Type I Collagen Membranes to Reconstitute the Epidermis

Horch

Debus²,

Wagner³

et al. 2000

Tissue Engineering

114

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The development of new techniques and modifications to overcome some of the disadvantages in cultured keratinocyte grafting has been motivated by several well-known drawbacks in the use of cultured epithelial autografts such as long culture periods, lack of adherence, difficulty in handling, lack of dermal substrates, and high costs. Two recent insights have influenced further research. On the one hand, it has been shown that the use of undifferentiated proliferative cells in fibrin glue suspensions is effective in epithelial reconstitution. On the other hand, the enzymatic release of cells from the culture surfaces is a critical step leading to at least temporary destruction of anchoring structures of the cultured cells. In this study, we tried to combine these two aspects in an attempt to modify common modalities of keratinocyte transplantation. To avoid dispase dissolving of the cultured cells, keratinocytes were seeded onto bovine collagen type I membranes without feeder layers and under serum-free culture conditions. Subconfluent monolayers of cultured human keratinocytes were transplanted as an upside-down graft on collagen membranes (keratinocyte collagen membrane grafts [KCMG], n = 12) after 3 days of culture or as membrane grafts alone (n = 12) onto standard nude mice full-thickness wounds. Fully differentiated epidermis was found at 21 days after grafting KCMG with persistence of human keratinocytes. This study demonstrates that upside-down grafts of undifferentiated monolayers of keratinocytes on non-cross-linked bovine type I collagen membranes do lead to an early reconstitution of multilayered squamous epithelium with enhanced wound healing compared to the control group. The upside down KCMG grafting technique is able to transfer actively proliferative keratinocytes and simplifies the application compared to conventional epithelial sheet grafting.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cultured Human Keratinocytes on Type I Collagen Membranes to Reconstitute the Epidermis

Horch

Debus²,

Wagner³

et al. 2000

Tissue Engineering

114

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“…[6,7] For example, the commercially available product Epicel 1 uses polyurethane membranes as scaffold for the cultivation and transfer of keratinocytes to the wound bed. [8] Adhesion, growth and function of cells on biomedical materials are dependent on the surface properties of biomaterials.…”

Section: Introductionmentioning

confidence: 99%

Poly(ether imide) Membranes Modified with Poly(ethylene imine) as Potential Carriers for Epidermal Substitutes

Trimpert

Boese

Albrecht

et al. 2006

Macromolecular Bioscience

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Poly(ether imide) (PEI) membranes were modified with a linear low-molecular weight (PETIM_0.6) and a branched high-molecular weight poly(ethylene imine) (PETIM_60). The membrane surfaces became more hydrophilic and the zeta potentials were shifted from negative to positive zeta values after immobilisation of both PETIM. These measurements also indicated the presence of a swollen surface layer in the case of PETIM_60, while a regular structuring of the surface was observed with scanning force microscopy for PETIM_0.6. A human keratinocyte cell line HaCaT was cultured on the different membranes. It was found that HaCaT cell growth was stimulated by PETIM_0.6. Cells reached earlier confluence on this substratum, while their growth was inhibited on a PEI membrane modified with PETIM_60, which makes PEI membranes modified with PETIM_0.6 a promising material for in vitro culture of epidermal transplants.

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“…, polyurethane membranes(32), porous synthetic carriers (44), and aerosolThe aim of our study was to isolate and to culture sufficient numbers of keratinocytes that are still able to sprays (24). Such keratinocyte delivery systems are easier to handle than the epidermal sheets.For the coverage regenerate an epidermis in a short time frame without the need of mouse feeder layer cells and FCS.…”

mentioning

confidence: 99%

Culture of Keratinocytes for Transplantation without the Need of Feeder Layer Cells

et al. 2007

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Patients with large burn wounds have a limited amount of healthy donor skin. An alternative for the autologous skin graft is transplantation with autologous keratinocytes. Conventionally, the keratinocytes are cultured with mouse feeder layer cells in medium containing fetal calf serum (FCS) to obtain sufficient numbers of cells. These xenobiotic materials can be a potential risk for the patient. The aim of the present study was to investigate if keratinocytes could be expanded in culture without the need of a feeder layer and FCS. Keratinocytes were cultured on tissue culture plastic with or without collagen type IV coating in medium containing Ultroser G (serum substitute) and keratinocyte growth factor (KGF). An in vitro skin equivalent model was used to examine the capacity of these cells to form an epidermis. Keratinocytes in different passages (P2, P4, and P6) and freshly isolated cells were studied. Keratinocytes grown on collagen type IV were able to form an epidermis at higher passage numbers than cells grown in the absence of collagen type IV (P4 and P2, respectively). In both cases the reconstructed epidermis showed an increased expression of Ki-67, SKALP, involucrin, and keratin 17 compared to normal skin. Only 50,000 keratinocytes grown on collagen type IV in P4 were needed to form 1 cm2 epidermis, whereas 150,000 of freshly isolated keratinocytes were necessary. Using this culture technique sufficient numbers of keratinocytes, isolated from 1 cm2 skin, were obtained to cover 400 cm2 of wound surface in 2 weeks. The results show that keratinocytes can be cultured without the need of a fibroblast feeder layer and FCS and that these cells are still able to create a fully differentiated epidermis. This culture technique can be a valuable tool for the treatment of burn wounds and further development of tissue engineered skin.

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Wound closure with human keratinocytes cultured on a polyurethane dressing overlaid on a cultured human dermal replacement

Cited by 53 publications

References 15 publications

Cultured Human Keratinocytes on Type I Collagen Membranes to Reconstitute the Epidermis

Cultured Human Keratinocytes on Type I Collagen Membranes to Reconstitute the Epidermis

Poly(ether imide) Membranes Modified with Poly(ethylene imine) as Potential Carriers for Epidermal Substitutes

Culture of Keratinocytes for Transplantation without the Need of Feeder Layer Cells

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