산화아연 나노물질과 식품첨가물 용제의 상호작용 연구 = Interactions between ZnO nanoparticles and food additive solvents

다국어 초록 (Multilingual Abstract)

Zinc Oxide(ZnO) is utilized as a food additive to increase the nutritional value of zinc(Zn), an essential trace element, and acts as Zn fortifiter. The direct addition of ZnO to processed foods as a food additive can lead to interactions between ZnO nanoparticles (NPs) and the food matrix, such as carbohydrates, proteins, and fats. With the advancement of nanotechnology, ZnO can be produced as a nano-sized material with a high surface area-to-volume ratio, exhibiting greater reactivity and a wider range of reactions compared to bulk-sized materials. Therefore, interactions between ZnO NPs and food ingredients or additives must be considered when evaluating the potential toxicity and biological responses of ZnO directly added to food. While most studies to date have focused on interactions between nanomaterials and the food matrix, little information is available on the interactions of ZnO with food additives.
This study aimed to investigate the effects of interactions between ZnO NPs and three representative additive solvents (methanol, glycerin, and propylene glycol) on their toxicity, physicochemical properties, and biological responses. After exposure to these solvents, the hydrodynamic diameter, zeta potential, and solubility of ZnO NPs were measured, and their crystalline phase was analyzed to evaluate interaction-induced changes in physicochemical properties. To understand the relationship between these changes and biological responses, human intestinal epithelial cell lines and an in vitro human intestinal transport model were used to assess the effects on cytotoxicity, cellular uptake, and gastrointestinal absorption of ZnO NPs. Additionally, an ex vivo intestinal sac model using rat small intestine was employed to analyze the impact of solvent interactions on intestinal absorption and biological responses.
The results showed that the hydrodynamic diameter of ZnO NPs decreased significantly and their solubility increased when interacting with glycerin and propylene glycol, but not with methanol. However, these interactions did not alter the crystal structure of ZnO NPs. They induced increased cytotoxicity, including cell proliferation inhibition, membrane damage, and ROS generation, as well as enhanced cellular uptake and both passive and active transport of ZnO NPs. Taken together, these findings suggest that interactions between ZnO NPs and additive solvents can enhance their homogeneous dispersion and solubility, leading to a smaller hydrodynamic diameter and distinct biological responses compared to pure ZnO NPs.

목차 (Table of Contents)

• 1. 서론
• 2. 재료 및 방법
• 2.1. 나노물질과 시험 물질 준비 방법
• 2.2. ZnO NPs의 물리화학적 특성 분석
• 2.2.1. 구성 입자 크기 및 형태 분석
• 2.2.2. 수분산 입자 크기 및 표면 전하 측정
• 2.2.3. 결정 구조 분석
• 2.3. ZnO NPs의 정량 분석을 위한 표준곡선 작성
• 2.4. ICP-AES를 사용한 Zn 정량 분석
• 2.5. ZnO NPs의 용해도 분석
• 2.6. 세포주 및 배양 조건
• 2.7. 세포 성장 저해 실험(MTT assay)
• 2.8 세포막 손상 확인 실험(LDH release assay)
• 2.9. 세포 내 활성산소종 정량 분석 실험(ROS generation assay)
• 2.10. 장관 세포 내 ZnO NPs의 유입양상 비교
• 2.11. In vitro 위장관 흡수 메커니즘 비교
• 2.12. 막 투과성
• 2.13. 실험동물 및 사육 조건
• 2.14. Everted small intestinal sac을 통한 intesrinal absorption 비교
• 2.14.1. Everted small intestinal sac model 구축
• 2.14.2. ZnO NPs의 intesrinal absorption 비교
• 2.15. 통계 분석
• 3. 결과 및 고찰
• 3.1. ZnO NPs의 물리화학적 특성 분석
• 3.1.1. 구성 입자 크기 및 형태 분석
• 3.1.2. 수분산 입자 크기 및 표면 전하 측정
• 3.1.3. 결정 구조 분석
• 3.2. ZnO NPs의 정량 분석을 위한 표준곡선 작성
• 3.3. ZnO NPs의 용해도 분석
• 3.4. 세포 성장 저해 실험(MTT assay)
• 3.5. 세포막 손상 확인 실험(LDH release assay)
• 3.6. 세포 내 활성산소종 정량 분석 실험(ROS generation assay)
• 3.7. 장관 세포 내 ZnO NPs의 유입양상 비교
• 3.8. In vitro 위장관 흡수 메커니즘 비교
• 3.9. 막 투과성
• 3.10. ZnO NPs의 intesrinal absorption 비교
• 4. 결론
• 5. 참고문헌
• 6. 영문요약