scholarly journals The opioid peptide biphalin modulates human corneal epithelial wound healing in vitro

2019 ◽  
Author(s):  
Ozgun Melike Gedar Totuk ◽  
Erdost YILDIZ ◽  
Adriano MOLLICA ◽  
Kerem KABADAYI ◽  
Afsun SAHIN
Keyword(s):  
Wound Healing ◽  
Corneal Epithelium ◽  
Wound Closure ◽  
Opioid Peptide ◽  
Corneal Epithelial Cell ◽  
Scratch Assay ◽  
Corneal Epithelial ◽  
Significant Difference ◽  
Epithelial Wound Healing

Abstract Background Analgesic drugs including nonselective opioids and non-steroidal anti-inflammatory drugs should be used with great precautions to relieve the pain after physical damage of the corneal epithelium, because of their unfavorable effects on wound healing process. Biphalin, a synthetic opioid peptide, which has been demonstrated to possesses a strong analgesic effect on rodents. The purpose of this study is to investigate the effects of biphalin on human corneal epithelium wound healing. Methods Immortalized human corneal epithelial cell (HCEC) culture was used to test the effects of biphalin on wound healing. The toxicity of biphalin in various concentrations was measured with MTT assay. The effect of 1 µM and 10 µM biphalin were tested on wound closure at in vitro scratch assay of HCECs, and for cell migration and proliferation separately. Naloxone, a non-selective competitive antagonist of opioid receptor, was also used to inhibit the effects of biphalin in all experiments. Results Biphalin did not cause any toxic effect on HCECs in concentrations lower than 100 µM at various incubation time points. Biphalin increased wound closure process significantly at 1 µM concentration at in vitro scratch assay of HCECs (p < 0.05); also increased migration of HCECs significantly (p < 0.01). There was no significant difference between biphalin and control groups of HCECs at Ki67 proliferation assay. Conclusion Biphalin, a synthetic opioid peptide, has a potential role as a novel topical analgesic agent that promotes corneal epithelial wound healing.

scholarly journals The opioid peptide biphalin modulates human corneal epithelial wound healing in vitro

2020 ◽  
Author(s):  
Ozgun Melike Gedar Totuk ◽  
Erdost YILDIZ ◽  
Adriano MOLLICA ◽  
Kerem KABADAYI ◽  
Afsun SAHIN
Keyword(s):  
Wound Healing ◽  
Corneal Epithelium ◽  
Wound Closure ◽  
Opioid Peptide ◽  
Corneal Epithelial Cell ◽  
Scratch Assay ◽  
Corneal Epithelial ◽  
Significant Difference ◽  
Epithelial Wound Healing

Abstract Background: Analgesic drugs, including nonselective opioids and non-steroidal anti-inflammatory drugs, should be used with great precautions to relieve pain after physical damage of the corneal epithelium because of their unfavorable effects on the wound-healing process. Biphalin is a synthetic opioid peptide that has been demonstrated to possesses a strong analgesic effect on rodents. The purpose of this study is to investigate the effects of biphalin on human corneal epithelium wound healing.Methods: An immortalized human corneal epithelial cell (HCEC) culture was used to test the effects of biphalin on wound healing. The toxicity of biphalin in various concentrations was measured with the MTT assay. The effect of 1 µM and 10 µM biphalin were tested on wound closure in an in vitro scratch assay of HCECs and for cell migration and proliferation separately. Naloxone, a non-selective competitive antagonist of opioid receptors, was also used to inhibit the effects of biphalin in all experiments.Results: Biphalin did not cause any toxic effect on HCECs in concentrations lower than 100 µM at various incubation time points. Biphalin increased the wound closure process significantly at 1 µM concentration in an in vitro scratch assay of HCECs (p < 0.05). It also increased the migration of HCECs significantly (p < 0.01). There was no significant difference between biphalin and control groups of HCECs in the Ki67 proliferation assay.Conclusion: Biphalin, a synthetic opioid peptide, has a potential role as a novel topical analgesic agent that promotes corneal epithelial wound healing. This role should be evaluated in further in vivo and clinical studies.

scholarly journals Effects of Biphalin on Corneal Epithelial Wound Healing

Proceedings ◽  
2018 ◽  
Vol 2 (25) ◽  
pp. 1552 ◽  
Author(s):  
Erdost Yıldız ◽  
Özgün Melike Gedar Totuk ◽  
Adriano Mollica ◽  
Kerem Kabadayı ◽  
Afsun Şahin
Keyword(s):  
Wound Healing ◽  
Mtt Assay ◽  
Corneal Epithelium ◽  
Healing Process ◽  
Wound Healing Process ◽  
Corneal Epithelial ◽  
Positive Effects ◽  
Study Results ◽  
Significant Difference ◽  
Epithelial Wound Healing

