靶向舌癌多功能纳米粒的制备和体外实验

doi:10.7518/hxkq.2018.03.002

华西口腔医学杂志 ›› 2018, Vol. 36 ›› Issue (3): 240-246.doi: 10.7518/hxkq.2018.03.002

靶向舌癌多功能纳米粒的制备和体外实验

任为¹(), 邱丽华^1,²(), 高志³, 李攀⁴, 赵欣⁵, 胡程晨¹

1.重庆医科大学附属口腔医院颌面外科,口腔疾病与生物医学重庆市重点实验室
2.重庆市高校市级口腔生物医学工程重点实验室,重庆 401147
3.重庆医科大学附属第二医院口腔科,重庆 400016
4.重庆医科大学超声影像学研究所,重庆 400010
5.重庆第二师范学院重庆市功能性食品协同创新中心,重庆 400067

收稿日期:2017-11-14 修回日期:2018-03-28 出版日期:2018-06-01 发布日期:2018-06-01
作者简介:
任为,硕士,E-mail：286525345@qq.com
基金资助:
重庆市社会民生科技创新专项项目（cstc2016shmszx000-10）;2016年重庆高校创新团队建设计划资助项目（CXTDG20160-2006）;重庆市高校市级口腔生物医学工程重点实验室资助项目[渝教科（2014）55号]

Preparation of multifunctional nanoparticles targeting tongue cancer and in vitro study

Wei Ren¹(), Lihua Qiu^1,²(), Zhi Gao³, Pan Li⁴, Xin Zhao⁵, Chengchen Hu¹

1. Dep. of Oral and Maxillofacial Surgery, Stomatological Hospital of Chongqing Medical University, Chongqing Key Laboratory of Oral Diseases and Biomedical Science, Chongqing 401147, China
2.Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China
3. Dept. of Stomatology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
4. Institute of Ultrasound Imaging, Chongqing Medical University, Chongqing 400010, China
5. Chongqing Collaborative Innovation Center for Functional Food, Chongqing University of Education, Chongqing 400067, China

Received:2017-11-14 Revised:2018-03-28 Online:2018-06-01 Published:2018-06-01
Supported by:
Chongqing Science and Technology Innovation Projects of Social Livelihood (cstc2016shmszx00010);;Program for Innovation Team Building at Institutions of Higher Education in Chongqing in 2016(CXTDG201602006);;Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education [(2014)55].

摘要/Abstract

摘要：

目的制备一种基质细胞衍生因子-1（SDF-1）修饰的载多西紫杉醇（DOC）包裹吲哚菁绿（ICG）和液态氟碳全氟己烷（PFH）靶向多功能纳米粒（DOC-SINPs）,检测其性质。方法双乳化法制备纳米粒,硫醚键连接SDF-1,得到靶向纳米粒。Malvern粒径仪检测其粒径及表面电位,激光扫描共聚焦显微镜观察SDF-1在纳米粒表面的连接情况,高效液相色谱法（HPLC）测定其DOC包封率（ELC）和载药量（DLC）及体外释放规律,热成像仪记录其体外光热特性,光声仪及超声诊断仪观察其体外显像,流式细胞仪评估其体外靶向能力,CCK-8法检查其对舌癌SCC-15细胞活力的影响。结果靶向多功能纳米粒形态规则,呈球状。平均粒径（502.88±17.92） nm,平均电位（-11.5±3.15） mV。平均ELC为54.12%±1.74%,平均DLC为1.08 mg·mL^-1。体外药物释放规律呈初期爆发性释放,接着是持续缓慢释放。其光热特性呈浓度相关,浓度越高温度越高。纳米粒可检测到明显光声信号和超声信号。体外靶向连接率89.99%。细胞活性实验结果表明,SINPs组在各个浓度下细胞活性无明显差异（P>0.05）。在浓度为150、200 μg·mL^-1时,DOC-SINPs无论有无激光辐照均较SINPs有明显的细胞活性抑制（P<0.05）。结论成功制备了集诊断与治疗一体的多功能纳米粒,具有超声/光声双模成像,同时具备化疗和光热治疗的抗肿瘤作用。

关键词: 聚乳酸-羟基乙酸共聚物, 吲哚菁绿, 靶向治疗, 舌癌, 分子成像

Abstract:

