Limits...
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.

Show MeSH

Related in: MedlinePlus

Topical application of HYAL accelerates wound closure in full-thickness excisional wounds.(A) Representative photographs taken from the 150 mm diameter full-thickness wounds of the Wistar rats. Macroscopic changes in skin wound sites induced by topical application of vehicle, HYAL 16 U and HYAL 32 U at day 0 (picture taken immediately after injury) 2, 5, 7, 10, 14 and day 21 are shown. (B) Rate of wound closure induced by topical application of vehicle (control group), HYAL 16 U and HYAL 32 U at day 2, 5, 7, 10, 14 and day 21 are given. Data are expressed as percentage of reduction area from the original wound size (day zero). Values are mean ± SEM (n = 8 to 16 wounds/group), *P<0.05, **P<0.01, ***P<0.001 compared to control group by two-way-ANOVA.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4230982&req=5

pone-0112297-g001: Topical application of HYAL accelerates wound closure in full-thickness excisional wounds.(A) Representative photographs taken from the 150 mm diameter full-thickness wounds of the Wistar rats. Macroscopic changes in skin wound sites induced by topical application of vehicle, HYAL 16 U and HYAL 32 U at day 0 (picture taken immediately after injury) 2, 5, 7, 10, 14 and day 21 are shown. (B) Rate of wound closure induced by topical application of vehicle (control group), HYAL 16 U and HYAL 32 U at day 2, 5, 7, 10, 14 and day 21 are given. Data are expressed as percentage of reduction area from the original wound size (day zero). Values are mean ± SEM (n = 8 to 16 wounds/group), *P<0.05, **P<0.01, ***P<0.001 compared to control group by two-way-ANOVA.

Mentions: As showed in Figure 1, the wounds treated with 16 U or 32 U HYAL presented faster and higher reepithelialization compared to vehicle-control group. Selected doses of 16 U and 32 U were based on the in vitro scratch assay, in which these concentrations exhibited the maximum stimulatory effects on fibroblasts proliferation and migration, as well as on our previous in vivo studies using HYAL [20]. Wound closure area observed in the 16 U and 32 U HYAL reached 38% and 46% compared to only 19% in the controls already in the early stage (2nd day), respectively. After 5 and 7 days the closure wound in the 16 U and 32 U HYAL reached 67% and 68% (at day 5) and 85% and 89% (at day 7) as compare to 40% and 69% observed in control group, respectively. Interesting, by day 14 the wounds of HYAL-treated rats were completely closed, whereas the wounds of control rats had not yet completely healed. This observation suggests that HYAL positively influenced the wound healing process and that 16 U or 32 U HYAL treatments healed the full thickness wound in a similar speed. Therefore, the following experiments were performed using only 16 U HYAL.


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)

Topical application of HYAL accelerates wound closure in full-thickness excisional wounds.(A) Representative photographs taken from the 150 mm diameter full-thickness wounds of the Wistar rats. Macroscopic changes in skin wound sites induced by topical application of vehicle, HYAL 16 U and HYAL 32 U at day 0 (picture taken immediately after injury) 2, 5, 7, 10, 14 and day 21 are shown. (B) Rate of wound closure induced by topical application of vehicle (control group), HYAL 16 U and HYAL 32 U at day 2, 5, 7, 10, 14 and day 21 are given. Data are expressed as percentage of reduction area from the original wound size (day zero). Values are mean ± SEM (n = 8 to 16 wounds/group), *P<0.05, **P<0.01, ***P<0.001 compared to control group by two-way-ANOVA.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0112297-g001: Topical application of HYAL accelerates wound closure in full-thickness excisional wounds.(A) Representative photographs taken from the 150 mm diameter full-thickness wounds of the Wistar rats. Macroscopic changes in skin wound sites induced by topical application of vehicle, HYAL 16 U and HYAL 32 U at day 0 (picture taken immediately after injury) 2, 5, 7, 10, 14 and day 21 are shown. (B) Rate of wound closure induced by topical application of vehicle (control group), HYAL 16 U and HYAL 32 U at day 2, 5, 7, 10, 14 and day 21 are given. Data are expressed as percentage of reduction area from the original wound size (day zero). Values are mean ± SEM (n = 8 to 16 wounds/group), *P<0.05, **P<0.01, ***P<0.001 compared to control group by two-way-ANOVA.
Mentions: As showed in Figure 1, the wounds treated with 16 U or 32 U HYAL presented faster and higher reepithelialization compared to vehicle-control group. Selected doses of 16 U and 32 U were based on the in vitro scratch assay, in which these concentrations exhibited the maximum stimulatory effects on fibroblasts proliferation and migration, as well as on our previous in vivo studies using HYAL [20]. Wound closure area observed in the 16 U and 32 U HYAL reached 38% and 46% compared to only 19% in the controls already in the early stage (2nd day), respectively. After 5 and 7 days the closure wound in the 16 U and 32 U HYAL reached 67% and 68% (at day 5) and 85% and 89% (at day 7) as compare to 40% and 69% observed in control group, respectively. Interesting, by day 14 the wounds of HYAL-treated rats were completely closed, whereas the wounds of control rats had not yet completely healed. This observation suggests that HYAL positively influenced the wound healing process and that 16 U or 32 U HYAL treatments healed the full thickness wound in a similar speed. Therefore, the following experiments were performed using only 16 U HYAL.

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