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Hyaluronidase modulates inflammatory response and accelerates the cutaneous wound healing.

Fronza M, Caetano GF, Leite MN, Bitencourt CS, Paula-Silva FW, Andrade TA, Frade MA, Merfort I, Faccioli LH - PLoS ONE (2014)

Bottom Line: Hyaluronidases are enzymes that degrade hyaluronan an important constituent of the extracellular matrix.They have been used as a spreading agent, improving the absorption of drugs and facilitating the subcutaneous infusion of fluids.Moreover, HYAL increased gene expression of peroxisome proliferator-activated receptors (PPAR) γ and PPAR β/δ, the collagen content in the early stages of healing processes as well as angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil; Departamento de Farmácia, Universidade de Vila Velha, Vila Velha, Espirito Santo, Brazil.

ABSTRACT
Hyaluronidases are enzymes that degrade hyaluronan an important constituent of the extracellular matrix. They have been used as a spreading agent, improving the absorption of drugs and facilitating the subcutaneous infusion of fluids. Here, we investigated the influence of bovine testes hyaluronidase (HYAL) during cutaneous wound healing in in vitro and in vivo assays. We demonstrated in the wound scratch assay that HYAL increased the migration and proliferation of fibroblasts in vitro at low concentration, e.g. 0.1 U HYAL enhanced the cell number by 20%. HYAL presented faster and higher reepithelialization in in vivo full-thickness excisional wounds generated on adult Wistar rats back skin already in the early phase at 2nd day post operatory compared to vehicle-control group. Wound closured area observed in the 16 U and 32 U HYAL treated rats reached 38% and 46% compared to 19% in the controls, respectively. Histological and biochemical analyses supported the clinical observations and showed that HYAL treated wounds exhibited increased granulation tissue, diminished edema formation and regulated the inflammatory response by modulating the release of pro and anti-inflammatory cytokines, growth factor and eicosanoids mediators. Moreover, HYAL increased gene expression of peroxisome proliferator-activated receptors (PPAR) γ and PPAR β/δ, the collagen content in the early stages of healing processes as well as angiogenesis. Altogether these data revealed that HYAL accelerates wound healing processes and might be beneficial for treating wound disorders.

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Neovascularization induced by HYAL.Animals were topically treated either with vehicle (control group) or HYAL 16 U daily. Paraffin-wound sections were stained with HE to evaluate the angiogenic response by image analysis. The sections were photographed at 400x. The ImageJ software was used to count the blood vessels in wound tissue specimens at day 2, 7, 14 and 21 post wounding in at least ten random optic fields per group. (A) Representative photomicrograph of blood vessels (black arrow) in wound tissue specimens at day 21 post-wounding. (B) Histogram of a quantitative analysis of vascular density counted. (C) Vascular endothelial growth factor (VEGF) expression measured in the supernatant of wound tissue homogenate by ELISA. Data represent means ± SEM (n = 8 wounds/group), **P<0.01, ***P<0.001 compared to control group by one-way-ANOVA.
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pone-0112297-g005: Neovascularization induced by HYAL.Animals were topically treated either with vehicle (control group) or HYAL 16 U daily. Paraffin-wound sections were stained with HE to evaluate the angiogenic response by image analysis. The sections were photographed at 400x. The ImageJ software was used to count the blood vessels in wound tissue specimens at day 2, 7, 14 and 21 post wounding in at least ten random optic fields per group. (A) Representative photomicrograph of blood vessels (black arrow) in wound tissue specimens at day 21 post-wounding. (B) Histogram of a quantitative analysis of vascular density counted. (C) Vascular endothelial growth factor (VEGF) expression measured in the supernatant of wound tissue homogenate by ELISA. Data represent means ± SEM (n = 8 wounds/group), **P<0.01, ***P<0.001 compared to control group by one-way-ANOVA.

Mentions: The number of blood vessels in the HYAL treated wound was determined using different techniques. Figure 5A exemplarily illustrates photomicrographs of blood vessels in tissue sections from the wounds. The number of blood vessels in the 16 U HYAL treated wounds determined by morphometric analyses was increased after 7, 14 and 21 days (Figure 5B). Levels of VEGF, a signal protein that stimulates angiogenesis, measured by ELISA in the wound tissue homogenate was enhanced in 16 U HYAL treated wounds at day 2 post wounding (Figure 5C).


