The Function of Nitric Oxide in Wound Repair: Inhibition of Inducible Nitric Oxide-Synthase Severely Impairs Wound Reepithelialization
Recently, we demonstrated a large induction of inducible nitric oxide synthase (iNOS) during cutaneous wound repair. In this study, we established an in vivo model in mice to investigate the role of NO during the wound healing process. During excisional repair, mice were treated with L-N6-(1-iminoet...
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Published in | Journal of investigative dermatology Vol. 113; no. 6; pp. 1090 - 1098 |
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Main Authors | , , , , |
Format | Journal Article Conference Proceeding |
Language | English |
Published |
Danvers, MA
Elsevier Inc
01.12.1999
Nature Publishing |
Subjects | |
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Abstract | Recently, we demonstrated a large induction of inducible nitric oxide synthase (iNOS) during cutaneous wound repair. In this study, we established an in vivo model in mice to investigate the role of NO during the wound healing process. During excisional repair, mice were treated with L-N6-(1-iminoethyl)lysine (L-NIL), a selective inhibitor of iNOS enzymatic activity. Compared with control mice, L-NIL-treated animals were characterized by a severely impaired reepithelialization process, as the hyperproliferative epithelia at the wound edges appeared to be delayed and characterized by an atrophied morphology. Immunohistochemical labeling for detection of proliferating cells (BrdU-, Ki67-staining) revealed a strong reduction in proliferating keratinocyte cell numbers during the process of re-epithelialization after inhibition of iNOS activity during repair. Western blot analysis of total wound lysates from PBS- and L-NIL-treated mice clearly demonstrated a reduction in proliferating cell nuclear antigen, representing a marker for cell proliferation, in lysates isolated from L-NIL-treated mice. The dependency between keratinocyte proliferation and NO availability observed during wound repair in vivo is further supported by the observation that proliferation of the keratinocyte cell line (HaCaT) is stim-ulated by low concentrations of NO-donors also in vitro. In summary, our data demonstrate that the presence of a functionally active iNOS is a crucial prerequisite for normal wound reepithelialization. |
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AbstractList | Recently, we demonstrated a large induction of inducible nitric oxide synthase (iNOS) during cutaneous wound repair. In this study, we established an in vivo model in mice to investigate the role of NO during the wound healing process. During excisional repair, mice were treated with L-N6-(1-iminoethyl)lysine (L-NIL), a selective inhibitor of iNOS enzymatic activity. Compared with control mice, L-NIL-treated animals were characterized by a severely impaired reepithelialization process, as the hyperproliferative epithelia at the wound edges appeared to be delayed and characterized by an atrophied morphology. Immunohistochemical labeling for detection of proliferating cells (BrdU-, Ki67-staining) revealed a strong reduction in proliferating keratinocyte cell numbers during the process of re-epithelialization after inhibition of iNOS activity during repair. Western blot analysis of total wound lysates from PBS- and L-NIL-treated mice clearly demonstrated a reduction in proliferating cell nuclear antigen, representing a marker for cell proliferation, in lysates isolated from L-NIL-treated mice. The dependency between keratinocyte proliferation and NO availability observed during wound repair in vivo is further supported by the observation that proliferation of the keratinocyte cell line (HaCaT) is stim-ulated by low concentrations of NO-donors also in vitro. In summary, our data demonstrate that the presence of a functionally active iNOS is a crucial prerequisite for normal wound reepithelialization. Recently, we demonstrated a large induction of inducible nitric oxide synthase (iNOS) during cutaneous wound repair. In this study, we established an in vivo model in mice to investigate the role of NO during the wound healing process. During excisional repair, mice were treated with L-N6-(1-iminoethyl)lysine (L-NIL), a selective inhibitor of iNOS enzymatic activity. Compared with control mice, L-NIL-treated animals were characterized by a severely impaired reepithelialization process, as the hyperproliferative epithelia at the wound edges appeared to be delayed and characterized by an atrophied morphology. Immunohistochemical labeling for detection of proliferating cells (BrdU-, Ki67-staining) revealed a strong reduction in proliferating keratinocyte cell numbers during the process of re-epithelialization after inhibition of iNOS activity during repair. Western blot analysis of total wound lysates from PBS- and L-NIL-treated mice clearly demonstrated a reduction in proliferating cell nuclear antigen, representing a marker for cell proliferation, in lysates isolated from L-NIL-treated mice. The dependency between keratinocyte proliferation and NO availability observed during wound repair in vivo is further supported by the observation that proliferation of the keratinocyte cell line (HaCaT) is stimulated by low concentrations of NO-donors also in vitro. In summary, our data demonstrate that the presence of a functionally active iNOS is a crucial prerequisite for normal wound reepithelialization. |
Author | Kolb, Nicole Pfeilschifter, Josef Frank, Stefan Kämpfer, Heiko Stallmeyer, Birgit |
Author_xml | – sequence: 1 givenname: Birgit surname: Stallmeyer fullname: Stallmeyer, Birgit – sequence: 2 givenname: Heiko surname: Kämpfer fullname: Kämpfer, Heiko – sequence: 3 givenname: Nicole surname: Kolb fullname: Kolb, Nicole – sequence: 4 givenname: Josef surname: Pfeilschifter fullname: Pfeilschifter, Josef – sequence: 5 givenname: Stefan surname: Frank fullname: Frank, Stefan email: S.Frank@em.uni-frankfurt.de |
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Keywords | skin keratinocytes injuries epidermis Cell proliferation Skin disease Enzyme Rodentia Wound Nitric-oxide synthase Vertebrata Experimental disease Mammalia Mouse Animal Nitric oxide Keratinocyte Skin Oxidoreductases Cicatrization |
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Snippet | Recently, we demonstrated a large induction of inducible nitric oxide synthase (iNOS) during cutaneous wound repair. In this study, we established an in vivo... |
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SubjectTerms | Animals Biological and medical sciences Dermatology Endothelial Growth Factors - genetics Enzyme Inhibitors - pharmacology epidermis Epithelium - physiology Female Fibroblast Growth Factor 10 Fibroblast Growth Factor 7 Fibroblast Growth Factors Growth Substances - physiology injuries keratinocytes Keratinocytes - physiology Lymphokines - genetics Lysine - analogs & derivatives Lysine - pharmacology Medical sciences Mice Mice, Inbred BALB C Nitric Oxide - physiology Nitric Oxide Synthase - antagonists & inhibitors Nitric Oxide Synthase Type II Proliferating Cell Nuclear Antigen - analysis RNA, Messenger - analysis skin Skin involvement in other diseases. Miscellaneous. General aspects Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factors Wound Healing |
Title | The Function of Nitric Oxide in Wound Repair: Inhibition of Inducible Nitric Oxide-Synthase Severely Impairs Wound Reepithelialization |
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