Cultured human keratinocytes as a single cell suspension in fibrin glue combined with preserved dermal grafts enhance skin reconstitution in athymic mice full-thickness wounds

Horch, Raymund E.; Bannasch, Holger; Stark, G. Björn

doi:10.1007/s002380050196

Cited by 13 publications

(5 citation statements)

References 30 publications

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“…After mice were anesthetized with an intramuscular administration of ketamin hydrochloride (50 mg/kg, Yuhan Co., Seoul, Korea) and xylazine hydrochloride (5 mg/kg, Bayer Korea Ltd., Seoul, Korea), the full‐thickness skin wounds (1.5 × 1.5 cm 2 rectangular shape) were created on the back of athymic mice (BALB/c‐nu, 7 weeks old, female, SLC, Tokyo, Japan) 17. To minimize the migration of host skin cells from the wound margins and spontaneous wound contraction, the skins at the wound margins were burned using a cautery and fixed to adjacent muscle layers with nonresorbable 5‐0 nylon sutures (AILEE Co., Pusan, Korea) 17, 18. Dermal fibroblasts (approximately 10 8 cells/wound) and keratinocytes (approximately 7.5 × 10 6 cells/wound) cultured on PLGA microspheres were transplanted to the wounds (cell‐transplanted group, n =3) using a 1‐mL syringe without a needle.…”

Section: Methodsmentioning

confidence: 99%

“…17 To minimize the migration of host skin cells from the wound margins and spontaneous wound contraction, the skins at the wound margins were burned using a cautery and fixed to adjacent muscle layers with nonresorbable 5-0 nylon sutures (AILEE Co., Pusan, Korea). 17,18 Dermal fibroblasts (approximately 10 8 cells/wound) and keratinocytes (approximately 7.5 ϫ 10 6 cells/wound) cultured on PLGA microspheres were transplanted to the wounds (cell-transplanted group, n ϭ3) using a 1-mL syringe without a needle. After transplantation, the wounds were dressed with dressing materials, Tegaderm (3M Health Care, St. Paul, MN) and sterile cotton gauze, and firmly fixed using Coban, a selfadhesive wrap (3M Health Care).…”

Section: Transplantationmentioning

confidence: 99%

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Skin regeneration using keratinocytes and dermal fibroblasts cultured on biodegradable microspherical polymer scaffolds

et al. 2005

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Bioartificial skin sheet grafts have been utilized to treat large burns and chronic ulcers. However, the trypsinization step to harvest cultured skin grafts from culture dishes damages the cells by breaking the anchoring proteins and lowers their uptake ratio after transplantation. In addition, epidermal sheet grafts require a long fabrication period. To overcome these limitations, we utilized biodegradable poly(lactide-co-glycolide) (PLGA) microspheres as both cell culture matrix and transplantation vehicle of skin cells for skin regeneration in this study. This method could avoid the trypsinization step and have a relatively short preparation period. Human keratinocytes and dermal fibroblasts cultured on PLGA microspheres in spinner flasks proliferated by 3.0-fold and 9.4-fold, respectively, after 10 days. When both types of cells cultured on PLGA microspheres were reinoculated onto culture dishes, the cells migrated from the PLGA microspheres to the culture dish surface, grew, and formed a confluent cell layer within 5 days, showing the growth and migration abilities of the cells cultured on PLGA microspheres. Full-thickness skin wounds created on the back of athymic mice were either treated with transplantation of keratinocytes and dermal fibroblasts cultured on microspheres (cell-transplanted group), treated with PLGA microspheres alone (microsphere-implanted group), or covered with dressing materials without treatment (untreated group). Three weeks after the treatments, differentiated epithelium that stained positively for cytokeratin, a marker of epidermis, was observed in the cell-transplanted group, while the microsphere-implanted group and untreated group showed incomplete reepithelialization. Dermal regeneration with positive staining for vimentin, a marker of dermal fibroblast, was observed in the cell-transplanted group. Regenerated dermis with positive staining for vimentin was partly observed in the microsphere-implanted group and untreated group. These results suggest that transplantation of keratinocytes and dermal fibroblasts cultured on PLGA microspheres could be potentially useful as an alternative to bioartificial skin grafts for the treatment of skin wounds.

