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β2-adrenoceptor activation modulates skin wound healing processes to reduce scarring.

Le Provost GS, Pullar CE - J. Invest. Dermatol. (2014)

Bottom Line: Here we identify a β2AR-mediated mechanism for scar reduction. β2ARag significantly reduced HDF differentiation, via multiple cAMP and/or fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms, in the presence of transforming growth factor betaβ1, reduced contractile function, and inhibited mRNA expression of a number of profibrotic markers. β2ARag also reduced inflammation and angiogenesis in zebrafish and CAMs in vivo, respectively.In Red Duroc pig full-thickness wounds, β2ARag reduced both scar area and hyperpigmentation by almost 50% and significantly improved scar quality.Both macrophage infiltration and angiogenesis were initially decreased, whereas DF function was impaired in the β2ARag-treated porcine wound bed.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK.

ABSTRACT
During wound healing, excessive inflammation, angiogenesis, and differentiated human dermal fibroblast (HDF ) function contribute to scarring, whereas hyperpigmentation negatively affects scar quality. Over 100 million patients heal with a scar every year. To investigate the role of the beta 2 adrenergic receptor (β2AR) in wound scarring, the ability of beta 2 adrenergic receptor agonist (β2ARag) to alter HDF differentiation and function, wound inflammation, angiogenesis, and wound scarring was explored in HDFs, zebrafish, chick chorioallantoic membrane assay (CAM), and a porcine skin wound model, respectively. Here we identify a β2AR-mediated mechanism for scar reduction. β2ARag significantly reduced HDF differentiation, via multiple cAMP and/or fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms, in the presence of transforming growth factor betaβ1, reduced contractile function, and inhibited mRNA expression of a number of profibrotic markers. β2ARag also reduced inflammation and angiogenesis in zebrafish and CAMs in vivo, respectively. In Red Duroc pig full-thickness wounds, β2ARag reduced both scar area and hyperpigmentation by almost 50% and significantly improved scar quality. Indeed, mechanisms delineated in vitro and in other in vivo models were evident in the β2ARag-treated porcine scars in vivo. Both macrophage infiltration and angiogenesis were initially decreased, whereas DF function was impaired in the β2ARag-treated porcine wound bed. These data collectively reveal the potential of β2ARag to improve skin scarring.

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β2AR agonist (β2ARag) reduces the number of mature FAs on the human dermal fibroblast (HDF) periphery and profibrotic gene expression. HDFs were treated with serum-free medium (SFM) alone or containing β2ARag (Salbutamol, 10 μM), fibroblast growth factor 2 or basic FGF (FGF2) (10 ngml−1), PD173074 FGFR inhibitor (PD) (50 nM), sp-cAMP, rp-cAMP (50 μM), alone for 15 minutes or with pretreatment, as indicated. HDFs were pretreated with PD (a, b) or rp-cAMP (c) for 30 minutes, and then with β2ARag (a, c) or FGF2 (b) for a further 15 minutes. Mature (top) and supermature (bottom) FAs were measured and counted. Representative pictures are shown. Scale bar=10 μm. Data are presented as mean±SEM from at least 4 independent experiments (*P<0.05, **P<0.01). HDF gene expression was analyzed by RT-PCR, 6 hours post treatment with β2ARag (Salbutamol, Formoterol, 10 μM), in SFM. Mean mRNA levels were normalized to control values. Data are presented as mean±SEM from at least four independent experiments (*P<0.05, **P<0.01). CCN2, connective tissue growth factor; COL1A1, type 1 collagen, alpha 1; FGF2, fibroblast growth factor 2; FN EDA, fibronectin (FN) EDA; NS, not significant; SMA, smooth muscle α actin; TGFβ1, transforming growth factor beta 1.
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fig2: β2AR agonist (β2ARag) reduces the number of mature FAs on the human dermal fibroblast (HDF) periphery and profibrotic gene expression. HDFs were treated with serum-free medium (SFM) alone or containing β2ARag (Salbutamol, 10 μM), fibroblast growth factor 2 or basic FGF (FGF2) (10 ngml−1), PD173074 FGFR inhibitor (PD) (50 nM), sp-cAMP, rp-cAMP (50 μM), alone for 15 minutes or with pretreatment, as indicated. HDFs were pretreated with PD (a, b) or rp-cAMP (c) for 30 minutes, and then with β2ARag (a, c) or FGF2 (b) for a further 15 minutes. Mature (top) and supermature (bottom) FAs were measured and counted. Representative pictures are shown. Scale bar=10 μm. Data are presented as mean±SEM from at least 4 independent experiments (*P<0.05, **P<0.01). HDF gene expression was analyzed by RT-PCR, 6 hours post treatment with β2ARag (Salbutamol, Formoterol, 10 μM), in SFM. Mean mRNA levels were normalized to control values. Data are presented as mean±SEM from at least four independent experiments (*P<0.05, **P<0.01). CCN2, connective tissue growth factor; COL1A1, type 1 collagen, alpha 1; FGF2, fibroblast growth factor 2; FN EDA, fibronectin (FN) EDA; NS, not significant; SMA, smooth muscle α actin; TGFβ1, transforming growth factor beta 1.

Mentions: To determine whether β2ARag altered the number of M/supermature focal adhesion (SMFAs), the length of FAs at the HDF periphery was analyzed. β2ARag reduced both peripheral HDF MFA and SMFA numbers by 23 and 58%, respectively (Figure 2a). PD was used to probe FGF2 involvement, and it had no effect alone and it did not prevent the β2ARag-mediated reduction in M/SMFA numbers (Figure 2a). However, exogenous FGF2 also reduced HDF peripheral MFA and SMFA numbers by 29 and 63%, respectively, whereas PD completely prevented any decrease (Figure 2b).


