Direct Comparison of a Cultured Composite Skin Substitute Containing Human Keratinocytes and Fibroblasts to an Epidermal Sheet Graft Containing Human Keratinocytes on Athymic Mice

Cooper, Matthew; Andree, Christoph; Hansbrough, John F.; Zapata-Sirvent, Ramon L.; Spielvogel, Richard L.

doi:10.1111/1523-1747.ep12371700

Cited by 93 publications

(82 citation statements)

References 42 publications

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“…6,26,30,[32][33][34] In our study, the results demonstrate that the presence of a healthy and intact epithelial layer on the EVPOME enhanced vascular infiltration into the underlying dermis when compared with the control samples, AlloDerm without epithelium, which showed less fibroblastic infiltration and vessel lumen formation (Table 2). In addition, the different patterns of vascular ingrowth seen within the dermal layer of the three types of EVPOME, D4E, D11E, and D18E, are thought to be related to the number and stage of differentiation of the overlying keratinocytes.…”

Section: Discussionmentioning

confidence: 71%

“…[22][23][24][25] In addition, it appears important that a human dermal matrix retain a basement membrane structure as a scaffold to as-TRANSPLANTATION ORAL MUCOSA EQUIVALENTS sist in the optimal growth and differentiation of keratinocytes. 23,26 AlloDerm with epithelium, EVPOME, showed marked remodeling of the collagenous bundles in comparison with AlloDerm without epithelium. This eventually progressed to a point of contracture of the EVPOME at 21 days postgrafting.…”

Section: Discussionmentioning

confidence: 93%

“…5,6 To overcome these adverse events, artificial dermal substitutes have been used to assist in keratinocyte growth. [7][8][9][10][11][12] One dermal analog used for treatment of full-thickness wounds is an acellular human cadaver dermis, AlloDerm (LifeCell, Branchburg, NJ).…”

Section: Introduction Smentioning

confidence: 99%

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Evaluation of Transplanted Tissue-Engineered Oral Mucosa Equivalents in Severe Combined Immunodeficient Mice

et al. 2003

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The aim of this study was to determine the optimal stage of development at which transplant human ex vivo-produced oral mucosa equivalents (EVPOMEs) in vivo. EVPOMEs were generated in a serum-free culture system, without the use of an irradiated xenogeneic feeder layer, by seeding human oral keratinocytes onto a human cadaveric dermal equivalent, AlloDerm. EVPOMEs were cultured for 4 days submerged and then for 7 or 14 days at an air-liquid interface to initiate stratification before transplantation into SCID mice. AlloDerm, without epithelium, was used as a control. Mice were killed on days 3, 10, and 21 posttransplantation. Epithelium of the transplanted EVPOMEs was evaluated with the differentiation marker keratin 10/13. Dermal microvessel ingrowth was determined by immunohistochemistry with a mouse vascular marker, lectin binding from Triticum vulgaris. The presence and stratification of the epithelium were correlated with revascularization of the underlying dermis. The microvessel density of AlloDerm without epithelium was less than that of EVPOMEs with an epithelial layer. Microvessel density of the dermis varied directly with the degree of epithelial stratification of the EVPOMEs. The EVPOMEs cultured at an air-liquid interface for 7 days had the optimal balance of neoangiogenesis and epithelial differentiation necessary for in vivo grafting. 163

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Section: Discussionmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 93%

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Evaluation of Transplanted Tissue-Engineered Oral Mucosa Equivalents in Severe Combined Immunodeficient Mice

et al. 2003

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“…The athymic mouse has been the preferred model to study the wound healing capability of human skin substitutes (21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33), because of its immunologic tolerance of xenograft and the limited scarring that occurs during healing. ESS grafts fabricated with keratinocytes harvested from the Kerator had well keratinized and desquamating surfaces after two weeks following surgery (Figure 3).…”

Section: Discussionmentioning

confidence: 99%

Wound Healing on Athymic Mice With Engineered Skin Substitutes Fabricated with Keratinocytes Harvested from an Automated Bioreactor

