Effects of topical copper tripeptide complex on wound healing in an irradiated rat model

Noah P. Parker; Farhad Ardeshirpour; Stephen C. Schmechel; Amy Anne D Lassig

doi:10.1177/0194599813492644

Effects of topical copper tripeptide complex on wound healing in an irradiated rat model

Noah P. Parker, Farhad Ardeshirpour, Stephen C. Schmechel, Amy Anne D Lassig

Otolaryngology

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Objectives. To evaluate clinical and immunohistopathological effects of topical glycyl-histidyl-lysine-copper (GHK-Cu) on in vivo irradiated rat wounds. Design. Animal model. Setting. Academic institution. Subjects and Methods. After dorsal irradiation and a 28-day recovery period, 2 × 8 cm cranially based dorsal flaps were created in Sprague-Dawley rats. Twice daily GHK-Cu gel (test) or aquaphilic ointment (control) was applied for 10 days. Animals were euthanized, digital images of flaps were taken, and harvested tissues were immunohistochemically stained for a vascular endothelium marker, caveolin-1, and vascular endothelial growth factor (VEGF). Digital image analysis was used for outcome measures. Unpaired t-tests were used for statistical analyses; significance of P < .01 accounted for multiple comparisons. Results. By digital analysis of clinical images, 13 test and 10 control animals showed mean ischemic areas of 5.0 cm² (SD = 0.9) for tests and 3.8 cm² (SD = 1.1; P = .011) for controls. Whole slide digitized images allowed quantification of caveolin-1-stained blood vessels and VEGF expression in fibroblasts at the interface of healing flaps. Caveolin-1 analyses showed a mean of 209.0 vessels (SD = 111.1) and a mean vessel luminal area of 525.7 um ² (SD = 191.0) in tests and 207.4 vessels (SD = 109.4; P = .973) and 422.8 um² (SD = 109.7; P = .118) in controls. VEGF quantified as the percentage of pixels exceeding a colorimetric threshold, with higher fractions of positive pixels indicating more intense staining, showed a mean intensity score of 0.34 (SD = 0.19) in tests and 0.54 (SD = 0.41; P = .169) in controls. Conclusions. Irradiated dorsal rat flaps treated with topical GHK-Cu gel demonstrated no difference in flap ischemia, blood vessel number or area, or VEGF expression compared to controls.

Original language	English (US)
Pages (from-to)	384-389
Number of pages	6
Journal	Otolaryngology - Head and Neck Surgery (United States)
Volume	149
Issue number	3
DOIs	https://doi.org/10.1177/0194599813492644
State	Published - Sep 2013

Bibliographical note

Funding Information:
Funding source: 5M Lions Club Research Grant and NIH grants P30-CA77598 (D Yee), P50-CA101955 (D Buchsbaum), KL2-RR033182 (B Blazar), NCI Contract No. HHSN261200800001E (S Schmechel), funding purchasing of animals, conducting the live animal portions of the study, purchase of appropriate immunohistochemical staining, and conducting the histological and immunohistochemical portion of the study. Funding the digital imaging and analysis of immunohistochemically stained slides.

Funding Information:
This project utilized BioNet core facilities which are supported by the University of Minnesota Academic Health Center. Specifically, Colleen Forster from the BioNet histology core provided expertise in histology and immunohistochemistry, and Jonathan Henriksen of the BioNet digital imaging core provided expertise in digital image analysis. We are indebted to Kim Janisch, Susan Jenson, and Dr Dan Feeney, DVM, from the University of Minnesota Veterinary Medical Center for their assistance with animal irradiation.