Objective This study aims to prepare docetaxel (DOC)-loaded multifunctional nanoparticles containing indo-cyanine green (ICG) and perfluorohexane (PFH) as targeted drug delivery system, which is supplemented with stromal cell-derived factor-1 (SDF-1), and characterize their properties. Methods Multifunctional nanoparticles were prepared by using the double emulsion method. SDF-1 was covalently conjugated to the surface of the nanoparticles through thioe-ther bonding. Their particle size, distribution, and surface potential were determined with the Malvern measuring in-strument. The conjugation of SDF-1 was evaluated by confocal laser scanning microscope. Encapsulation efficiency (ELC), drug loading capacity (DLC), and release regularity of the nanoparticles were determined by high-performance liquid chro-matography (HPLC). In vitro photothermal property was recorded by a thermal imager. The in vitro imaging capacity was observed by a photoacoustic instrument and an ultrasonic diagnostic apparatus. Targeting capability was assessed by flow cytometry. The cell activity on SCC-15 cells was checked by CCK-8 method. Results The targeted multifunctional nano-particles showed regularly sphericity. The diameter was (502.88±17.92) nm. The zeta potential was (-11.5±3.15) mV. ELC was 54.12%±1.74%. DLC was 1.08 mg·mL^-1. In vitro drug release was initially fast and subsequently slow. The photothermal characteristics were related to the concentration; the higher the concentration, the higher the temperature. Nanoparticles could detect significant photoacoustic and ultrasound signals. The in vitro targeting rate was 89.99%. No significant differences of cell viability in the SINPs groups were observed at each concentration (P>0.05). The inhibition effect of DOC-SINPs was stronger than that of SINPs whether or not in the presence of laser irradiation among the groups of 150 and 200 μg·mL^-1 (P<0.05). Conclusion Multifunctional nanoparticles for diagnosis and treatment were successfully prepared and displayed dual-mode ultrasound/photoacoustic imaging and antitumor effects of chemotherapy and photothermal therapy.

Key words: poly (lactic-co-glycolic acid), indocyanine green, targeted therapy, tongue carcinoma, molecular imaging

中图分类号:

R739.8

任为, 邱丽华, 高志, 李攀, 赵欣, 胡程晨. 靶向舌癌多功能纳米粒的制备和体外实验[J]. 华西口腔医学杂志, 2018, 36(3): 240-246.

Wei Ren, Lihua Qiu, Zhi Gao, Pan Li, Xin Zhao, Chengchen Hu. Preparation of multifunctional nanoparticles targeting tongue cancer and in vitro study[J]. West China Journal of Stomatology, 2018, 36(3): 240-246.