Kalyanaraman

Boyce

2009

Journal of Surgical Research

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The Kerator is a computer controlled bioreactor for the automated culture and harvest of keratinocytes that can reduce labor and materials involved in the fabrication of engineered skin substitutes (ESS). Previous studies have shown that Kerator is comparable to tissue culture flasks by keratinocyte confluence during culture, clonogenic potential of harvested keratinocytes and microanatomy, cell viability and surface hydration of ESS fabricated with the harvested keratinocytes. In this study, the Kerator and tissue culture flasks were further compared by keratinocyte proliferation in vitro and wound healing after transplantation of ESS to athymic mice. The number of BrdU positive keratinocytes in ESS fabricated with keratinocytes harvested from Kerator after 2 weeks of in vitro maturation was 34 ± 3 per high power field (hpf) (mean ± SEM), which was not significantly different from that fabricated with keratinocytes harvested from flasks (34 ± 1.5 per hpf). Percentage original wound area 6 weeks after surgery of ESS fabricated with keratinocytes from the Kerator was 36 ± 3.3%, which was not significantly different from that of ESS fabricated with keratinocytes from flasks (30 ± 4.3%). In both cases, 78% (7 of 9) mice transplanted were positive for engraftment of human keratinocytes by direct immunofluoresence for HLA-ABC antigens. These results further confirm that the ESS fabricated with keratinocytes harvested from Kerator and flasks are equivalent in vitro and in vivo. Therefore, use of Kerator for large scale production of ESS can lead to increased availability at reduced cost while maintaining ESS quality for grafting.

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“…In animal models, C-GAG grafts containing keratinocytes, melanocytes, and fibroblasts can regenerate an epidermis which inhibits wound contraction and re-pigments over time (Boyce et al, , 1993aHarriger et al, 1995). In a comparison study on full-thickness wounds of athymic mice, the collagen-GAG composite skin graft appeared to be superior to epidermal sheets in the rate of "take" and histologic appearance, although, in contrast to epidermal sheets in a short-term study (a maximum of 42 days), composite skin grafts formed a dermo-epidermal junction very well (Cooper et al, 1993). Detailed sequential studies for three months in three patients showed good matrix incorporation into the wounds, with a well-formed basement membrane, normal-appearing anchoring fibrils, and expression of skin antigens (Boyce et al, 1993b).…”

Section: (All) In Vitro Tissue Engineering Of the Dermismentioning

confidence: 99%

Tissue Engineering of Skin

Pomahac¹,

Svensjö²,

Yao³

et al. 1998

Critical Reviews in Oral Biology & Medicine

103

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The skin plays a crucial role in protecting the integrity of the body's internal milieu. The loss of this largest organ is incompatible with sustained life. In reconstructive surgery or burn management, substitution of the skin is often necessary. In addition to traditional approaches such as split-or full-thickness skin grafts, tissue flaps and free-tissue transfers, skin bioengineering in vitro or in vivo has been developing over the past decades. It applies the principles and methods of both engineering and life sciences toward the development of substitutes to restore and maintain skin structure and function. Currently, these methods are valuable alternatives or complements to other techniques in reconstructive surgery. This review article deals with the evolution and current approaches to the development of in vitro and in vivo epidermis and dermis.

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Direct Comparison of a Cultured Composite Skin Substitute Containing Human Keratinocytes and Fibroblasts to an Epidermal Sheet Graft Containing Human Keratinocytes on Athymic Mice

Cited by 93 publications

References 42 publications

Evaluation of Transplanted Tissue-Engineered Oral Mucosa Equivalents in Severe Combined Immunodeficient Mice

Evaluation of Transplanted Tissue-Engineered Oral Mucosa Equivalents in Severe Combined Immunodeficient Mice

Wound Healing on Athymic Mice With Engineered Skin Substitutes Fabricated with Keratinocytes Harvested from an Automated Bioreactor

Tissue Engineering of Skin

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