Keywords

animal model
copper tripeptide
irradiation
radiation
rat
wound healing

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title = "Effects of topical copper tripeptide complex on wound healing in an irradiated rat model",

abstract = "Objectives. To evaluate clinical and immunohistopathological effects of topical glycyl-histidyl-lysine-copper (GHK-Cu) on in vivo irradiated rat wounds. Design. Animal model. Setting. Academic institution. Subjects and Methods. After dorsal irradiation and a 28-day recovery period, 2 × 8 cm cranially based dorsal flaps were created in Sprague-Dawley rats. Twice daily GHK-Cu gel (test) or aquaphilic ointment (control) was applied for 10 days. Animals were euthanized, digital images of flaps were taken, and harvested tissues were immunohistochemically stained for a vascular endothelium marker, caveolin-1, and vascular endothelial growth factor (VEGF). Digital image analysis was used for outcome measures. Unpaired t-tests were used for statistical analyses; significance of P < .01 accounted for multiple comparisons. Results. By digital analysis of clinical images, 13 test and 10 control animals showed mean ischemic areas of 5.0 cm2 (SD = 0.9) for tests and 3.8 cm2 (SD = 1.1; P = .011) for controls. Whole slide digitized images allowed quantification of caveolin-1-stained blood vessels and VEGF expression in fibroblasts at the interface of healing flaps. Caveolin-1 analyses showed a mean of 209.0 vessels (SD = 111.1) and a mean vessel luminal area of 525.7 um 2 (SD = 191.0) in tests and 207.4 vessels (SD = 109.4; P = .973) and 422.8 um2 (SD = 109.7; P = .118) in controls. VEGF quantified as the percentage of pixels exceeding a colorimetric threshold, with higher fractions of positive pixels indicating more intense staining, showed a mean intensity score of 0.34 (SD = 0.19) in tests and 0.54 (SD = 0.41; P = .169) in controls. Conclusions. Irradiated dorsal rat flaps treated with topical GHK-Cu gel demonstrated no difference in flap ischemia, blood vessel number or area, or VEGF expression compared to controls.",

keywords = "animal model, copper tripeptide, irradiation, radiation, rat, wound healing",

author = "Parker, {Noah P.} and Farhad Ardeshirpour and Schmechel, {Stephen C.} and Lassig, {Amy Anne D}",

note = "Funding Information: Funding source: 5M Lions Club Research Grant and NIH grants P30-CA77598 (D Yee), P50-CA101955 (D Buchsbaum), KL2-RR033182 (B Blazar), NCI Contract No. HHSN261200800001E (S Schmechel), funding purchasing of animals, conducting the live animal portions of the study, purchase of appropriate immunohistochemical staining, and conducting the histological and immunohistochemical portion of the study. Funding the digital imaging and analysis of immunohistochemically stained slides. Funding Information: This project utilized BioNet core facilities which are supported by the University of Minnesota Academic Health Center. Specifically, Colleen Forster from the BioNet histology core provided expertise in histology and immunohistochemistry, and Jonathan Henriksen of the BioNet digital imaging core provided expertise in digital image analysis. We are indebted to Kim Janisch, Susan Jenson, and Dr Dan Feeney, DVM, from the University of Minnesota Veterinary Medical Center for their assistance with animal irradiation. ",

year = "2013",

month = sep,

doi = "10.1177/0194599813492644",

language = "English (US)",

volume = "149",

pages = "384--389",

journal = "Otolaryngology - Head and Neck Surgery (United States)",

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number = "3",

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TY - JOUR

T1 - Effects of topical copper tripeptide complex on wound healing in an irradiated rat model

AU - Parker, Noah P.

AU - Ardeshirpour, Farhad

AU - Schmechel, Stephen C.

AU - Lassig, Amy Anne D

N1 - Funding Information: Funding source: 5M Lions Club Research Grant and NIH grants P30-CA77598 (D Yee), P50-CA101955 (D Buchsbaum), KL2-RR033182 (B Blazar), NCI Contract No. HHSN261200800001E (S Schmechel), funding purchasing of animals, conducting the live animal portions of the study, purchase of appropriate immunohistochemical staining, and conducting the histological and immunohistochemical portion of the study. Funding the digital imaging and analysis of immunohistochemically stained slides. Funding Information: This project utilized BioNet core facilities which are supported by the University of Minnesota Academic Health Center. Specifically, Colleen Forster from the BioNet histology core provided expertise in histology and immunohistochemistry, and Jonathan Henriksen of the BioNet digital imaging core provided expertise in digital image analysis. We are indebted to Kim Janisch, Susan Jenson, and Dr Dan Feeney, DVM, from the University of Minnesota Veterinary Medical Center for their assistance with animal irradiation.