After physical or surgical damage of corneal epithelium, most of analgesic drugs, like non-selective opioid agonists and non-steroid anti-inflammatory drugs, cannot be used because of their negative effects on wound healing process. Biphalin is selective µ and Δ opioid receptor agonist which has proven analgesic effects on rodents. Our purpose of study is finding effects of biphalin on wound healing of corneal epithelium. We used primary culture of human corneal epithelial cells (HCECs) for examining effects of biphalin on wound healing. Firstly, we measured toxicity of Biphalin in various concentrations with MTT assay and we showed biphalin has no toxic effects on HCECs in lower concentrations than 100 µM in various incubation times. After MTT assay, we administered 1 µM and 10 µM biphalin at in vitro scratch assay of HCECs, biphalin increased wound closure process significantly at 1 µM concentration (p < 0.05). Then we tested effects of biphalin on cell migration and proliferation separately. Bifalin increased migration of HCECs significantly (p < 0.01) at transwell migration assay. But we did not observe any significant difference between groups in Ki67 proliferation assay. In all these experiments, we also used naloxone to inhibiting effects of biphalin. In biphalin plus naloxone groups, effects of biphalin decrease partially. Our study results suggest, biphalin has positive effects on epithelial wound healing via opioid receptors. This effect because of increased migration of HCECs under influence of biphalin. With these findings, we propose biphalin as a new analgesic agent for post-surgical and post-traumatic care of corneal epithelial wounds.

Effect of Ofloxacin on Corneal Epithelial Wound Healing Evaluated by In Vitro and In Vivo Methods

1993 ◽  
Vol 6 (2) ◽  
pp. 96-103 ◽  
Author(s):  
Steven S. Matsumoto ◽  
Michael E. Stern ◽  
Roger M. Oda ◽  
Corine R. Ghosn ◽  
Josephine W. Cheng ◽  
...  
Keyword(s):  
Wound Healing ◽  
Corneal Epithelial ◽  
Epithelial Wound Healing

scholarly journals Heparin-Modified Amniotic Membrane Combined With Growth Factors for Promoting Corneal Wound Healing After Alkali Burn

2020 ◽  
Vol 8 ◽  
Author(s):  
Xuan Zhao ◽  
Xin Zuo ◽  
Jing Zhong ◽  
Bowen Wang ◽  
Saiqun Li ◽  
...  
Keyword(s):  
Wound Healing ◽  
Growth Factors ◽  
Sustained Release ◽  
Corneal Epithelial Cell ◽  
Control Group ◽  
Therapeutic Effects ◽  
Cell Growth And Migration ◽  
Corneal Epithelial

Ocular chemical burns are potentially blinding ocular injuries and require urgent management. Amniotic membrane (AM) transplantation is an effective surgical treatment, one of the reasons is because AM is a rich source of growth factors that can promote epithelialization and wound healing. However, growth factors will be gradually lost and insufficient after preparation process and long-time storage, leading to unsatisfactory therapeutic effects. Herein, we present a modified AM (AM-HEP) for the supplement and sustained release of growth factor by surface grafting heparin for treatment of ocular chemical burns. Heparin grafting rate and stability, microstructure, physical property, and sustained release of epithelial growth factor (EGF) of AM-HEP were characterized. Biocompatibility and ability to promote corneal epithelial cell growth and migration were evaluated and compared with a biological amnion, which is available on the market in vitro. The therapeutic effects of AM-HEP combined with EGF (AM-HEP@EGF) in vivo had been evaluated in a model of mouse corneal alkali burn. The results indicated that heparin was introduced into AM and maintain stability over 3 weeks at 37°C. The modification process of AM-HEP did not affect microstructure and physical property after comparing with non-modified AM. EGF could be combined quickly and effectively with AM-HEP; the sustained release could last for more than 14 days. AM-HEP@EGF could significantly promote corneal epithelial cell growth and migration, compared with non-modified AM and control group. Faster corneal epithelialization was observed with the transplantation of AM-HEP@EGF in vivo, compared with the untreated control group. The corneas in the AM-HEP@EGF group have less inflammation and were more transparent than those in the control group. The results from in vitro and in vivo experiments demonstrated that AM-HEP@EGF could significantly enhance the therapeutic effects. Taken together, AM-HEP@EGF is exhibited to be a potent clinical application in corneal alkali burns through accelerating corneal epithelial wound healing.

