In Vitro Vitamin K3 Effect on Conjunctival Fibroblast Migration and Proli… — Volkswagen Polo Lim

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In Vitro Vitamin K 3 Effect on Fibroblast Migration and Proliferation

1 of Ophthalmology, Lozano Blesa Hospital, C/San Juan 15, 50009 Zaragoza, Spain

2 Institute of Health Sciences Aragon), 50009 Zaragoza,

3 Department of Ophthalmology, Hospital Orcoyen, Navarra, 31200 Spain

Received 29 August Accepted 14 October 2013; 8 January 2014

Academic J. Aquavella and K. Unlu

Copyright 2014 I. Pinilla et al. This is an access article distributed the Creative Commons Attribution which permits unrestricted distribution, and reproduction in any medium, the original work is properly


Purpose . To evaluate the effect of vitamin K 3 on wound mechanisms. Methods . Conjunctival were incubated for 24 hours. An wound was made and the cells incubated with fresh plus doses of vitamin K 3 to be Wound repair was monitored at 0, 18, 24, and 48 Proliferation was measured in actively cells by [ 3 H]thymidine uptake. Six groups were tested: 1/no drugs added, 2/ethanol 0.1#x25;, group K 3 1#x2009;mg/L, group 4/vitamin K 3 group 5/vitamin K 3 4#x2009;mg/L, and 6/vitamin K 3 6#x2009;mg/L. Each was carried out in triplicate and 4 times. . There were no differences groups at the initial time. In wound repair was slower in 4, 5, and 6. There were no differences control and ethanol groups and control and vitamin K 3 1 mg/L Fibroblast mitogenic activity was decreased in all vitamin K groups;

differences were found vitamin K 3 1#x2009;mg/mL and higher too. In groups 5 and 6, cellular was presented. Conclusions . Vitamin K 3 is to inhibit fibroblast proliferation. K 3 2#x2009;mg/L or higher doses wound healing repair, cellular toxicity at 4 and 6#x2009;mg/L.

1. Introduction

Antimetabolites and other inhibitor drugs have shown to enhance the success of filtering surgery although, on the dose, they can lead to complications and may result in the failure of the

Corticosteroids [1 #x2013;6 ] antiproliferative (5-fluoro-uracil and other fluoropyrimidines, doxorubicin, mycophenolate mofetil#x2026;, or in combination or with different systems) [3 #x2013;12 ], systemic, intraocular steroidal, and nonsteroidal agents [5. 13 #x2013;16 ], colchicine [8 ], [8 ], tissue plasminogen activator [17 ], [12. 18 #x2013;20 ], interferon-gamma 22 ], calcium channel blockers [23 ], and lysyl hydroxylase inhibitors 24 #x2013;26 ], retinoic acid 28 ], alpha-tocopherol [29 #x2013;31 ], disintegrins [32 ], #x3b1; [33 ]#x2026; are some of the drugs that have used in the treatment of conditions as proliferative vitreoretinopathy, bleb after trabeculectomy, and other with cell proliferation conjunctival or extraocular cicatrization).

Vitamin K 3 (menadione, 2-methyl-1,4-naphthoquinone) has used as antihemorrhagic agent. Its to inhibit proliferation of tumor has already been reported; its has been demonstrated in human stem cell and it is used in trial for advanced malignancies in different pathways and has also related to other oxidative processes at the eye level as cataract [34 #x2013;40 ]. Liu et al. reported that drug could inhibit of rabbit conjunctive cells [41 ].

The aim of this study was to evaluate and to the antiproliferative properties of vitamin K 3 in human fibroblasts.

2. Methods


All supplies for cell were purchased from (Roskilde, DK). Dulbecco#x2019;s Eagles Medium (DMEM), buffer saline (PBS), calf serum (FCS), and were purchased from (Madison, WI). [methyl- 3 was purchased from Amershm (Madrid, Spain). 2-Methyl (Menadione) (98#x25;) was obtained Sigma (St. Louis, The drug was initially dissolved in ethanol. This alcohol was then diluted into BSS to a final ethanol concentration of

2.2. Cell Cultures

fibroblasts were obtained explants of a healthy … who underwent ophthalmic surgery for detachment. All subjects gave consent to participate in the study, was conducted in accordance with the of the Declaration of Helsinki, and the experimental was approved by the local Ethics of the Aragon Health Science Cells were cultured in plastic flasks in DMEM with antibiotics and antifungals penicillin, 100#x2009; #x3bc; streptomycin, and 0.25#x2009; #x3bc; amphotericin B), and 20#x25; fetal serum (FCS) in a humidified at 37 degrees Celsius and 5#x25; CO 2 . The medium was changed every 3 day and the were performed with obtained between the 5th and 8th passages.

2.3. Wounding Assays

assays were performed the method described by Sato and [42 ]. An artificial wound was made by cell denudation with a tip, as described in previous [26 ]. The wound repair process was by two independent observers measuring the area (mm 2 ) in a blind fashion, at times: 0, 18, 24, and 48 hours (Figure 1 ). The area was quantified in the elliptic or shape wounds with size. Then, the major and the axes were measured in a microscopy equipped with a visor. The area was calculated the mathematical formula: area =

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