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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 human dermal fibroblast (HDF) differentiation and contractile function in vitro via cAMP- and fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms. HDFs were treated with serum-free medium (SFM) alone or SFM containing Salbutamol/Formoterol (10 μM), TGFβ1 (1 ng ml−1), FGF2 (10 ng ml−1), PD173074 FGFR inhibitor (PD173074,50 nM), sp-cAMP/rp-cAMP (50 μM), alone or pretreated, as indicated. (a) HDFs were treated for 48 hours with TGFβ1 alone or 6 hours pretreatment with TGFβ1 or β2ARag before β2ARag or TGFβ1 addition, respectively, for 42 hours. Ratios of smooth muscle α actin (SMA)-positive cells to total cells were normalized to control. Scale bar=100 μm. (b) HDF FGF2 secretion was ELISA analyzed 6/24 hours post treatment in the presence or absence of β2ARag. (c) HDFs were treated for 48 hours with FGF2, PD, or β2ARag, with/without 6 hours of PD pretreatment. (d) HDFs were treated for 2 hours with sp-cAMP, rp-cAMP, or β2ARag, with/without rp-cAMP pretreatment, 30 minutes before β2ARag for 2 hours. Data presented are means±SEM; 4 independent experiments. NS, not significant.
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fig1: β2AR agonist (β2ARag) reduces human dermal fibroblast (HDF) differentiation and contractile function in vitro via cAMP- and fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms. HDFs were treated with serum-free medium (SFM) alone or SFM containing Salbutamol/Formoterol (10 μM), TGFβ1 (1 ng ml−1), FGF2 (10 ng ml−1), PD173074 FGFR inhibitor (PD173074,50 nM), sp-cAMP/rp-cAMP (50 μM), alone or pretreated, as indicated. (a) HDFs were treated for 48 hours with TGFβ1 alone or 6 hours pretreatment with TGFβ1 or β2ARag before β2ARag or TGFβ1 addition, respectively, for 42 hours. Ratios of smooth muscle α actin (SMA)-positive cells to total cells were normalized to control. Scale bar=100 μm. (b) HDF FGF2 secretion was ELISA analyzed 6/24 hours post treatment in the presence or absence of β2ARag. (c) HDFs were treated for 48 hours with FGF2, PD, or β2ARag, with/without 6 hours of PD pretreatment. (d) HDFs were treated for 2 hours with sp-cAMP, rp-cAMP, or β2ARag, with/without rp-cAMP pretreatment, 30 minutes before β2ARag for 2 hours. Data presented are means±SEM; 4 independent experiments. NS, not significant.

Mentions: Transforming growth factor beta (TGFβ)1, a fibroblast differentiation promoter strongly upregulated in wounds (Hinz, 2007), increased the ratio of SMA-positive HDFs by 14.5-fold (Figure 1a). Regardless of whether the β2ARag was added 6 hours prior or subsequent to TGFβ1 for a further 42 hours, similar, marked inhibition was observed, decreasing the ratio of SMA-positive HDFs by up to 95% (Figure 1a).


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

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

β2AR agonist (β2ARag) reduces human dermal fibroblast (HDF) differentiation and contractile function in vitro via cAMP- and fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms. HDFs were treated with serum-free medium (SFM) alone or SFM containing Salbutamol/Formoterol (10 μM), TGFβ1 (1 ng ml−1), FGF2 (10 ng ml−1), PD173074 FGFR inhibitor (PD173074,50 nM), sp-cAMP/rp-cAMP (50 μM), alone or pretreated, as indicated. (a) HDFs were treated for 48 hours with TGFβ1 alone or 6 hours pretreatment with TGFβ1 or β2ARag before β2ARag or TGFβ1 addition, respectively, for 42 hours. Ratios of smooth muscle α actin (SMA)-positive cells to total cells were normalized to control. Scale bar=100 μm. (b) HDF FGF2 secretion was ELISA analyzed 6/24 hours post treatment in the presence or absence of β2ARag. (c) HDFs were treated for 48 hours with FGF2, PD, or β2ARag, with/without 6 hours of PD pretreatment. (d) HDFs were treated for 2 hours with sp-cAMP, rp-cAMP, or β2ARag, with/without rp-cAMP pretreatment, 30 minutes before β2ARag for 2 hours. Data presented are means±SEM; 4 independent experiments. NS, not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4263603&req=5

fig1: β2AR agonist (β2ARag) reduces human dermal fibroblast (HDF) differentiation and contractile function in vitro via cAMP- and fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms. HDFs were treated with serum-free medium (SFM) alone or SFM containing Salbutamol/Formoterol (10 μM), TGFβ1 (1 ng ml−1), FGF2 (10 ng ml−1), PD173074 FGFR inhibitor (PD173074,50 nM), sp-cAMP/rp-cAMP (50 μM), alone or pretreated, as indicated. (a) HDFs were treated for 48 hours with TGFβ1 alone or 6 hours pretreatment with TGFβ1 or β2ARag before β2ARag or TGFβ1 addition, respectively, for 42 hours. Ratios of smooth muscle α actin (SMA)-positive cells to total cells were normalized to control. Scale bar=100 μm. (b) HDF FGF2 secretion was ELISA analyzed 6/24 hours post treatment in the presence or absence of β2ARag. (c) HDFs were treated for 48 hours with FGF2, PD, or β2ARag, with/without 6 hours of PD pretreatment. (d) HDFs were treated for 2 hours with sp-cAMP, rp-cAMP, or β2ARag, with/without rp-cAMP pretreatment, 30 minutes before β2ARag for 2 hours. Data presented are means±SEM; 4 independent experiments. NS, not significant.
Mentions: Transforming growth factor beta (TGFβ)1, a fibroblast differentiation promoter strongly upregulated in wounds (Hinz, 2007), increased the ratio of SMA-positive HDFs by 14.5-fold (Figure 1a). Regardless of whether the β2ARag was added 6 hours prior or subsequent to TGFβ1 for a further 42 hours, similar, marked inhibition was observed, decreasing the ratio of SMA-positive HDFs by up to 95% (Figure 1a).

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