Promotion of Corneal Epithelial Wound Healing In Vitro and In Vivo by Annexin A5

2006 ◽  
Vol 47 (5) ◽  
pp. 1862 ◽  
Author(s):  
Masanao Watanabe ◽  
Shoichi Kondo ◽  
Ken Mizuno ◽  
Wataru Yano ◽  
Hiroshi Nakao ◽  
...  
Keyword(s):  
Wound Healing ◽  
Annexin A5 ◽  
Corneal Epithelial ◽  
Epithelial Wound Healing

Soluble and insoluble fibronectin increases alveolar epithelial wound healing in vitro

1996 ◽  
Vol 271 (5) ◽  
pp. L844-L853 ◽  
Author(s):  
C. Garat ◽  
F. Kheradmand ◽  
K. H. Albertine ◽  
H. G. Folkesson ◽  
M. A. Matthay
Keyword(s):  
Wound Healing ◽  
Wound Closure ◽  
Type Iv Collagen ◽  
Type I ◽  
Serum Free Medium ◽  
Free Medium ◽  
Alveolar Epithelial ◽  
Type Iv ◽  
Epithelial Wound Healing

Adhesive interactions between cells and extracellular matrix proteins are important in cell attachment, migration, and proliferation. The present work defines the role of fibronectin (soluble and insoluble) compared with type I and type IV collagen on in vitro alveolar epithelial wound healing. Repeated video microscopy experiments demonstrated that the half-time of wound closure was decreased in the presence of soluble fibronectin (6.6 +/- 2.1 vs. 17.4 +/- 0.8 h in serum-free medium, P < 0.05). Video microscopy, electron microscopy, and vinculin distribution demonstrated the contribution of two main events during the repair process: the migration of epithelial cell sheets and the spreading of the cells. During the wound healing, the internuclear distance between two adjacent cells at the migrating edge of the wound was significantly increased 10 h after wounding in the presence of soluble fibronectin (67 +/- 3.0 vs. 45 +/- 1.5 microns in serum-free medium, P < 0.05), indicating that cell spreading is involved as part of the mechanism for wound closure. Compared with type I and type IV collagen, insoluble fibronectin was the most potent stimulus for alveolar type II cell motility and wound healing in the absence of other serum factors. These results demonstrate that alveolar epithelial wound healing can be modulated in vitro by the composition of the extracellular matrix, an effect that may be mediated by changes in cell shape.

scholarly journals Rho kinases regulate corneal epithelial wound healing

2008 ◽  
Vol 295 (2) ◽  
pp. C378-C387 ◽  
Author(s):  
Jia Yin ◽  
Fu-Shin X. Yu
Keyword(s):  
Cell Proliferation ◽  
Wound Healing ◽  
Cell Migration ◽  
Rho Gtpase ◽  
Adhesion Formation ◽  
Small Gtpase ◽  
Corneal Epithelial Cell ◽  
Corneal Epithelial ◽  
Epithelial Wound Healing ◽  
Rho Kinases

We have previously shown that Rho small GTPase is required for modulating both cell migration and proliferation through cytoskeleton reorganization and focal adhesion formation in response to wounding. In the present study, we investigated the role of Rho kinases (ROCKs), major effectors of Rho GTPase, in mediating corneal epithelial wound healing. Both ROCK 1 and 2 were expressed and activated in THCE cells, an SV40-immortalized human corneal epithelial cell (HCEC) line, in response to wounding, lysophosphatidic acid, and heparin-binding EGF-like growth factor (HB-EGF) stimulations. The ROCK inhibitor Y-27632 efficiently antagonized ROCK activities without affecting Rho activation in wounded HCECs. Y-27632 promoted basal and HB-EGF-enhanced scratch wound healing and enhanced cell migration and adhesion to matrices, while retarded HB-EGF induced cell proliferation. E-cadherin- and β-catenin-mediated cell-cell junction and actin cytoskeleton organization were disrupted by Y-27632. Y-27632 impaired the formation and maintenance of tight junction barriers indicated by decreased trans-epithelial resistance and disrupted occludin staining. We conclude that ROCK activities enhance cell proliferation, promote epithelial differentiation, but negatively modulate cell migration and cell adhesion and therefore play a role in regulating corneal epithelial wound healing.

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