PY - 2013/9

Y1 - 2013/9

N2 - Objectives. To evaluate clinical and immunohistopathological effects of topical glycyl-histidyl-lysine-copper (GHK-Cu) on in vivo irradiated rat wounds. Design. Animal model. Setting. Academic institution. Subjects and Methods. After dorsal irradiation and a 28-day recovery period, 2 × 8 cm cranially based dorsal flaps were created in Sprague-Dawley rats. Twice daily GHK-Cu gel (test) or aquaphilic ointment (control) was applied for 10 days. Animals were euthanized, digital images of flaps were taken, and harvested tissues were immunohistochemically stained for a vascular endothelium marker, caveolin-1, and vascular endothelial growth factor (VEGF). Digital image analysis was used for outcome measures. Unpaired t-tests were used for statistical analyses; significance of P < .01 accounted for multiple comparisons. Results. By digital analysis of clinical images, 13 test and 10 control animals showed mean ischemic areas of 5.0 cm2 (SD = 0.9) for tests and 3.8 cm2 (SD = 1.1; P = .011) for controls. Whole slide digitized images allowed quantification of caveolin-1-stained blood vessels and VEGF expression in fibroblasts at the interface of healing flaps. Caveolin-1 analyses showed a mean of 209.0 vessels (SD = 111.1) and a mean vessel luminal area of 525.7 um 2 (SD = 191.0) in tests and 207.4 vessels (SD = 109.4; P = .973) and 422.8 um2 (SD = 109.7; P = .118) in controls. VEGF quantified as the percentage of pixels exceeding a colorimetric threshold, with higher fractions of positive pixels indicating more intense staining, showed a mean intensity score of 0.34 (SD = 0.19) in tests and 0.54 (SD = 0.41; P = .169) in controls. Conclusions. Irradiated dorsal rat flaps treated with topical GHK-Cu gel demonstrated no difference in flap ischemia, blood vessel number or area, or VEGF expression compared to controls.

AB - Objectives. To evaluate clinical and immunohistopathological effects of topical glycyl-histidyl-lysine-copper (GHK-Cu) on in vivo irradiated rat wounds. Design. Animal model. Setting. Academic institution. Subjects and Methods. After dorsal irradiation and a 28-day recovery period, 2 × 8 cm cranially based dorsal flaps were created in Sprague-Dawley rats. Twice daily GHK-Cu gel (test) or aquaphilic ointment (control) was applied for 10 days. Animals were euthanized, digital images of flaps were taken, and harvested tissues were immunohistochemically stained for a vascular endothelium marker, caveolin-1, and vascular endothelial growth factor (VEGF). Digital image analysis was used for outcome measures. Unpaired t-tests were used for statistical analyses; significance of P < .01 accounted for multiple comparisons. Results. By digital analysis of clinical images, 13 test and 10 control animals showed mean ischemic areas of 5.0 cm2 (SD = 0.9) for tests and 3.8 cm2 (SD = 1.1; P = .011) for controls. Whole slide digitized images allowed quantification of caveolin-1-stained blood vessels and VEGF expression in fibroblasts at the interface of healing flaps. Caveolin-1 analyses showed a mean of 209.0 vessels (SD = 111.1) and a mean vessel luminal area of 525.7 um 2 (SD = 191.0) in tests and 207.4 vessels (SD = 109.4; P = .973) and 422.8 um2 (SD = 109.7; P = .118) in controls. VEGF quantified as the percentage of pixels exceeding a colorimetric threshold, with higher fractions of positive pixels indicating more intense staining, showed a mean intensity score of 0.34 (SD = 0.19) in tests and 0.54 (SD = 0.41; P = .169) in controls. Conclusions. Irradiated dorsal rat flaps treated with topical GHK-Cu gel demonstrated no difference in flap ischemia, blood vessel number or area, or VEGF expression compared to controls.

KW - animal model

KW - copper tripeptide

KW - irradiation

KW - radiation

KW - rat

KW - wound healing

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Effects of topical copper tripeptide complex on wound healing in an irradiated rat model

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