Pre-assembled nanospheres in mucoadhesive microneedle patch for sustained release of triamcinolone in the treatment of oral submucous fibrosis  

湿态黏附微针贴片中预组装纳米球结构单元以缓释曲安奈德治疗口腔黏膜下纤维性变

作　　者：CHENG Xian YANG Yanqing HUANG Junming GUO Qiuyun ZHU Wei LONG Dingpei ZHOU Yueying FENG Hui WANG Jie LI Yusi ZHOU Jian LIU Yanping LIU Ousheng 程岘;杨艳青;黄俊铭;郭秋云;朱伟;龙定沛;周玥颖;冯慧;王洁;李宇思;周建;刘焱萍;刘欧胜(中南大学湘雅口腔医院/湘雅口腔医学院,湖南省数智化与个体化诊疗技术工程研究中心,湖南省3D打印口腔医疗工程技术研究中心,口腔颌面再生王松灵院士专家工作站,长沙410078;海南省人民医院(海南医学院附属海南医院)口腔科,海口570311;湖南大学机械与运载工程学院,国家高效磨削工程技术研究中心,长沙410082;西南大学资源昆虫高效养殖与利用全国重点实验室,重庆400715;首都医科大学附属北京口腔医院口腔涎腺疾病中心,全牙再生及口腔组织功能重建北京市重点实验室,口腔健康北京实验室,北京100050)

机构地区：[1]Xiangya Stomatological Hospital and Xiangya School of Stomatology,Central South University,Hunan Engineering Research Center for Oral Digital Intelligence and Personalized Medicine,Hunan 3D Printing Engineering Research Center of Oral Care,WANG Songling Academician Workstation for Oral-maxilofacial and Regenerative Medicine,Central South University,Changsha 410078 [2]Department of Stomatology,Hainan General Hospital(Affiliated Hainan Hospital of Hainan Medical University),Haikou 570311 [3]College of Mechanical and Vehicle Engineering,National Engineering Research Centre for High Efficiency Grinding,Hunan University,Changsha 410082 [4]State Key Laboratory of Resource Insects,Southwest University,Chongqing 400715 [5]Salivary Gland Disease Center,Affiliated Stomatological Hospital of Capital Medical University,Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction,Beijing Laboratory of Oral Health,Beijing 100050,China

出　　处：《中南大学学报（医学版）》2024年第8期1245-1260,共16页Journal of Central South University ：Medical Science

基　　金：supported by the National Key Research and Development Program(2022YFC2402900);the Key Research and Development Program of Hainan Province(ZDYF2024SHFZ128);the Science and Technology Innovation Program of Hunan Province(2022RC1213),China.

摘　　要：Objective:Drug-loaded mucoadhesive silk fibroin(SF)microneedle patch can overcome the limitations of low bioavailability and significant pain associated with traditional treatment methods,such as topical application or injection of triamcinolone for oral submucous fibrosis(OSF).However,these systems release the drug too quickly,failing to meet the clinical requirements.This study aims to construct a mucoadhesive SF microneedle patch pre-assembled with silk fibroin nanospheres(SFN)and explore its ability to sustain the release of triamcinolone in the treatment of OSF.Methods:SFN was pre-assembled via precipitation reaction and characterized by scanning electron microscope(SEM)for the morphology.The particle size andζ-potential were measured by dynamic light scattering(DLS).Triamcinolone was loaded onto SFN using a diffusional post-loading method.The effective loading of triamcinolone was confirmed using Fourier-transform infrared spectroscopy(FTIR).The concentration of unloaded triamcinolone was quantified by high-performance liquid chromatography.Drug encapsulation efficiency and loading capacity of SFN were then calculated to determine the optimal amount of drug loading.The SFN suspension was pre-mixed with SF solution to prepare the microneedle under-layer.The microneedle morphology was observed by SEM.Compression mechanical tests were performed to evaluate the fracture force of microneedles at different nanosphere contents(5%,10%,and 20%),determining the optimal pre-mixing ratio.Ex-vivo mouse oral mucosa permeation studies were performed to ascertain the insertion depth of the microneedles via histological sections.The adhesive top layer was synthesized using SF and tannic acid,with FTIR confirming its successful synthesis.Its viscoelasticity was characterized by a rheometer,and differential scanning calorimetry analyzed thermal properties.Tensile tests evaluated the interfacial bonding strength between the adhesive layer and microneedle base to ensure no detachment during use.Adhesion to wet oral mucosal tis目的:载药湿态黏附蚕丝蛋白(silk fibroin,SF)微针贴片能克服涂抹或注射曲安奈德等传统口腔黏膜下纤维性变(oral submucous fibrosis,OSF)治疗方式中存在的药物生物利用度低和注射疼痛明显的缺陷,但其释放药物速率过快,无法满足临床需要。本研究旨在构建预组装SF纳米球(silk fibroin nanosphere,SFN)结构单元的湿态黏附SF微针贴片,探究其缓释曲安奈德治疗OSF的能力。方法:通过沉淀反应法制备SFN,采用扫描电镜(scanning electron microscope,SEM)观察其形貌,动态光散射(dynamic light scattering,DLS)测量其粒径及表面ζ-电位;通过后载法加载曲安奈德,傅里叶变换红外光谱(Fourier-transform infrared spectroscopy,FTIR)验证药物是否成功加载,高效液相色谱测定未加载上的药物浓度以间接计算药物包封率和SFN的载药率,确定最佳载药浓度比。将SFN分散成悬液,与SF溶液预混后制备微针底层,采用SEM观察微针形貌及针尖形态,通过压缩力学测试检测不同SFN含量(5%、10%、20%)制备的微针的机械强度以确定最佳预混比例,随后将其应用于离体小鼠口腔黏膜后进行组织切片,确定其插入深度。制备蚕丝-丹宁酸湿态黏附顶层,通过FTIR验证其是否成功合成,流变仪表征其黏弹性,差示扫描量热法分析其热效应特性,拉伸力学测试测量其与微针底层之间的界面结合力以确保不发生界面脱离,进一步检测其与离体湿润口腔黏膜组织间的湿态黏附性,并与现有商业口腔贴片对比。使用前述优化后的条件制备双层湿态黏附微针贴片,与成纤维细胞共培养,通过细胞计数试剂盒-8(cell counting kit-8,CCK-8)、活死染色及鬼笔环肽染色检测其细胞相容性;通过释药实验检测湿态黏附微针贴片的药物缓释性能。结果:成功制备SFN(平均粒径46.25 nm),在曲安奈德与SFN质量比为0.5时,达到最大药物包封率[(63.88±1.09)%],此时SF载药率为(27.41±3.06)%。预�

关 键 词：oral disease oral mucosal disease oral submucous fibrosis mucoadhesive microneedle patch 

分 类 号：R781.5[医药卫生—口腔医学]