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Biodegradable Gelatin Microcarriers Facilitate Re-Epithelialization of Human Cutaneous Wounds - An In Vitro Study in Human Skin.

Lönnqvist S, Rakar J, Briheim K, Kratz G - PLoS ONE (2015)

Bottom Line: Tissue sections showed incorporation of wound edge keratinocytes into the microcarriers and thicker neoepidermis in wounds treated with microcarriers.Air-lifting of wounds enhanced stratification in control wounds as well as wounds with CultiSpher-S.We conclude that the CultiSpher-S microcarriers can function as tissue guiding scaffold for re-epithelialization of cutaneous wounds.

View Article: PubMed Central - PubMed

Affiliation: Division of Experimental Plastic Surgery, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University, Linköping, Sweden.

ABSTRACT
The possibility to use a suspended tridimensional matrix as scaffolding for re-epithelialization of in vitro cutaneous wounds was investigated with the aid of a human in vitro wound healing model based on viable full thickness skin. Macroporous gelatin microcarriers, CultiSpher-S, were applied to in vitro wounds and cultured for 21 days. Tissue sections showed incorporation of wound edge keratinocytes into the microcarriers and thicker neoepidermis in wounds treated with microcarriers. Thickness of the neoepidermis was measured digitally, using immunohistochemical staining of keratins as epithelial demarcation. Air-lifting of wounds enhanced stratification in control wounds as well as wounds with CultiSpher-S. Immunohistochemical staining revealed expression of keratin 5, keratin 10, and laminin 5 in the neoepidermal component. We conclude that the CultiSpher-S microcarriers can function as tissue guiding scaffold for re-epithelialization of cutaneous wounds.

No MeSH data available.


Immunohistochemical staining.(A) Immunohistological staining against keratin 10 (red) and keratin 5 (green) in neoepidermis with incorporated microcarriers and compared to (B) normal control skin. (C) Immunohistological staining against pancytokeratin (red) showing microcarrier populated by keratinocytes. (D) Immunohistological staining against laminin 5 (green) in neoepidermis with microcarriers, (E) unwounded skin, and (F) neoepidermis of control wound. Nuclear staining with 4, 6-diamidino-2-phenylindole (DAPI). All scale bars = 100 μm.
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pone.0128093.g003: Immunohistochemical staining.(A) Immunohistological staining against keratin 10 (red) and keratin 5 (green) in neoepidermis with incorporated microcarriers and compared to (B) normal control skin. (C) Immunohistological staining against pancytokeratin (red) showing microcarrier populated by keratinocytes. (D) Immunohistological staining against laminin 5 (green) in neoepidermis with microcarriers, (E) unwounded skin, and (F) neoepidermis of control wound. Nuclear staining with 4, 6-diamidino-2-phenylindole (DAPI). All scale bars = 100 μm.

Mentions: Immunohistological staining revealed expression of keratin 5 and keratin 10 in the neoepidermis of wounds with microcarriers (Fig 3A) in compartments corresponding to mature epidermis of normal skin (Fig 3B). Local expression of laminin 5 in keratinocytes on the surface of the microcarriers (Fig 3D) was detected, compared to the uniform expression in unwounded skin (Fig 3E) and neoepidermis of control wound (Fig 3F). No increase in Ki-67 expression at wound edges could be detected (not shown). Confocal imaging of pancytokeratin staining of wounds with microcarriers showed the CultiSpher-S completely populated by keratinocytes (Fig 3C).


Biodegradable Gelatin Microcarriers Facilitate Re-Epithelialization of Human Cutaneous Wounds - An In Vitro Study in Human Skin.

Lönnqvist S, Rakar J, Briheim K, Kratz G - PLoS ONE (2015)

Immunohistochemical staining.(A) Immunohistological staining against keratin 10 (red) and keratin 5 (green) in neoepidermis with incorporated microcarriers and compared to (B) normal control skin. (C) Immunohistological staining against pancytokeratin (red) showing microcarrier populated by keratinocytes. (D) Immunohistological staining against laminin 5 (green) in neoepidermis with microcarriers, (E) unwounded skin, and (F) neoepidermis of control wound. Nuclear staining with 4, 6-diamidino-2-phenylindole (DAPI). All scale bars = 100 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4464648&req=5

pone.0128093.g003: Immunohistochemical staining.(A) Immunohistological staining against keratin 10 (red) and keratin 5 (green) in neoepidermis with incorporated microcarriers and compared to (B) normal control skin. (C) Immunohistological staining against pancytokeratin (red) showing microcarrier populated by keratinocytes. (D) Immunohistological staining against laminin 5 (green) in neoepidermis with microcarriers, (E) unwounded skin, and (F) neoepidermis of control wound. Nuclear staining with 4, 6-diamidino-2-phenylindole (DAPI). All scale bars = 100 μm.
Mentions: Immunohistological staining revealed expression of keratin 5 and keratin 10 in the neoepidermis of wounds with microcarriers (Fig 3A) in compartments corresponding to mature epidermis of normal skin (Fig 3B). Local expression of laminin 5 in keratinocytes on the surface of the microcarriers (Fig 3D) was detected, compared to the uniform expression in unwounded skin (Fig 3E) and neoepidermis of control wound (Fig 3F). No increase in Ki-67 expression at wound edges could be detected (not shown). Confocal imaging of pancytokeratin staining of wounds with microcarriers showed the CultiSpher-S completely populated by keratinocytes (Fig 3C).

Bottom Line: Tissue sections showed incorporation of wound edge keratinocytes into the microcarriers and thicker neoepidermis in wounds treated with microcarriers.Air-lifting of wounds enhanced stratification in control wounds as well as wounds with CultiSpher-S.We conclude that the CultiSpher-S microcarriers can function as tissue guiding scaffold for re-epithelialization of cutaneous wounds.

View Article: PubMed Central - PubMed

Affiliation: Division of Experimental Plastic Surgery, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University, Linköping, Sweden.

ABSTRACT
The possibility to use a suspended tridimensional matrix as scaffolding for re-epithelialization of in vitro cutaneous wounds was investigated with the aid of a human in vitro wound healing model based on viable full thickness skin. Macroporous gelatin microcarriers, CultiSpher-S, were applied to in vitro wounds and cultured for 21 days. Tissue sections showed incorporation of wound edge keratinocytes into the microcarriers and thicker neoepidermis in wounds treated with microcarriers. Thickness of the neoepidermis was measured digitally, using immunohistochemical staining of keratins as epithelial demarcation. Air-lifting of wounds enhanced stratification in control wounds as well as wounds with CultiSpher-S. Immunohistochemical staining revealed expression of keratin 5, keratin 10, and laminin 5 in the neoepidermal component. We conclude that the CultiSpher-S microcarriers can function as tissue guiding scaffold for re-epithelialization of cutaneous wounds.

No MeSH data available.