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Study of in vitro capillary-like structures in psoriatic skin substitutes.

Ayata RE, Bouhout S, Auger M, Pouliot R - Biores Open Access (2014)

Bottom Line: Psoriatic fibroblasts and keratinocytes were isolated from psoriatic plaque biopsies, while healthy fibroblasts and keratinocytes, as well as microvascular endothelial cells, were isolated from healthy skin biopsies of cosmetic breast surgery.Also the semiquantification of capillary-like tubes (CLTs) was carried out with a 3D eagle eye view of substitutes, and more CLTs were observed in psoriatic substitutes.These results suggest that it is possible to observe 3D capillary-like structures in the self-assembled psoriatic skin substitutes, which could become a good in vitro testing model for anti-angiogenic drug research, and facilitate the study of this complex pathology, which links angiogenesis to its development.

View Article: PubMed Central - PubMed

Affiliation: Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Université Laval , Québec, Canada . ; Division of Regenerative Medicine, CHU de Québec Research Centre , Québec, Canada . ; Faculté de Pharmacie, Université Laval , Québec, Canada .

ABSTRACT
Angiogenesis is one of the important hallmarks of psoriasis. The extension of the superficial microvascular structure and activated pro-angiogenic mediators in psoriasis seem to be important factors involved in the pathology. According to the changes of superficial microvasculature in psoriatic lesions, anti-angiogenic treatment could be a promising therapeutic strategy for psoriasis. The aim of this study was to construct an in vitro vascularized psoriatic skin substitute for fundamental research. Psoriatic fibroblasts and keratinocytes were isolated from psoriatic plaque biopsies, while healthy fibroblasts and keratinocytes, as well as microvascular endothelial cells, were isolated from healthy skin biopsies of cosmetic breast surgery. Psoriatic and healthy skin substitutes with and without endothelial cells were produced using the self-assembly approach. Afterward the substitutes were examined by histology, immunofluorescence studies, and three-dimensional (3D) confocal microscopy. Histological analysis and immunofluorescence staining of specific markers for endothelial cells (von Willebrand, PECAM-1 [CD31], and VE-cadherin [CD144]) and basement membrane component (collagen IV) demonstrated that endothelial cells have the ability to form capillary-like tubes. Moreover, the 3D branched structure of the capillary-like structures and an eagle eye view of them were observed by confocal microscopy. Also the semiquantification of capillary-like tubes (CLTs) was carried out with a 3D eagle eye view of substitutes, and more CLTs were observed in psoriatic substitutes. These results suggest that it is possible to observe 3D capillary-like structures in the self-assembled psoriatic skin substitutes, which could become a good in vitro testing model for anti-angiogenic drug research, and facilitate the study of this complex pathology, which links angiogenesis to its development.

No MeSH data available.


Related in: MedlinePlus

Immunohistological localization of collagen IV (red) surrounded capillary-like tubes in the endothelialized psoriatic and healthy substitutes; ECs are labeled with CD31 (green). The last horizontal row indicates the collagen IV secretion of monolayer ECs under the same conditions as the substitutes (arrows indicate CLSs; scale bar=50 μm).
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f5: Immunohistological localization of collagen IV (red) surrounded capillary-like tubes in the endothelialized psoriatic and healthy substitutes; ECs are labeled with CD31 (green). The last horizontal row indicates the collagen IV secretion of monolayer ECs under the same conditions as the substitutes (arrows indicate CLSs; scale bar=50 μm).

Mentions: This last immunostaining study was done to determine the organizational and functional stability of CLSs in the substitutes. This was accomplished using anti-Coll IV, an important component of the BM of capillaries and the extracellular matrix that provides mechanical support against blood pressure. ECs secrete a subendothelial BM around the EC layer, which has an important role in the dilatation of capillaries and immune cell extravasation (indicated by arrows in Fig. 5). Additionally, it was proven that ECs were able to secrete their own matrix in vitro by monolayer studies (Fig. 5, horizontal bottom row).


