Publication: İndigo naturalis ekstresinin insan periodontal hücreleri üzerine etkisinin incelenmesi : in vitro çalışma
Abstract
Amaç: Çalışmamızda İndigo naturalis (İN) ekstresinin periodontal rejenerasyondaki olası etkinliğini değerlendirmek amacıyla insan dişeti fibroblastı (İDF), insan periodontal ligament fibroblastı (İPLF) ve insan osteoblast (İO) hücrelerinin canlılığı ve bu hücrelerin tip 1 kolajen, kemik morfogenik protein-2 (KMP-2) ve aksin-2 salınımı üzerindeki etkisinin incelenmesi hedeflenmiştir. Gereç ve Yöntem: Çalışmamız dahilinde İN ekstresi yüksek performanslı sıvı kromatografisi (YPSK) ile analiz edildi. 5, 10 ve 15 μg/ ml olmak üzere farklı konsantrasyonları hazırlanarak İDF, İPLF ve İO hücre dizilerine 24 saat süreyle uygulama sonrası MTT testi ile canlılık değerlendirmesi yapıldı. Daha sonra İPLF ve İO hücrelerine 5 μg/ ml konsantrasyon dozu uygulanarak 24 saatte meydana gelen tip 1 kolajen, KMP-2 ve aksin-2 proteinlerinin ekspresyon seviyelerini değerlendirmek amacıyla ELISA analizi yapıldı. Sonuçların istatistiksel değerlendirmesi için SPSS paket kullanıldı. Bulgular: İN uygulanan konsantrasyonlarda İDF, İPLF ve İO hücrelerinin canlılığı üzerine anlamlı etki göstermedi (p>0.05). İPLF hücrelerine 5 μg/ ml konsantrasyonda İN uygulaması, tip 1 kolajen ve KMP-2 salınımına anlamlı etki etmezken (p>0.05) aksin-2 salınımını anlamlı derecede artırdı (p=0,01). İO hücrelerine 5 μg/ ml konsantrasyonda İN uygulamasının, tip 1 kolajen, KMP-2 ve aksin-2 salınımını anlamlı derecede artırdığı tespit edildi (p=0,01, p=0,01, p=0,01). Sonuç: Bu araştırmanın sınırları dahilinde elde edilen veriler, farklı konsantrasyonlarda uygulanan İN’nin İDF, İPLF ve İO hücrelerinin canlılığına anlamlı etki göstermeyecek şekilde benzer etki gösterdiğini ortaya koymuştur. Buna ek olarak İN’nin İO hücreleri üzerinde protein ekspresyonu açısından olumlu etkileri olabileceği ve bu etkinin Wnt sinyal yolununun aktive edilmesiyle gerçekleşebileceği düşünülebilir. İN’nin farklı konsantrasyonlarda uygulandığı ve farklı takip süreleri ile değerlendirildiği çalışmalara gereksinim vardır.
Objective: The aim of our study is to evaluate the possible efficacy of Indigo naturalis (IN) extract in periodontal regeneration by examining the viability of human gingival fibroblast (HGF), human periodontal ligament fibroblast (HPLF), and human osteoblast (HO) cells, as well as the effect of this extract on the secretion of type 1 collagen, bone morphogenetic protein-2 (BMP-2), and axin-2 in these cells. Materials and Methods: In our study, IN extract was analyzed using high-performance liquid chromatography (HPLC). Different concentrations of 5, 10, and 15 μg/ ml were prepared and applied to HGF, HPLF, and HO cell lines for 24 hours, followed by viability assessment using the MTT assay. Subsequently, a concentration of 5 μg/ ml was applied to HPLF and HO cells to evaluate the expression levels of type 1 collagen, BMP-2, and axin-2 proteins at 24 hours using the ELISA analysis. SPSS software package was used for statistical evaluation of the results. Results: IN applied at the tested concentrations did not show a significant effect on the viability of HGF, HPLF, and HO cells (p>0.05). The application of 5 μg/ ml IN to HPLF cells did not significantly affect the secretion of type 1 collagen and BMP-2 (p>0.05), but significantly increased the secretion of actin-2 (p=0.01). The application of 5 μg/ ml IN to HO cells significantly increased the secretion of type 1 collagen, BMP-2, and axin-2 (p=0.01, p=0.01, p=0.01). Conclusion: Within the limits of this research, the data obtained revealed that IN applied at different concentrations exhibited a similar effect that did not significantly impact the viability of HGF, HPLF, and HO cells. Additionally, IN may have positive effects on protein expression in HO cells, potentially through the activation of the Wnt signaling pathway. Further studies are needed where IN is applied at different concentrations and evaluated over varying follow-up periods.
Objective: The aim of our study is to evaluate the possible efficacy of Indigo naturalis (IN) extract in periodontal regeneration by examining the viability of human gingival fibroblast (HGF), human periodontal ligament fibroblast (HPLF), and human osteoblast (HO) cells, as well as the effect of this extract on the secretion of type 1 collagen, bone morphogenetic protein-2 (BMP-2), and axin-2 in these cells. Materials and Methods: In our study, IN extract was analyzed using high-performance liquid chromatography (HPLC). Different concentrations of 5, 10, and 15 μg/ ml were prepared and applied to HGF, HPLF, and HO cell lines for 24 hours, followed by viability assessment using the MTT assay. Subsequently, a concentration of 5 μg/ ml was applied to HPLF and HO cells to evaluate the expression levels of type 1 collagen, BMP-2, and axin-2 proteins at 24 hours using the ELISA analysis. SPSS software package was used for statistical evaluation of the results. Results: IN applied at the tested concentrations did not show a significant effect on the viability of HGF, HPLF, and HO cells (p>0.05). The application of 5 μg/ ml IN to HPLF cells did not significantly affect the secretion of type 1 collagen and BMP-2 (p>0.05), but significantly increased the secretion of actin-2 (p=0.01). The application of 5 μg/ ml IN to HO cells significantly increased the secretion of type 1 collagen, BMP-2, and axin-2 (p=0.01, p=0.01, p=0.01). Conclusion: Within the limits of this research, the data obtained revealed that IN applied at different concentrations exhibited a similar effect that did not significantly impact the viability of HGF, HPLF, and HO cells. Additionally, IN may have positive effects on protein expression in HO cells, potentially through the activation of the Wnt signaling pathway. Further studies are needed where IN is applied at different concentrations and evaluated over varying follow-up periods.