纳米材料的可控合成、功能化及其在肿瘤诊疗中应用研究
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摘要
大规模高效地制备大小均一、形貌可控的纳米材料一直是研究的热点问题，是推动纳米科学和纳米技术发展的关键。接踵而至的纳米材料的表面功能化推动了纳米技术在生物医学领域中的广泛应用。在新兴的纳米生物医学领域中，将具有先进功能的纳米材料及具有智能响应特性的纳米结构用于疾病的诊断和治疗研究，不仅能为疾病的早期诊断以及发生发展过程提供更直观的证据，而且还有望实现影像介导的药物递送和治疗、影像指导的手术切除和实时监控的治疗应答等。
本论文内容主要分为两个部分：第一部分是关于纳米材料的两种可控合成方法的开发及其机理研究；第二部分是关于纳米材料的功能化及其在肿瘤诊疗中的应用研究。详述如下：
1.利用液-液两相法可控合成了金属纳米材料和上转换材料。（1）利用室温甲苯（油相）和水（水相）两相体系，成功地合成了单分散的银、铜、镍和铁等金属纳米颗粒，并对其形成机理进行了讨论。（2）利用油酸-离子液体两相体系，成功地合成了一系列不同形貌的尺寸小于25nm的稀土氟化物上转换发光纳米晶体，包括LaF_3、GdF_3、YF_3、YbF_3、NaGdF_4、NaYF_4和NaYbF_4等，并对其形成机理进行了讨论。
2.利用牛血清白蛋白（BSA）作为辅助，调控硫族半导体纳米材料的成核与生长，成功地合成了硒化银、硒化镉、硒化铅和硒化铜等纳米材料，并对其生长过程和机理进行了深入的研究，在此基础上对其形成机理提出了假说。
3.设计和制备了光敏剂偶联的磁性纳米探针。该纳米探针在外加磁场作用下具有磁导向性可以实现肿瘤的磁靶向；在交变磁场作用下，能快速升温具有热疗作用；在激光照射下，展现出光动力学治疗作用；同时该纳米探针还具有核磁共振成像和近红外（NIR）成像的功能，最终有望实现磁靶向药物递送和肿瘤磁荧光成像监控的光动力学和磁致热双模式治疗。
4.设计和制备了叶酸偶联的二氧化硅包覆的金纳米棒纳米探针。该纳米探针具有肿瘤靶向性、对放射治疗具有增敏作用、具有显著的光热治疗作用，同时，还可以作为X-ray/CT成像的对比剂。该纳米探针有望实现对肿瘤的靶向X-ray/CT成像监控的放射增敏和光热双模式治疗。
5.设计和制备了叶酸偶联的氧化石墨烯纳米载体。通过疏水相互作用和π-π共轭作用成功地将光敏剂药物二氢卟吩（Ce6）装载到氧化石墨烯表面上，该纳米载体能显著地提高Ce6在肿瘤细胞内聚集的效率，在激光照射下具有显著的光动力学治疗作用。该纳米探针有望实现对肿瘤的靶向成像和光动力学治疗。
6.设计和制备了光敏剂偶联的碳点纳米探针。利用荧光共振能量转移（FRET）原理，碳点能显著提高光敏剂NIR成像的能力。该纳米探针具有很好的稳定性，无细胞毒性，良好的生物相容性，能高效地在肿瘤部位蓄积，最终成功实现了对肿瘤组织的增强荧光成像监控的光动力学治疗。
The development of efficient methods for the controlled synthesis of nanocrystalswith monodispersity, stability, and predictable morphology is one of the key goals in theadvancement of nanoscience and nanotechnology. Afterwards, the surfacefunctionalization of nanomaterials promotes the extensive applications of nanotechnologyin biomedical field. In the burgeoning nano-bio-medicine field, use of advancednanomaterials and smart stimuli-responsive nanostructures for the diagnosis and treatmentof disease can provide the direct evidence to early diagnosis, occurrence and developmentprogresses of disease, and also enables online imaging of drug for the detection of disease,image-guided drug delivery and treatments, guidance of surgical resection, and monitoringof treatment response.
This dissertation mainly includes (1) two kinds of controlled synthesis methods ofnanomaterials and their mechanism investigation.(2) surface functionalization ofnanomaterials for tumor theranostic applications. The main contributions are as follows:
1. Controlled synthesis of metallic and upconversion rare earth fluoride nanocrystals vialiquid-liquid two-phase system.(1) Monodisperse metallic nanocrystals with a narrow sizedistribution such as Ag, Cu, Ni and Fe nanoparticles were synthesized in the toluene-watertwo-phase system at room temperature. A possible mechanism for the formation ofmonodisperse nanocrystals is also discussed.(2) Highly uniform and monodisperse REF_3and NaREF_4(RE=La, Gd, Yb and Y) nanocrystals with small size (<25nm) anddifferent morphologies were synthesized in the oleic acid (OA)/ionic liquids (ILs)two-phase system. A possible mechanism for the formation of monodisperse nanocrystalsis also discussed.
