Perikardial Sıvıdan İzole Edilen Hücrelere Fibroblast Büyüme Faktörü (FGF-2) Etkisinin Analizi

Yıl 2020, , 1156 - 1164, 31.12.2020

Dilek Sönmezer , Güler Toprak , Ayhan Düzler , Fatma Latifoğlu

https://doi.org/10.35414/akufemubid.654177

Öz

Perikardiyal sıvı (PF) heterojen bir hücre topluluğuna sahiptir. Bu hücrelerden biri de mezenkimal kök hücre özelliği taşıyarak başka hücrelere farklılaşma potansiyeline sahip olan mesotelyal hücrelerdir. Bu hücrelerin sayısı doku hasarıyla birlikte artarak büyüme faktörleri (VEGF, FGF-2, PDGF, TGF-b) ve ekstrasellüler matriks (ECM) moleküllerini sentezleyerek doku/organda meydana gelen hasarın onarımında önemli bir yer tutmaktadır. Bu çalışmada mesotelyal hücrelerin hücre kültür ortamına FGF-2 (basic fibroblast growth factor)’ nin eklenmesiyle mesotelyal hücrelerin vasküler doku oluşumuna etkisi incelenmiştir. Bu amaçla büyüme faktörü eklenmiş ve büyüme faktörü eklenmemiş olan kontrol grubu hücrelerinin birinci ve beşinci günlerdeki ışık mikroskobu görüntüleri kullanılarak analiz edilmiştir. Bu görüntüler ve görüntü histogramları kullanılarak Hjorth parametreleri olan mobilite, aktivite ve komplekslik özellikleri, çarpıklık, basıklık ve entropi değerleri kullanılarak istatistiksel olarak analiz edilmiştir. Birinci ve beşinci günlerde FGF-2 eklenen ve eklenmeyen kültürdeki hücrelerin görüntülerinden hesaplanan özniteliklerin gruplar arasındaki farklılığı değerlendirmek için istatistiksel analizi yapılmıştır. Elde edilen sonuçlara göre birinci günde iki grup arasında ve beşinci günde iki grup arasında bağımsız örneklem t testi ve Mann-Whitney U testi neticesinde anlamlı farklılıklar olduğu (p<0.05) görülmüştür. Doku hasarının giderilmesi için gerekli olan ECM bileşenleri ve büyüme faktörlerini sentezleyebilen mezotelyal hücrelerin literatürde yaygın olarak kullanılan mezenkimal kök hücreye alternatif olarak kullanılabileceği bu ön çalışma ile ortaya konulmuştur.

Anahtar Kelimeler

Perikardiyal sıvı, Mezotelyal hücre, bGFG (FGF2), Hjorth parametreleri, Doku mühendisliği

Destekleyen Kurum

Erciyes Üniversitesi, Bilimsel Araştırmalar Projeleri Birimi (Bap)

Proje Numarası

FOA-2016-6692

Teşekkür

Bu çalışma Erciyes Üniversitesi, Bilimsel Araştırmalar Projeleri Birimi (Bap) ÖNAP, FOA-2016-6692 kodlu proje ile desteklenmiştir. Deney çalışmaları Erciyes Üniversitesi, Genom ve Kök Hücre Merkezi (GENKÖK) gerçekleştirilmiş olup katkılarından dolayı teşekkür ederiz.

Analysis of Fibroblast Growth Factor (FGF-2) Effect on Cells Isolated from Pericardial Fluid

Öz

Pericardial fluid (PF) has a heterogeneous cell population. One of these cells is mesothelial cells that have the potential to differentiate into other cells, such as mesenchymal stem cells. The number of these cells increases with tissue damage and plays an important role in the repair of tissue/organ damage by synthesizing growth factors (VEGF, FGF-2, PDGF, TGF-b) and extracellular matrix (ECM) molecules. In this study, the effect of mesothelial cells on vascular tissue formation was investigated by adding FGF-2 (basic fibroblast Görowth factor) to cell culture medium. For this purpose, control group cells and groups of added growth factor were analyzed using light microscopy images at the first and fifth days. Using these images and image histograms, Hjorth parameters which are mobility, activity, complexity and, kurtosis, skewness and entropy values were analyzed statistically. On the first and fifth days, statistical analysis was performed to evaluate the differences between the groups calculated from the images of cells in culture with FGF-2 and without FGF-2 addition. According to the results of independent samples t test and Mann-Whitney U test showed significant differences between the two groups on the first day and between the two groups on the fifth day (p <0.05).
It was demonstrated with this preliminary study that mesothelial cells, which can synthesize ECM components and growth factors required for tissue damage, can be used as an alternative to mesenchymal stem cells the commonly used in tissue engineering applications.

Anahtar Kelimeler

Pericardial Fluid, Mesothelial Cell, Basic Fibroblast Growth Factor (bGFG /FGF2), Hjorth Parameter, Tissue Engineering

