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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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PPARs gene expression in skin biopsies dissected from the cutaneous wounds after HYAL treatment analyzed by qRT-PCR.A) mRNA for PPAR α, B) PPAR δ and C) PPAR γ determined by qRT-PCR. Ppar expression was normalized by Gapdh and beta-actin. Animals were topically treated with the vehicle (control group) or 16 U HYAL daily for 2, 7, 14 and 21 days, respectively. 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-g006: PPARs gene expression in skin biopsies dissected from the cutaneous wounds after HYAL treatment analyzed by qRT-PCR.A) mRNA for PPAR α, B) PPAR δ and C) PPAR γ determined by qRT-PCR. Ppar expression was normalized by Gapdh and beta-actin. Animals were topically treated with the vehicle (control group) or 16 U HYAL daily for 2, 7, 14 and 21 days, respectively. Data represent means ± SEM (n = 8 wounds/group), **P<0.01, ***P<0.001 compared to control group by one-way-ANOVA.

Mentions: PPAR α and δ (also called β) have been shown to be important for the rapid epithelialization of wound skin and PPAR γ during the resolution phase of wound repair. To study the impact of HYAL on PPAR gene expression (Ppara, Pparg and Ppard), the skin extracts dissected from the cutaneous wounds were analyzed by qRT-PCR. The three PPAR isotypes were expressed in the skin at all-time points, but in a different manner (Figure 6). mRNA for PPAR α (Ppara) was inhibited in the cutaneous wound, compared to unwounded skin (day 0). Interestingly, a time-dependent increased expression of Ppara was observed after daily treatment of 16 U HYAL until day 21, reaching finally a similar expression level as the physiological one (Figure 6A). Levels of PPAR δ mRNA were only increased at day 2 (fold increase 1.62), but returned to levels measured in the unwounded tissue (Figure 6B). mRNA of PPAR γ showed the highest fold increase at day 2 (fold increase 5.3) and decreased to levels in the range of the controls (Figure 6C).


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)

PPARs gene expression in skin biopsies dissected from the cutaneous wounds after HYAL treatment analyzed by qRT-PCR.A) mRNA for PPAR α, B) PPAR δ and C) PPAR γ determined by qRT-PCR. Ppar expression was normalized by Gapdh and beta-actin. Animals were topically treated with the vehicle (control group) or 16 U HYAL daily for 2, 7, 14 and 21 days, respectively. 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-g006: PPARs gene expression in skin biopsies dissected from the cutaneous wounds after HYAL treatment analyzed by qRT-PCR.A) mRNA for PPAR α, B) PPAR δ and C) PPAR γ determined by qRT-PCR. Ppar expression was normalized by Gapdh and beta-actin. Animals were topically treated with the vehicle (control group) or 16 U HYAL daily for 2, 7, 14 and 21 days, respectively. Data represent means ± SEM (n = 8 wounds/group), **P<0.01, ***P<0.001 compared to control group by one-way-ANOVA.
Mentions: PPAR α and δ (also called β) have been shown to be important for the rapid epithelialization of wound skin and PPAR γ during the resolution phase of wound repair. To study the impact of HYAL on PPAR gene expression (Ppara, Pparg and Ppard), the skin extracts dissected from the cutaneous wounds were analyzed by qRT-PCR. The three PPAR isotypes were expressed in the skin at all-time points, but in a different manner (Figure 6). mRNA for PPAR α (Ppara) was inhibited in the cutaneous wound, compared to unwounded skin (day 0). Interestingly, a time-dependent increased expression of Ppara was observed after daily treatment of 16 U HYAL until day 21, reaching finally a similar expression level as the physiological one (Figure 6A). Levels of PPAR δ mRNA were only increased at day 2 (fold increase 1.62), but returned to levels measured in the unwounded tissue (Figure 6B). mRNA of PPAR γ showed the highest fold increase at day 2 (fold increase 5.3) and decreased to levels in the range of the controls (Figure 6C).

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