β2-adrenoceptor activation modulates skin wound healing processes to reduce scarring.

Le Provost GS, Pullar CE - J. Invest. Dermatol. (2014)

β2AR agonist (β2ARag) reduces the number of mature FAs on the human dermal fibroblast (HDF) periphery and profibrotic gene expression. HDFs were treated with serum-free medium (SFM) alone or containing β2ARag (Salbutamol, 10 μM), fibroblast growth factor 2 or basic FGF (FGF2) (10 ngml−1), PD173074 FGFR inhibitor (PD) (50 nM), sp-cAMP, rp-cAMP (50 μM), alone for 15 minutes or with pretreatment, as indicated. HDFs were pretreated with PD (a, b) or rp-cAMP (c) for 30 minutes, and then with β2ARag (a, c) or FGF2 (b) for a further 15 minutes. Mature (top) and supermature (bottom) FAs were measured and counted. Representative pictures are shown. Scale bar=10 μm. Data are presented as mean±SEM from at least 4 independent experiments (*P<0.05, **P<0.01). HDF gene expression was analyzed by RT-PCR, 6 hours post treatment with β2ARag (Salbutamol, Formoterol, 10 μM), in SFM. Mean mRNA levels were normalized to control values. Data are presented as mean±SEM from at least four independent experiments (*P<0.05, **P<0.01). CCN2, connective tissue growth factor; COL1A1, type 1 collagen, alpha 1; FGF2, fibroblast growth factor 2; FN EDA, fibronectin (FN) EDA; NS, not significant; SMA, smooth muscle α actin; TGFβ1, transforming growth factor beta 1.
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fig2: β2AR agonist (β2ARag) reduces the number of mature FAs on the human dermal fibroblast (HDF) periphery and profibrotic gene expression. HDFs were treated with serum-free medium (SFM) alone or containing β2ARag (Salbutamol, 10 μM), fibroblast growth factor 2 or basic FGF (FGF2) (10 ngml−1), PD173074 FGFR inhibitor (PD) (50 nM), sp-cAMP, rp-cAMP (50 μM), alone for 15 minutes or with pretreatment, as indicated. HDFs were pretreated with PD (a, b) or rp-cAMP (c) for 30 minutes, and then with β2ARag (a, c) or FGF2 (b) for a further 15 minutes. Mature (top) and supermature (bottom) FAs were measured and counted. Representative pictures are shown. Scale bar=10 μm. Data are presented as mean±SEM from at least 4 independent experiments (*P<0.05, **P<0.01). HDF gene expression was analyzed by RT-PCR, 6 hours post treatment with β2ARag (Salbutamol, Formoterol, 10 μM), in SFM. Mean mRNA levels were normalized to control values. Data are presented as mean±SEM from at least four independent experiments (*P<0.05, **P<0.01). CCN2, connective tissue growth factor; COL1A1, type 1 collagen, alpha 1; FGF2, fibroblast growth factor 2; FN EDA, fibronectin (FN) EDA; NS, not significant; SMA, smooth muscle α actin; TGFβ1, transforming growth factor beta 1.
Mentions: To determine whether β2ARag altered the number of M/supermature focal adhesion (SMFAs), the length of FAs at the HDF periphery was analyzed. β2ARag reduced both peripheral HDF MFA and SMFA numbers by 23 and 58%, respectively (Figure 2a). PD was used to probe FGF2 involvement, and it had no effect alone and it did not prevent the β2ARag-mediated reduction in M/SMFA numbers (Figure 2a). However, exogenous FGF2 also reduced HDF peripheral MFA and SMFA numbers by 29 and 63%, respectively, whereas PD completely prevented any decrease (Figure 2b).

Bottom Line: Here we identify a β2AR-mediated mechanism for scar reduction. β2ARag significantly reduced HDF differentiation, via multiple cAMP and/or fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms, in the presence of transforming growth factor betaβ1, reduced contractile function, and inhibited mRNA expression of a number of profibrotic markers. β2ARag also reduced inflammation and angiogenesis in zebrafish and CAMs in vivo, respectively.In Red Duroc pig full-thickness wounds, β2ARag reduced both scar area and hyperpigmentation by almost 50% and significantly improved scar quality.Both macrophage infiltration and angiogenesis were initially decreased, whereas DF function was impaired in the β2ARag-treated porcine wound bed.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK.

ABSTRACT
During wound healing, excessive inflammation, angiogenesis, and differentiated human dermal fibroblast (HDF ) function contribute to scarring, whereas hyperpigmentation negatively affects scar quality. Over 100 million patients heal with a scar every year. To investigate the role of the beta 2 adrenergic receptor (β2AR) in wound scarring, the ability of beta 2 adrenergic receptor agonist (β2ARag) to alter HDF differentiation and function, wound inflammation, angiogenesis, and wound scarring was explored in HDFs, zebrafish, chick chorioallantoic membrane assay (CAM), and a porcine skin wound model, respectively. Here we identify a β2AR-mediated mechanism for scar reduction. β2ARag significantly reduced HDF differentiation, via multiple cAMP and/or fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms, in the presence of transforming growth factor betaβ1, reduced contractile function, and inhibited mRNA expression of a number of profibrotic markers. β2ARag also reduced inflammation and angiogenesis in zebrafish and CAMs in vivo, respectively. In Red Duroc pig full-thickness wounds, β2ARag reduced both scar area and hyperpigmentation by almost 50% and significantly improved scar quality. Indeed, mechanisms delineated in vitro and in other in vivo models were evident in the β2ARag-treated porcine scars in vivo. Both macrophage infiltration and angiogenesis were initially decreased, whereas DF function was impaired in the β2ARag-treated porcine wound bed. These data collectively reveal the potential of β2ARag to improve skin scarring.

Show MeSH
Related in: MedlinePlus