Proje Numarası

FOA-2016-6692

Kaynakça

• Abdul Alim, O., Moselhy, M., Mroueh, F.,2012. EMG Signal Processing and Diagnostic of Muscle Diseases, 2nd International Conference on Advances in Computational Tools for Engineering Applications (ACTEA), 2012, 1-6, Beirut, Lebanon, 12-15 Dec.
• Cano, M.E., Class, Q.A. Polich, J., 2009. Affective valance, stimulus attributes, and P300: Color vs. black/White and normal vs. scrambled images, International Journal of Psychophysiology, Vol. 71, , 17-24. Cecchin, T., Ranta, R., Koessler, L., Caspary, O., Vespignani, H., Maillard, L., 2010. Seizure lateralization in scalp EEG using hjorth parameters, Clinical neurophysiology, 121, 290–300.
• Dauleh, S., Santeramo, I., Fielding, C., Ward, K., Herrmann, A., Murray, P., Wilm, B. 2016. Characterisation of cultured mesothelial cells derived from the murine adult omentum. PloS one, 11(7), e0158997. Esch, F., Baird, A., Ling, N., et al, 1985. Primary structure of bovine pituitary basic fibroblast growth factor (FGF) and comparison with the amino-terminal sequence of bovine brain acidic FGF. Proceedings of the National Academy of Sciences, 82:6507-6511
• Fujita, M., Ikemoto, M., Kishishita, M. et al. 1996. Elevated basic fibroblast growth factor in pericardial fluid of patients with unstable angina. Circulation 94:610–3.
• Fujita, M., Ikemoto, M., Tanaka, T. et al. 1998. Marked elevation of vascular endothelial growth factor and basic fibroblast growth factor in pericardial fluid of patients with angina pectoris. Angiogenesis;2:105–8.
• Gorin, C., Rochefort, G.Y., Bascetin, R., Ying, H., Lesieur, J., Sadoine, J., Beckouche, N., Berndt, S., Novais, A., Lesage, M., Hosten, B., Vercellino, L., Merlet, P., Le-Denmat, D., Marchiol, C., Letourneur, D., Nicoletti, A., Vital, S.O., Poliard, A., Salmon, B., Muller, L., Chaussain, C., Germain, S., 2016. Priming Dental Pulp Stem Cells With Fibroblast Growth Factor-2 Increases Angiogenesis of Implanted Tissue-Engineered Constructs Through Hepatocyte Growth Factor and Vascular Endothelial Growth Factor Secretion. Stem Cells Translational Medicine, Mar;5(3):392-404.
• Laham, R. J., Simons, M., Tofukuji M., Hung, D., Sellke, F.W., 1998. Modulation of myocardial perfusion and vascular reactivity by pericardial basic fibroblast growth factor: Insight into ischemia-induced reduction in endothelium-dependent vasodilatation. Journal of Thoracic and Cardiovascular Surgery, 116:1022–8 Landau, C., Jacobs, A.K., Haudenschild, C.C. 1995. Intrapericardial basic fibroblast growth factor induces myocardial angiogenesis in a rabbit model of chronic ischemia. American Heart Journal, 129:924–31.
• Latifoğlu, F., Sönmezer, D.,Toprak, G., Düzler A., İşoğlu, İ.A., Kaan, D., 2018. Cell isolating from bovine pericardial fluid and culturing for next tissue engineering applications. International Conference on Technology, Engineering and Science (Icontes 2018), Antalya, Türkiye, 26-29 October, cilt.4, ss.224-229
• Mahaphonchaikul, K., Sueaseenak, D., Pintavirooj, C., Sangworasil, M., 2010. EMG signal feature extraction based on wavelet transform, International Conferance on Electrical Engineering/Electronics Computer Telecommunications and Information Technology (ECTI-CON), Vol. 1, 356–360, Chiang Mai, Thailand, 19-21 May.
• Ornitz, D.M., Itoh, N., 2015. The Fibroblast Growth Factor signaling pathway. Wiley Interdiscip. Rev. Dev. Biol., 4, 215–266.
• Rozenkrants, B. and Polich, J., 2008. Affective ERP processing in a visual oddball task: Arousal, valance, and, Clinical Neurophysiology, Vol. 119, 2260-2265.
• Rozenkrants, B., Olofsson, J.K. Polich, J., 2008. Affective visual event-related potentials: Arousal, valance, and repetition effects for normal and distorted pictures, International Journal of Psychophysiology, Vol. 67, 2008, 114-123
• Sanei, S., Hosseini-Yazdi, A., 2011. Extraction of ECG from single channel EMG signal using constrained singular spectrum analysis, 17th International Conference on Digital Signal Processing (DSP), 1-4, Corfu, Greece, 6-8 July
• Schelch, K., Wagner, C., Hager, S., Pirker, C., Siess, K., Lang, E., Marian, B., 2018. FGF2 and EGF induce epithelial–mesenchymal transition in malignant pleural mesothelioma cells via a MAPKinase/MMP1 signal. Carcinogenesis, 39(4), 534-545.
• Sönmezer, D., Latifoğlu, F., İşoğlu, İ.A., Düzler, A., Toprak, G., Kaan, D., 2016. Hücre Bileşeni ve Biyomalzeme ile Vasküler Doku Üretimi. Tıp Teknolojileri Ulusal Kongresi (TIPTEKNO'16), Antalya, Türkıye, ss.205-208, 27-29 Ekim
• Uchida, Y., Yanagisawa-Miwa, A., Nakamura, F., Yamada, K., Tomaru, T., Kimura, K., Morita, T.. 1995. Angiogenic therapy of acute myocardial infarction by intrapericardial injection of basic fibroblast growth factor and heparin sulfate: an experimental study. American Heart Journal, 130:1182-1188. Vasfi, T.N., 2014. SPSS Uygulamalı İstatistik Teknikleri, 3. Baskı, 296 Syf.
• Yu, P.J., Ferrari, G., Galloway, A.C., Mignatti, P., Pintucci, G., 2007. Basic fibroblast growth factor (FGF-2): the high molecular weight forms come of age. Journal of Cellular Biochemistry, 100: 1100–1108.