2. Bovine serum albumin (BSA)-conjugated MxSey(M=Ag, Cd, Pb, Cu) nanomaterialswith different shapes and sizes were synthesized in water at room temperature by aprotein-directed, solution-phase, green synthetic method. A possible mechanism for theformation of semiconductor chalcogenides in the BSA solution is discussed.
3. Design and preparation of photosensitizer-conjugated magnetic nanoparticles. Thisnanoprobe can offer magnetically guided drug delivery under external magnetic field,induce hyperthermia under an alternating magnetic field, and display remarkablephotodynamic efficacy upon laser irradiation. Our results indicated that the synthesizedmultifunctional nanoprobe is potential for magnetically guided drug delivery,simultaneous magnetofluorescent imaging-guided dual-modal photodynamic therapy(PDT) and hyperthermia.
4. Design and preparation of folic acid-conjugated silica-modified gold nanorods. Thisnanoprobe show highly selective targeting, enhanced radiation therapy (RT) andphoto-thermal therapy (PTT) effects on MGC803gastric cancer cells, and also exhibitedstrong X-ray attenuation for in vivo X-ray and computed tomography (CT) imaging. Thesynthesized nanoprobe is a good candidate with excellent imaging and targeting ability forX-ray/CT imaging-guided targeting dual-modal enhanced RT and PTT.
5. Design and preparation of folic acid-conjugated graphene oxide (GO). Thephotosensitizer (PS) Chlorin e6(Ce6) was effectively loaded into the system viahydrophobic interactions and π-π stacking. The nanocarriers can significantly increase theaccumulation of Ce6in tumor cells and lead to a remarkable photodynamic efficacy onMGC803cells upon irradiation. These suggested that folic acid-conjugated GO loadedCe6had great potential as effective drug delivery system in targeting PDT.
6. Design and preparation of chlorin e6-conjugated C-dots (C-dots-Ce6). C-dots canenhance the fluorescence of Ce6on C-dots-Ce6by the F rster (fluorescence) resonanceenergy transfer (FRET). The C-dots-Ce6owns good stability, noncytotoxicity, goodbiocompatibility, enhanced photosensitizer fluorescence detection and remarkablephotodynamic efficacy upon irradiation. The synthesized nanoprobe is effective forsimultaneous enhanced-fluorescence imaging-guided PDT.
本论文内容主要分为两个部分：第一部分是关于纳米材料的两种可控合成方法的开发及其机理研究；第二部分是关于纳米材料的功能化及其在肿瘤诊疗中的应用研究。详述如下：
1.利用液-液两相法可控合成了金属纳米材料和上转换材料。（1）利用室温甲苯（油相）和水（水相）两相体系，成功地合成了单分散的银、铜、镍和铁等金属纳米颗粒，并对其形成机理进行了讨论。（2）利用油酸-离子液体两相体系，成功地合成了一系列不同形貌的尺寸小于25nm的稀土氟化物上转换发光纳米晶体，包括LaF_3、GdF_3、YF_3、YbF_3、NaGdF_4、NaYF_4和NaYbF_4等，并对其形成机理进行了讨论。
2.利用牛血清白蛋白（BSA）作为辅助，调控硫族半导体纳米材料的成核与生长，成功地合成了硒化银、硒化镉、硒化铅和硒化铜等纳米材料，并对其生长过程和机理进行了深入的研究，在此基础上对其形成机理提出了假说。
3.设计和制备了光敏剂偶联的磁性纳米探针。该纳米探针在外加磁场作用下具有磁导向性可以实现肿瘤的磁靶向；在交变磁场作用下，能快速升温具有热疗作用；在激光照射下，展现出光动力学治疗作用；同时该纳米探针还具有核磁共振成像和近红外（NIR）成像的功能，最终有望实现磁靶向药物递送和肿瘤磁荧光成像监控的光动力学和磁致热双模式治疗。
4.设计和制备了叶酸偶联的二氧化硅包覆的金纳米棒纳米探针。该纳米探针具有肿瘤靶向性、对放射治疗具有增敏作用、具有显著的光热治疗作用，同时，还可以作为X-ray/CT成像的对比剂。该纳米探针有望实现对肿瘤的靶向X-ray/CT成像监控的放射增敏和光热双模式治疗。
5.设计和制备了叶酸偶联的氧化石墨烯纳米载体。通过疏水相互作用和π-π共轭作用成功地将光敏剂药物二氢卟吩（Ce6）装载到氧化石墨烯表面上，该纳米载体能显著地提高Ce6在肿瘤细胞内聚集的效率，在激光照射下具有显著的光动力学治疗作用。该纳米探针有望实现对肿瘤的靶向成像和光动力学治疗。
6.设计和制备了光敏剂偶联的碳点纳米探针。利用荧光共振能量转移（FRET）原理，碳点能显著提高光敏剂NIR成像的能力。该纳米探针具有很好的稳定性，无细胞毒性，良好的生物相容性，能高效地在肿瘤部位蓄积，最终成功实现了对肿瘤组织的增强荧光成像监控的光动力学治疗。
The development of efficient methods for the controlled synthesis of nanocrystalswith monodispersity, stability, and predictable morphology is one of the key goals in theadvancement of nanoscience and nanotechnology. Afterwards, the surfacefunctionalization of nanomaterials promotes the extensive applications of nanotechnologyin biomedical field. In the burgeoning nano-bio-medicine field, use of advancednanomaterials and smart stimuli-responsive nanostructures for the diagnosis and treatmentof disease can provide the direct evidence to early diagnosis, occurrence and developmentprogresses of disease, and also enables online imaging of drug for the detection of disease,image-guided drug delivery and treatments, guidance of surgical resection, and monitoringof treatment response.