Hyaluronidase modulates inflammatory response and accelerates the cutaneous wound healing.

Fronza M, Caetano GF, Leite MN, Bitencourt CS, Paula-Silva FW, Andrade TA, Frade MA, Merfort I, Faccioli LH - PLoS ONE (2014)

Neovascularization induced by HYAL.Animals were topically treated either with vehicle (control group) or HYAL 16 U daily. Paraffin-wound sections were stained with HE to evaluate the angiogenic response by image analysis. The sections were photographed at 400x. The ImageJ software was used to count the blood vessels in wound tissue specimens at day 2, 7, 14 and 21 post wounding in at least ten random optic fields per group. (A) Representative photomicrograph of blood vessels (black arrow) in wound tissue specimens at day 21 post-wounding. (B) Histogram of a quantitative analysis of vascular density counted. (C) Vascular endothelial growth factor (VEGF) expression measured in the supernatant of wound tissue homogenate by ELISA. Data represent means ± SEM (n = 8 wounds/group), **P<0.01, ***P<0.001 compared to control group by one-way-ANOVA.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230982&req=5

pone-0112297-g005: Neovascularization induced by HYAL.Animals were topically treated either with vehicle (control group) or HYAL 16 U daily. Paraffin-wound sections were stained with HE to evaluate the angiogenic response by image analysis. The sections were photographed at 400x. The ImageJ software was used to count the blood vessels in wound tissue specimens at day 2, 7, 14 and 21 post wounding in at least ten random optic fields per group. (A) Representative photomicrograph of blood vessels (black arrow) in wound tissue specimens at day 21 post-wounding. (B) Histogram of a quantitative analysis of vascular density counted. (C) Vascular endothelial growth factor (VEGF) expression measured in the supernatant of wound tissue homogenate by ELISA. Data represent means ± SEM (n = 8 wounds/group), **P<0.01, ***P<0.001 compared to control group by one-way-ANOVA.
Mentions: The number of blood vessels in the HYAL treated wound was determined using different techniques. Figure 5A exemplarily illustrates photomicrographs of blood vessels in tissue sections from the wounds. The number of blood vessels in the 16 U HYAL treated wounds determined by morphometric analyses was increased after 7, 14 and 21 days (Figure 5B). Levels of VEGF, a signal protein that stimulates angiogenesis, measured by ELISA in the wound tissue homogenate was enhanced in 16 U HYAL treated wounds at day 2 post wounding (Figure 5C).

Bottom Line: Hyaluronidases are enzymes that degrade hyaluronan an important constituent of the extracellular matrix.They have been used as a spreading agent, improving the absorption of drugs and facilitating the subcutaneous infusion of fluids.Moreover, HYAL increased gene expression of peroxisome proliferator-activated receptors (PPAR) γ and PPAR β/δ, the collagen content in the early stages of healing processes as well as angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil; Departamento de Farmácia, Universidade de Vila Velha, Vila Velha, Espirito Santo, Brazil.

ABSTRACT
Hyaluronidases are enzymes that degrade hyaluronan an important constituent of the extracellular matrix. They have been used as a spreading agent, improving the absorption of drugs and facilitating the subcutaneous infusion of fluids. Here, we investigated the influence of bovine testes hyaluronidase (HYAL) during cutaneous wound healing in in vitro and in vivo assays. We demonstrated in the wound scratch assay that HYAL increased the migration and proliferation of fibroblasts in vitro at low concentration, e.g. 0.1 U HYAL enhanced the cell number by 20%. HYAL presented faster and higher reepithelialization in in vivo full-thickness excisional wounds generated on adult Wistar rats back skin already in the early phase at 2nd day post operatory compared to vehicle-control group. Wound closured area observed in the 16 U and 32 U HYAL treated rats reached 38% and 46% compared to 19% in the controls, respectively. Histological and biochemical analyses supported the clinical observations and showed that HYAL treated wounds exhibited increased granulation tissue, diminished edema formation and regulated the inflammatory response by modulating the release of pro and anti-inflammatory cytokines, growth factor and eicosanoids mediators. Moreover, HYAL increased gene expression of peroxisome proliferator-activated receptors (PPAR) γ and PPAR β/δ, the collagen content in the early stages of healing processes as well as angiogenesis. Altogether these data revealed that HYAL accelerates wound healing processes and might be beneficial for treating wound disorders.

Show MeSH
Related in: MedlinePlus