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LPS-Stimulated Human Skin-Derived Stem Cells Enhance Neo-Vascularization during Dermal Regeneration.

Kisch T, Weber C, Rapoport DH, Kruse C, Schumann S, Stang FH, Siemers F, Matthießen AE - PLoS ONE (2015)

Bottom Line: High numbers of adult stem cells are still required to improve the formation of new vessels in scaffolds to accelerate dermal regeneration.Results showed that LPS-activated SDSC significantly enhanced vascularization of the scaffolds, compared to unstimulated stem cells in vivo.Our results suggest that combining activated stem cells and a dermal substitute is a promising option to enhance vascularization in scaffold-mediated dermal regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Plastic Surgery and Hand Surgery, University of Lübeck, Lübeck, Germany.

ABSTRACT
High numbers of adult stem cells are still required to improve the formation of new vessels in scaffolds to accelerate dermal regeneration. Recent data indicate a benefit for vascularization capacity by stimulating stem cells with lipopolysaccharide (LPS). In this study, stem cells derived from human skin (SDSC) were activated with LPS and seeded in a commercially available dermal substitute to examine vascularization in vivo. Besides, in vitro assays were performed to evaluate angiogenic factor release and tube formation ability. Results showed that LPS-activated SDSC significantly enhanced vascularization of the scaffolds, compared to unstimulated stem cells in vivo. Further, in vitro assays confirmed higher secretion rates of proangiogenic as well as proinflammatoric factors in the presence of LPS-activated SDSC. Our results suggest that combining activated stem cells and a dermal substitute is a promising option to enhance vascularization in scaffold-mediated dermal regeneration.

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Related in: MedlinePlus

SDSC-seeded scaffolds.Cell matrix interactions in the scaffold (Autofluorescence/green) were evaluated 4 hours after cell seeding. Cells are presented in blue (DNA: 40,6-diamidino-2-phenylindole (DAPI)) and red (F-actin: phalloidin). After 7 days a homogeneous distribution of SDSC in the scaffold could be shown (A). Cell metabolic activity in the scaffold was evaluated for different time points by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The metabolic activity remained significantly unchanged after 7 days of culture (white bars: LPS-stimulated SDSC; grey bars: unstimulated SDSC). 4 days later a significant difference in metabolic activity was found (B).
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pone.0142907.g002: SDSC-seeded scaffolds.Cell matrix interactions in the scaffold (Autofluorescence/green) were evaluated 4 hours after cell seeding. Cells are presented in blue (DNA: 40,6-diamidino-2-phenylindole (DAPI)) and red (F-actin: phalloidin). After 7 days a homogeneous distribution of SDSC in the scaffold could be shown (A). Cell metabolic activity in the scaffold was evaluated for different time points by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The metabolic activity remained significantly unchanged after 7 days of culture (white bars: LPS-stimulated SDSC; grey bars: unstimulated SDSC). 4 days later a significant difference in metabolic activity was found (B).

Mentions: Since SDSC are meant to be used in an in vivo wound healing model, their growth and stimulation ability in a collagen matrix was analyzed. Therefore, SDSC were cultured in 6 mm pieces of Integra matrix placed on 12-well plates. Scaffolds were incubated at 37°C for improving the attachment of cells to the matrix. Already 4 h post seeding, Phalloidin and DAPI staining visualized the interaction of SDSC with the surrounding matrix. When cultured for 7 days, a homogeneous distribution of viable cells became visible within the matrix (Fig 2A).


LPS-Stimulated Human Skin-Derived Stem Cells Enhance Neo-Vascularization during Dermal Regeneration.

Kisch T, Weber C, Rapoport DH, Kruse C, Schumann S, Stang FH, Siemers F, Matthießen AE - PLoS ONE (2015)

SDSC-seeded scaffolds.Cell matrix interactions in the scaffold (Autofluorescence/green) were evaluated 4 hours after cell seeding. Cells are presented in blue (DNA: 40,6-diamidino-2-phenylindole (DAPI)) and red (F-actin: phalloidin). After 7 days a homogeneous distribution of SDSC in the scaffold could be shown (A). Cell metabolic activity in the scaffold was evaluated for different time points by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The metabolic activity remained significantly unchanged after 7 days of culture (white bars: LPS-stimulated SDSC; grey bars: unstimulated SDSC). 4 days later a significant difference in metabolic activity was found (B).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4643997&req=5

pone.0142907.g002: SDSC-seeded scaffolds.Cell matrix interactions in the scaffold (Autofluorescence/green) were evaluated 4 hours after cell seeding. Cells are presented in blue (DNA: 40,6-diamidino-2-phenylindole (DAPI)) and red (F-actin: phalloidin). After 7 days a homogeneous distribution of SDSC in the scaffold could be shown (A). Cell metabolic activity in the scaffold was evaluated for different time points by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The metabolic activity remained significantly unchanged after 7 days of culture (white bars: LPS-stimulated SDSC; grey bars: unstimulated SDSC). 4 days later a significant difference in metabolic activity was found (B).
Mentions: Since SDSC are meant to be used in an in vivo wound healing model, their growth and stimulation ability in a collagen matrix was analyzed. Therefore, SDSC were cultured in 6 mm pieces of Integra matrix placed on 12-well plates. Scaffolds were incubated at 37°C for improving the attachment of cells to the matrix. Already 4 h post seeding, Phalloidin and DAPI staining visualized the interaction of SDSC with the surrounding matrix. When cultured for 7 days, a homogeneous distribution of viable cells became visible within the matrix (Fig 2A).

Bottom Line: High numbers of adult stem cells are still required to improve the formation of new vessels in scaffolds to accelerate dermal regeneration.Results showed that LPS-activated SDSC significantly enhanced vascularization of the scaffolds, compared to unstimulated stem cells in vivo.Our results suggest that combining activated stem cells and a dermal substitute is a promising option to enhance vascularization in scaffold-mediated dermal regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Plastic Surgery and Hand Surgery, University of Lübeck, Lübeck, Germany.

ABSTRACT
High numbers of adult stem cells are still required to improve the formation of new vessels in scaffolds to accelerate dermal regeneration. Recent data indicate a benefit for vascularization capacity by stimulating stem cells with lipopolysaccharide (LPS). In this study, stem cells derived from human skin (SDSC) were activated with LPS and seeded in a commercially available dermal substitute to examine vascularization in vivo. Besides, in vitro assays were performed to evaluate angiogenic factor release and tube formation ability. Results showed that LPS-activated SDSC significantly enhanced vascularization of the scaffolds, compared to unstimulated stem cells in vivo. Further, in vitro assays confirmed higher secretion rates of proangiogenic as well as proinflammatoric factors in the presence of LPS-activated SDSC. Our results suggest that combining activated stem cells and a dermal substitute is a promising option to enhance vascularization in scaffold-mediated dermal regeneration.

Show MeSH
Related in: MedlinePlus