Study of in vitro capillary-like structures in psoriatic skin substitutes.

Ayata RE, Bouhout S, Auger M, Pouliot R - Biores Open Access (2014)

Immunohistological localization of collagen IV (red) surrounded capillary-like tubes in the endothelialized psoriatic and healthy substitutes; ECs are labeled with CD31 (green). The last horizontal row indicates the collagen IV secretion of monolayer ECs under the same conditions as the substitutes (arrows indicate CLSs; scale bar=50 μm).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4215329&req=5

f5: Immunohistological localization of collagen IV (red) surrounded capillary-like tubes in the endothelialized psoriatic and healthy substitutes; ECs are labeled with CD31 (green). The last horizontal row indicates the collagen IV secretion of monolayer ECs under the same conditions as the substitutes (arrows indicate CLSs; scale bar=50 μm).
Mentions: This last immunostaining study was done to determine the organizational and functional stability of CLSs in the substitutes. This was accomplished using anti-Coll IV, an important component of the BM of capillaries and the extracellular matrix that provides mechanical support against blood pressure. ECs secrete a subendothelial BM around the EC layer, which has an important role in the dilatation of capillaries and immune cell extravasation (indicated by arrows in Fig. 5). Additionally, it was proven that ECs were able to secrete their own matrix in vitro by monolayer studies (Fig. 5, horizontal bottom row).

Bottom Line: Psoriatic fibroblasts and keratinocytes were isolated from psoriatic plaque biopsies, while healthy fibroblasts and keratinocytes, as well as microvascular endothelial cells, were isolated from healthy skin biopsies of cosmetic breast surgery.Also the semiquantification of capillary-like tubes (CLTs) was carried out with a 3D eagle eye view of substitutes, and more CLTs were observed in psoriatic substitutes.These results suggest that it is possible to observe 3D capillary-like structures in the self-assembled psoriatic skin substitutes, which could become a good in vitro testing model for anti-angiogenic drug research, and facilitate the study of this complex pathology, which links angiogenesis to its development.

View Article: PubMed Central - PubMed

Affiliation: Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Université Laval , Québec, Canada . ; Division of Regenerative Medicine, CHU de Québec Research Centre , Québec, Canada . ; Faculté de Pharmacie, Université Laval , Québec, Canada .

ABSTRACT
Angiogenesis is one of the important hallmarks of psoriasis. The extension of the superficial microvascular structure and activated pro-angiogenic mediators in psoriasis seem to be important factors involved in the pathology. According to the changes of superficial microvasculature in psoriatic lesions, anti-angiogenic treatment could be a promising therapeutic strategy for psoriasis. The aim of this study was to construct an in vitro vascularized psoriatic skin substitute for fundamental research. Psoriatic fibroblasts and keratinocytes were isolated from psoriatic plaque biopsies, while healthy fibroblasts and keratinocytes, as well as microvascular endothelial cells, were isolated from healthy skin biopsies of cosmetic breast surgery. Psoriatic and healthy skin substitutes with and without endothelial cells were produced using the self-assembly approach. Afterward the substitutes were examined by histology, immunofluorescence studies, and three-dimensional (3D) confocal microscopy. Histological analysis and immunofluorescence staining of specific markers for endothelial cells (von Willebrand, PECAM-1 [CD31], and VE-cadherin [CD144]) and basement membrane component (collagen IV) demonstrated that endothelial cells have the ability to form capillary-like tubes. Moreover, the 3D branched structure of the capillary-like structures and an eagle eye view of them were observed by confocal microscopy. Also the semiquantification of capillary-like tubes (CLTs) was carried out with a 3D eagle eye view of substitutes, and more CLTs were observed in psoriatic substitutes. These results suggest that it is possible to observe 3D capillary-like structures in the self-assembled psoriatic skin substitutes, which could become a good in vitro testing model for anti-angiogenic drug research, and facilitate the study of this complex pathology, which links angiogenesis to its development.

No MeSH data available.


Related in: MedlinePlus