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Section: Methodsmentioning

confidence: 99%

Section: Transplantationmentioning

confidence: 99%

Skin regeneration using keratinocytes and dermal fibroblasts cultured on biodegradable microspherical polymer scaffolds

et al. 2005

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“…Recently, research efforts have shifted from multilayered differentiated cultured skin equivalents toward transplantation of: (1) single‐cell suspension of autologous cultured keratinocytes 28–32 or combined cultured keratinocytes and fibroblasts, 33 or (2) monolayers of autologous keratinocytes cultured on biodegradable 34,35 or synthetic membranes 36 …”

mentioning

confidence: 99%

Autologous cultured keratinocytes on porcine gelatin microbeads effectively heal chronic venous leg ulcers

Liu

Hafner

Dragieva

et al. 2004

Wound Repair Regeneration

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We have established a specific bioreactor microcarrier cell culture system using porcine gelatin microbeads as carriers to produce autologous keratinocytes on a large scale. Moreover, we have shown that autologous keratinocytes can be cultured on porcine collagen pads, thereby forming a single cell layer. The objective of this study was to compare efficacy and safety of autologous cultured keratinocytes on microbeads and collagen pads in the treatment of chronic wounds. Fifteen patients with recalcitrant venous leg ulcers were assigned to three groups in a single-center, prospective, uncontrolled study: five underwent a single treatment with keratinocyte monolayers on collagen pads (group 1); another five received a single grafting with keratinocyte-microbeads (group 2); and the last five received multiple, consecutive applications of keratinocyte-microbeads 3 days apart (group 3). All patients were followed for up to 12 weeks. By 12 weeks, there was a mean reduction in the initial wound area of 50, 83, and 97 percent in the three groups, respectively. The changes in wound size were statistically significant between the first and third groups (p= 0.0003). Keratinocyte-microbeads proved to be more effective than keratinocyte monolayers on collagen pads when the former were applied every 3 days. Rapid availability within 10-13 days after skin biopsy and easy handling represent particular advantages.

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“…Xenografts (tissue from another species) are more accessible than allografts. However, xenografts cannot be permanently re-vascularized from the wound so the tissue breaks down and sloughs off the wound [60].…”

Section: Conventional "Sheet Graft" (Cea) Skin Substitutesmentioning

confidence: 99%

Tissue engineering of cultured skin substitutes

et al. 2005

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Skin replacement has been a challenging task for surgeons ever since the introduction of skin grafts by Reverdin in 1871. Recently, skin grafting has evolved from the initial autograft and allograft preparations to biosynthetic and tissue-engineered living skin replacements. This has been fostered by the dramatically improved survival rates of major burns where the availability of autologous normal skin for grafting has become one of the limiting factors. The ideal properties of a temporary and a permanent skin substitute have been well defined. Tissue-engineered skin replacements: cultured autologous keratinocyte grafts, cultured allogeneic keratinocyte grafts, autologous/allogeneic composites, acellular biological matrices, and cellular matrices including such biological substances as fibrin sealant and various types of collagen, hyaluronic acid etc. have opened new horizons to deal with such massive skin loss. In extensive burns it has been shown that skin substitution with cultured grafts can be a life-saving measure where few alternatives exist. Future research will aim to create skin substitutes with cultured epidermis that under appropriate circumstances may provide a wound cover that could be just as durable and esthetically acceptable as conventional split-thickness skin grafts. Genetic manipulation may in addition enhance the performance of such cultured skin substitutes. If cell science, molecular biology, genetic engineering, material science and clinical expertise join their efforts to develop optimized cell culture techniques and synthetic or biological matrices then further technical advances might well lead to the production of almost skin like new tissue-engineered human skin products resembling natural human skin.

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Cultured human keratinocytes as a single cell suspension in fibrin glue combined with preserved dermal grafts enhance skin reconstitution in athymic mice full-thickness wounds

Cited by 13 publications

References 30 publications

Skin regeneration using keratinocytes and dermal fibroblasts cultured on biodegradable microspherical polymer scaffolds

Skin regeneration using keratinocytes and dermal fibroblasts cultured on biodegradable microspherical polymer scaffolds

Autologous cultured keratinocytes on porcine gelatin microbeads effectively heal chronic venous leg ulcers

Tissue engineering of cultured skin substitutes

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