[논문]Synovial Fluid Enhances Proliferation and Migration in Canine Keratocytes

Lee, Seungji; Jeong, Seong Mok; Bae, Seul-gi; Kwon, Young-sam; Yun, Sungho

doi:10.17555/jvc.2020.08.37.4.198

[국내논문] Synovial Fluid Enhances Proliferation and Migration in Canine Keratocytes 원문보기

Journal of veterinary clinics = 한국임상수의학회지, v.37 no.4, 2020년, pp.198 - 203

Lee, Seungji (Department of Veterinary Surgery, College of Veterinary Medicine, Kyungpook National University) , Jeong, Seong Mok (Department of Veterinary Surgery, College of Veterinary Medicine, Chungnam National University) , Bae, Seul-gi (Department of Veterinary Internal Medicine, College of Veterinary Medicine, Kyungpook National University) , Kwon, Young-sam (Department of Veterinary Surgery, College of Veterinary Medicine, Kyungpook National University) , Yun, Sungho (Department of Veterinary Surgery, College of Veterinary Medicine, Kyungpook National University)

Abstract ▼ AI-Helper

Synovial fluid (SF) contains various factors which may be helpful for corneal stromal healing, such as cytokines, growth factors, hyaluronic acid, and proteins. Therefore, the purpose of this study was to determine the effect of SF on proliferation and migration in canine keratocytes. In order to evaluate the degree of proliferation and migration, canine keratocytes were cultured in DMEM containing 1%, 3%, 5%, or 10% SF. Real-time PCR was performed in a control group and the group treated with 5% SF, in order to measure the expression levels of factors associated with corneal wound healing. These factors included interleukin-1α (IL-1α), hepatocyte growth factor (HGF), transforming growth factor-β (TGF-β), and α-smooth muscle actin (SMA-α). Proliferation assays demonstrated that proliferation was significantly enhanced in groups treated with greater than 3% SF, as compared with that of the control group. In addition, migration in all SF-treated groups was significantly increased as compared with migration in the control group, as measured by migration assays. mRNA expression of IL-1α and HGF was significantly increased and mRNA expression of TGF-β and SMA-α was significantly decreased in the cells treated with 5% SF. These findings suggest that SF may promote corneal wound healing.

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문제 정의

This study is the first , to our knowledge, that has assessed the response of canine keratocytes to SF treatment. Normal SF increased proliferation and migration without increases in mRNA expression of SMA-α.

제안 방법

The cells were washed with PBS and incubated for 24 hours in medium supplemented with the following: no supplement, 1% SF, 3% SF, 5% SF, or 10% SF. To quantify the area of migration, phase contrast and fluorescent images of the cell-free gap before treatment and after 12 hours of treatment were captured using a digital camera coupled to an inverted microscope (CKX41, Olympus, Japan). The area of the gap and the migratory cells were calculated using the ImageJ software (National Institutes of Health, Maryland, USA).
These results suggest that normal SF may be helpful in corneal wound healing without affecting the generation of myofibroblasts. As this study was performed in simplified keratocytes culture for only a short time, long-term in vivo studies are necessary to further evaluate the effects of SF in corneal wound healing.

대상 데이터

Preparation of canine SF

For the preparation of SF, ten healthy male Beagle dogs (five to eight years of age, 10-15 kg) were used. All used joints of dogs were corfirmed to clinically healthy with physical and radiographic examinations.

데이터처리

For statistical analysis, parametric one-way analysis of variance followed by a Fisher test was performed using SigmaPlot Version 14 (Systat Software, Inc., USA). As it is difficult to assess the assumption of normality with the small amount of data available, a non-parametric Mann-Whitney analysis test was used to test group differences in the realtime PCR analysis (SPSS Statistics version 23, IBM SPSS Inc.
, USA). As it is difficult to assess the assumption of normality with the small amount of data available, a non-parametric Mann-Whitney analysis test was used to test group differences in the realtime PCR analysis (SPSS Statistics version 23, IBM SPSS Inc., USA). A value of p ≤ 0.

성능/효과

001). A second assay after 96 hours of treatment showed increased proliferation in the groups treated with 1%, 3%, 5%, and 10% SF (Fig 1B and 1C). All SF-treated groups in the second assay were statistically significant as compared with the control group (p <0.

후속연구

This suggests that SF may help induce the wound healing process, through remodeling the wound by triggering apoptosis of injured cells and proliferation of new cells. However, because IL-1 mediates numerous inflammatory responses and pathological changes in various organs, further in vivo work is needed to further evaluate the effects of IL-1 induction by SF.
This result suggests that SF may contribute to corneal wound healing by enhancing proliferation of epithelial cells through HGF. However, as epithelial wound healing is mediated by numerous cytokines, additional work is needed to further identify the effects of SF in epithelial cells of the cornea.
However, the TGF-β that has an effect on keratocytes is predominantly secreted from epithelial cells. Further studies are needed to observe the effects of SF on corneal wound healing in vivo.
This result suggests that SF accelerates migration of keratocytes but does not increase numbers of myofibroblasts. However, as corneal haze is generally considered a long-term complication of a deep corneal wound, further long-term studies are needed to better assess this phenotype.

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