Selective cytotoxicity of Ferulic Acid-Loaded ZIF-8 nanoparticles and free Ferulic Acid on ovarian carcinoma and dental pulp stem cells

Abstract

Purpose: To overcome the rapid oxidation and low stability of Ferulic Acid (FA), this study designed and characterized FA-loaded Zeolitic imidazol framework -8 (FA-ZIF-8) nanoparticles, evaluating their selective cytotoxic, anti-proliferative, anti-migratory, and apoptotic effects on OVCAR-3 and SKOV-3 ovarian cancer cells alongside healthy primary dental pulp stem cells (DPSCs). Materials and methods: FA-ZIF-8 nanoparticles were synthesized and characterized. OVCAR-3, SKOV-3, and DPSC cohorts were treated with varying concentrations of FA (5 μM–1 mM) and FA-ZIF-8 (25–500 μg/mL) for up to 72 h. Cell viability and 48-h half-maximal inhibitory concentrations IC50 were quantified using the XTT assay. Transcriptional and translational expressions of apoptotic markers were analyzed via qRT-PCR and immunocytochemistry and TUNEL analysis. Long-term survival and lateral cell motility were monitored via coloni and scratch wound healing assays. Results: FA exhibited a non-toxic profile, actively stimulating the proliferation of both cancer lines and DPSCs. FA-ZIF-8 inverted this response, yielding 48-h IC50 values of 308.2098 μg/mL for OVCAR-3 and 336.3022 μg/mL for SKOV-3. FA-ZIF-8 upregulated Bax expression and Caspase-3 levels while suppressing Bcl-2, driving a significant increase in the Bax/Bcl-2 (p=0.0024). These results were confirmed by an increase in TUNEL-positive apoptotic cell. Furthermore, FA-ZIF-8 inhibited colony formation and significantly reduced lateral migration indices, particularly in SKOV-3 lines. Conclusion: FA acts as a proliferative ligand, its strategic formulation into FA-ZIF-8 nanoparticles enables a, "Trojan Horse" effect, delivering selective cytotoxicity, anti-metastatic suppression, pH-responsive apoptotic induction to epithelial ovarian cancer cells.

Keywords

• Zeolitic imidazol framework 8
• ovarian cancer
• ferulic acid
• apoptosis
• wound healing

Ethical Statement

This study was approved by Pamukkale University Ethic Committee for Non-Invasive Clinical Trials (approved date: 13.03.2026, and approved number: E-60116787-020-845567).

Ferulik asit içeren ZIF-8 nanopartiküllerinin ve serbest ferulik asidin ovaryum karsinomu ve diş pulpa kök hücreleri üzerindeki seçici sitotoksisitesi

Abstract

Amaç: Düşük stabilitesi nedeniyle klinik kullanımı sınırlı olan ferulik asidin (FA) hücre içi taşınımını iyileştirmek amacıyla pH'a duyarlı zeolitik imidazolat çerçeve-8 (ZIF-8) gibi nano-metal-organik çerçeveler kullanılmış ve FA-ZIF-8 nanopartikülleri tasarlanarak bu sistemin over kanseri hatlarında seçici sitotoksik, anti-migratif ve apoptotik etkileri sağlıklı dental pulpa kök hücreleri (DPSCs) kontrolü eşliğinde in vitro olarak değerlendirilmiştir. Yöntemler: FA-ZIF-8 nanopartikülleri sentezlenmiş ve karakterize edilmiştir. OVCAR-3, SKOV-3 ve DPSCs, 72 saate kadar serbest FA (5 μM–1 mM) ve FA-ZIF-8'in (25–500 μg/mL) artan konsantrasyonlarına maruz bırakılmıştır. IC50 değerleri XTT analizi ile belirlenmiştir. Apoptotik belirteçler sırasıyla qRT-PCR, immünositokimya ve TUNEL analizi ile doğrulanmıştır. Uzun vadeli üreme yeteneği ve lateral hücre hareketliliği sırasıyla koloni ve yara iyileşmesi analizleri ile izlenmiştir. Bulgular: Tek başına uygulanan serbest FA, toksik olmayan bir profil sergilemiş; hem kanser hatlarının hem de DPSC'lerin proliferasyonunu aktif olarak uyarmıştır. FA-ZIF-8 bu yanıtı tersine çevirerek, OVCAR-3 için 48 saatlik IC50 değerini 308,2098 μg/mL ve SKOV-3 için 336,3022 μg/mL olarak belirledi. FA-ZIF-8, Bcl-2'yi baskılarken Bax ekspresyonunu ve Caspase-3 düzeylerini artırdı ve bu da Bax/Bcl-2 oranında anlamlı bir artışa yol açtı (p=0,0024). Bu sonuçlar, TUNEL pozitif apoptotik hücre sayılarındaki istatistiksel olarak anlamlı artışla onaylanmıştır. Ayrıca FA-ZIF-8, koloni oluşumunu kısıtlamış ve özellikle SKOV-3 hatlarında lateral göç indekslerini anlamlı olarak azaltmıştır. Sonuç: FA proliferatif bir ligand gibi davranırken, FA-ZIF-8 nanopartikülleri şeklindeki stratejik formülasyonunun bir "Truva Atı" etkisi yarattığını; epitelyal over kanseri hücrelerine seçici sitotoksisite, anti-metastatik baskılama ve pH'a duyarlı apoptotik, indüksiyon sağladığı görülmüştür.

Keywords

• Zeolitik imidazolat çerçeve-8
• over kanseri
• ferulik asit
• apoptosis
• yara iyileşmesi

Ethical Statement

Bu çalışma,Pamukkale Üniversitesi Non-İnvaziv Klinik Araştırmalar Etik Kurulu tarafından onaylanmıştır (onay tarihi: 13.03.2026, onay numarası: E-60116787-020-845567).

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