This dissertation mainly includes (1) two kinds of controlled synthesis methods ofnanomaterials and their mechanism investigation.(2) surface functionalization ofnanomaterials for tumor theranostic applications. The main contributions are as follows:
1. Controlled synthesis of metallic and upconversion rare earth fluoride nanocrystals vialiquid-liquid two-phase system.(1) Monodisperse metallic nanocrystals with a narrow sizedistribution such as Ag, Cu, Ni and Fe nanoparticles were synthesized in the toluene-watertwo-phase system at room temperature. A possible mechanism for the formation ofmonodisperse nanocrystals is also discussed.(2) Highly uniform and monodisperse REF_3and NaREF_4(RE=La, Gd, Yb and Y) nanocrystals with small size (<25nm) anddifferent morphologies were synthesized in the oleic acid (OA)/ionic liquids (ILs)two-phase system. A possible mechanism for the formation of monodisperse nanocrystalsis also discussed.
2. Bovine serum albumin (BSA)-conjugated MxSey(M=Ag, Cd, Pb, Cu) nanomaterialswith different shapes and sizes were synthesized in water at room temperature by aprotein-directed, solution-phase, green synthetic method. A possible mechanism for theformation of semiconductor chalcogenides in the BSA solution is discussed.
3. Design and preparation of photosensitizer-conjugated magnetic nanoparticles. Thisnanoprobe can offer magnetically guided drug delivery under external magnetic field,induce hyperthermia under an alternating magnetic field, and display remarkablephotodynamic efficacy upon laser irradiation. Our results indicated that the synthesizedmultifunctional nanoprobe is potential for magnetically guided drug delivery,simultaneous magnetofluorescent imaging-guided dual-modal photodynamic therapy(PDT) and hyperthermia.
4. Design and preparation of folic acid-conjugated silica-modified gold nanorods. Thisnanoprobe show highly selective targeting, enhanced radiation therapy (RT) andphoto-thermal therapy (PTT) effects on MGC803gastric cancer cells, and also exhibitedstrong X-ray attenuation for in vivo X-ray and computed tomography (CT) imaging. Thesynthesized nanoprobe is a good candidate with excellent imaging and targeting ability forX-ray/CT imaging-guided targeting dual-modal enhanced RT and PTT.
5. Design and preparation of folic acid-conjugated graphene oxide (GO). Thephotosensitizer (PS) Chlorin e6(Ce6) was effectively loaded into the system viahydrophobic interactions and π-π stacking. The nanocarriers can significantly increase theaccumulation of Ce6in tumor cells and lead to a remarkable photodynamic efficacy onMGC803cells upon irradiation. These suggested that folic acid-conjugated GO loadedCe6had great potential as effective drug delivery system in targeting PDT.
6. Design and preparation of chlorin e6-conjugated C-dots (C-dots-Ce6). C-dots canenhance the fluorescence of Ce6on C-dots-Ce6by the F rster (fluorescence) resonanceenergy transfer (FRET). The C-dots-Ce6owns good stability, noncytotoxicity, goodbiocompatibility, enhanced photosensitizer fluorescence detection and remarkablephotodynamic efficacy upon irradiation. The synthesized nanoprobe is effective forsimultaneous enhanced-fluorescence imaging-guided PDT.
引文
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