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参考文献 27

[1]	陈新, 徐文华, 周健, 等. 口腔鳞状细胞癌现状[J]. 口腔医学, 2017, 37(5): 462-465.
	Chen X, Xu WH, Zhou J, et al.Current situation of oral squa-mous cell carcinoma[J]. Stomatology, 2017, 37(5): 462-465.
[2]	Moore SR, Johnson NW, Pierce AM, et al.The epidemio-logy of mouth cancer: a review of global incidence[J]. Oral Dis, 2000, 6(2): 65-74.
[3]	Riemann M, Knipfer C, Rohde M, et al.Oral squamous cell carcinoma of the tongue: prospective and objective speech evaluation of patients undergoing surgical therapy[J]. Head Neck, 2016, 38(7): 993-1001.
[4]	Chang CC, Yang YJ, Li YJ, et al.MicroRNA-17/20a func-tions to inhibit cell migration and can be used a prognostic marker in oral squamous cell carcinoma[J]. Oral Oncol, 2013, 49(9): 923-931.
[5]	Kaneda MM, Caruthers S, Lanza GM, et al.Perfluorocarbon nanoemulsions for quantitative molecular imaging and tar-geted therapeutics[J]. Ann Biomed Eng, 2009, 37(10): 1922-1933.
[6]	Rapoport N, Christensen DA, Kennedy AM, et al.Cavitation properties of block copolymer stabilized phase-shift nanoe-mulsions used as drug carriers[J]. Ultrasound Med Biol, 2010, 36(3): 419-429.
[7]	徐芬芬, 王志刚, 李攀, 等. 载10-羟基喜树碱液态氟碳纳米粒制备及其表征、体外增强超声显像效果观察[J]. 中国医学影像技术, 2014, 30(8): 1127-1130.
	Xu FF, Wang ZG, Li P, et al.Preparation of 10-HCPT loaded perfluoropentane nanoparticles and observation its characte-ristic, enhancement effect for ultrasound imaging in vitro[J]. Chin J Med Imaging Technol, 2014, 30(8): 1127-1130.
[8]	刘建新, 尚婷婷, 徐芬芬, 等. 叶酸靶向液态氟碳脂质纳米粒的制备及其超声分子显像卵巢癌实验[J]. 中国医学影像技术, 2016, 32(12): 1843-1847.
	Liu JX, Shang TT, Xu FF, et al.Preparation of folic acid targeted liquid fluorocarbon lipid nanoparticles and appli-cation in ultrasound molecular imaging of ovarian cancer[J]. Chin J Med Imaging Technol, 2016, 32(12): 1843-1847.
[9]	Lowe KC.Engineering blood: synthetic substitutes from fluorinated compounds[J]. Tissue Eng, 2003, 9(3): 389-399.
[10]	Zheng C, Zheng M, Gong P, et al.Indocyanine green-loaded biodegradable tumor targeting nanoprobes for in vitro and in vivo imaging[J]. Biomaterials, 2012, 33(22): 5603-5609.
[11]	Jori G, Spikes JD.Photothermal sensitizers: possible use in tumor therapy[J]. J Photochem Photobiol B, 1990, 6(1/2): 93-101.
[12]	Jain RA.The manufacturing techniques of various drug loaded biodegradable poly(lactide-co-glycolide) (PLGA) devices[J]. Biomaterials, 2000, 21(23): 2475-2490.
[13]	Delilbasi CB, Okura M, Iida S, et al.Investigation of CXCR4 in squamous cell carcinoma of the tongue[J]. Oral Oncol, 2004, 40(2): 154-157.
[14]	张骁, 束梅英, 张韬. 多西紫杉醇用于癌症治疗的临床研究进展[J]. 中国制药信息, 2005, 21(12): 3-9.
	Zhang X, Su MY, Zhang T.Clinical research progress of docetaxel for cancer treatment[J]. Chin Pharma Inf, 2005, 21(12): 3-9.
[15]	Li J, Feng L, Fan L, et al.Targeting the brain with PEG-PLGA nanoparticles modified with phage-displayed peptides[J]. Biomaterials, 2011, 32(21): 4943-4950.
[16]	Lu W, Zhang Y, Tan YZ, et al.Cationic albumin-conjugated pegylated nanoparticles as novel Drug carrier for brain deli-very[J]. J Control Release, 2005, 107(3): 428-448.
[17]	Habash RW, Bansal R, Krewski D, et al.Thermal therapy, part 1: an introduction to thermal therapy[J]. Crit Rev Bio-med Eng, 2006, 34(6): 459-489.
[18]	Campbell RB.Tumor physiology and delivery of nanophar-maceuticals[J]. Anticancer Agents Med Chem, 2006, 6(6): 503-512.
[19]	Iyer AK, Khaled G, Fang J, et al.Exploiting the enhanced permeability and retention effect for tumor targeting[J]. Drug Discov Today, 2006, 11(17/18): 812-818.
[20]	Mu X, Zhang F, Kong C, et al.EGFR-targeted delivery of DOX-loaded Fe3O4 polydopamine multifunctional nano-composites for MRI and antitumor chemo-photothermal therapy[J]. Int J Nanomedicine, 2017, 12: 2899-2911.
[21]	Nguyen HT, Phung CD, Thapa RK, et al.Multifunctional nanoparticles as somatostatin receptor-targeting delivery system of polyaniline and methotrexate for combined chemo-photothermal therapy[J]. Acta Biomater, 2018, 68: 154-167.
[22]	张晓燕, 平其能. 多西紫杉醇白蛋白纳米粒的制备及体外评价[J]. 药学进展, 2008, 32(5): 223-228.
	Zhang XY, Ping QN.Preparation and in vitro evaluation of bovine serum albumin nanoparticles containing docetaxel[J]. Progress Pharma Sci, 2008, 32(5): 223-228.
[23]	Meng W, Xue S, Chen Y.The role of CXCL12 in tumor microenvironment[J]. Gene, 2018, 641: 105-110.
[24]	Chittasupho C, Lirdprapamongkol K, Kewsuwan P, et al.Targeted delivery of doxorubicin to A549 lung cancer cells by CXCR4 antagonist conjugated PLGA nanoparticles[J]. Eur J Pharm Biopharm, 2014, 88(2): 529-538.
[25]	Mei L, Liu Y, Zhang Q, et al.Enhanced antitumor and anti-metastasis efficiency via combined treatment with CXCR4 antagonist and liposomal doxorubicin[J]. J Control Release, 2014, 196: 324-331.
[26]	Janssens R, Struyf S, Proost P.The unique structural and functional features of CXCL12[J]. Cell Mol Immunol, 2017. doi: 10.1038/cmi.2017.107.
[27]	Li X, Qin F, Yang L, et al.Sulfatide-containing lipid per-fluorooctylbromide nanoparticles as paclitaxel vehicles targeting breast carcinoma[J]. Int J Nanomedicine, 2014, 9: 3971-3985.

靶向舌癌多功能纳米粒的制备和体外实验

Preparation of multifunctional nanoparticles targeting tongue cancer and in vitro study

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