In this study, we aimed at fabricating decellularized bovine myocardial extracellular matrix-based films (dMEbF) for cardiac tissue engineering (CTE). The decellularization process was carried out utilizing four consecutive stages including hypotonic treatment, detergent treatment, enzymatic digestion and decontamination, respectively. In order to fabricate the dMEbF, dBM were digested with pepsin and gelation process was conducted. dMEbF were then crosslinked with N-hydroxysuccinimide/1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (NHS/EDC) to increase their durability. Nuclear contents of native BM and decellularized BM (dBM) tissues were determined with DNA content analysis and agarose-gel electrophoresis. Cell viability on dMEbF for 3rd, 7th, and 14th days was assessed by MTT assay. Cell attachment on dMEbF was also studied by scanning electron microscopy. Trans-differentiation capacity of human adipose-derived mesenchymal stem cells (hAMSCs) into cardiomyocyte-like cells on dMEbF were also evaluated by histochemical and immunohistochemical analyses. DNA contents for native and dBM were, respectively, found as 886.11 ± 164.85 and 47.66 ± 0.09 ng/mg dry weight, indicating a successful decellularization process. The results of glycosaminoglycan and hydroxyproline assay, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), performed in order to characterize the extracellular matrix (ECM) composition of native and dBM tissue, showed that the BM matrix was not damaged during the proposed method. Lastly, regarding the histological study, dMEbF not only mimics native ECM, but also induces the stem cells into cardiomyocyte-like cells phenotype which brings it the potential of use in CTE.
Ö Z B u çalışmada, Melia azedarach L. yeşil meyve ve yapraklarının sulu özleri iki farklı özütleme yöntemi kullanılarak elde edildi. Yeşil meyve ve yaprakların özütleme verimleri sırasıyla, demleme yöntemi için %24.11 ve %37.98; çalkalama yöntemi için ise %17.76 ve %27.00 olarak bulundu. Demleme yöntemi ile ilgili toplam fenolik içerik, yeşil meyve özü için 173.67±10.84 mg galik asit eşdeğeri (GAE)/g kuru ağırlık ve yaprak özü için 312.33±9.81 mg GAE/g kuru ağırlık cinsinden tespit edildi. Diğer yandan demleme yönteminde, yeşil meyve özü için 172.51±13.23 mg Trolox/L ve yaprak özü için ise 569.16±10.41 mg Trolox/L cinsinden antioksidan aktivite belirlendi. Özütlerin kimyasal bileşimini tanımlamak için gaz kromatografisi-kütle spektrometresi (GC-MS) analizi yapıldı. Özütlerin insan adipoz kaynaklı mezenkimal kök hücreleri (iAMKH'leri) üzerindeki sitotoksisite seviyeleri ticari olarak mevcut olan XTT testi ile değerlendirildi. Sonuç olarak, yeşil meyve özütünün iAMKH'leri üzerine yaprak özütünden daha fazla sitotoksik aktiviteye sahip olduğu bulunmuştur Anahtar KelimelerMelia azedarach L., toplam fenolik bileşikler, sitotoksisite, insan adipoz kaynaklı mezenkimal kök hücreler. A B S T R A C TI n this study, aqueous extracts of Melia azedarach L. green fruit and leaves were obtained using two different extraction methods. The extraction yields of the green fruit and leaves were found as 24.11% and 37.98% for the infusion method; 17.76% and 27.00% for the rotating method, respectively. The total phenolic content, related to the infusion method, was ascertained for green fruit extract 173.67±10.84 mg Gallic Acid Equivalent (GAE)/g dry weight and leaf extract 312.33±9.81 mg GAE/g dry weight. In other respects, antioxidant activity related to the infusion method was determined for green fruit extract 172.51±13.23 mg Trolox/L and leaf extract 569.16±10.41 mg Trolox/L. Gas chromatography-mass spectrometry (GC-MS) analysis was performed to identify the chemical composition of the extracts. The cytotoxicity levels of the extracts were assessed on human adipose-derived mesenchymal stem cells (hAMSCs) using commercially available XTT assay. Consequently, it has been found that the green fruit extract has more cytotoxic activity than the leaf extract on hAMSCs.
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