利用静电自组装法制备了壳聚糖/siRNA纳米粒,建立了SYBR Gold核酸染料法测定siRNA复合率(CR),并系统性考察了n(N)/n(P)比,壳聚糖分子量(Mw)、脱乙酰度(DD)及制备环境pH值、缓冲液浓度(Bc)对纳米粒siRNA复合率、粒径及表面电位的影响规律.结果表明,siRNA复合率由n(N)/n(P) 0.5时的(18.6±4.0)%快速增大至n(N)/n(P)为2时的(92.9±2.0)%,但随n(N)/n(P)增大复合率不再升高;n(N)/n(P)<2时,难以形成稳定纳米粒,复合物表面电位为负,n(N)/n(P)由5升至150时,纳米粒粒径由(179.3±5.7) nm增大至(235.5±8.3) nm,表面电位由(17.2±1.3) mV升高至(23.6±0.9) mV.另外,n(N)/n(P) 150时,CR随DD升高而升高,随Bc升高而降低;纳米粒粒径随Mw增大而增大,随DD及缓冲液浓度(Bc)升高而减小;纳米粒表面电位随DD升高而升高,随pH值及Bc升高而降低.n(N)/n(P)比对纳米粒特性及siRNA复合率影响最为显著,调整n(N)/n(P)、Mw、DD、pH值及离子强度可得到siRNA复合率在90%以上,粒径100~300 nm,表面电位15~25 mV的纳米粒.
参考文献
[1] | Peacock, H.;Kannan, A.;Beal, P.A.;Burrows, C.J..Chemical modification of siRNA bases to probe and enhance RNA interference[J].The Journal of Organic Chemistry,201118(18):7295-7300. |
[2] | Tiziana Musacchio;Vladimir P Torchilin.siRNA delivery: from basics to therapeutic applications.[J].Frontiers in bioscience: a journal and virtual library,2013P1/393(P1/393):58-79. |
[3] | M. Dash;F.Chiellini;R.M. Ottenbrite.Chitosan-A versatile semi-synthetic polymer in biomedical applications[J].Progress in Polymer Science,20118(8):981-1014. |
[4] | Ragelle, H.;Vandermeulen, G.;Préat, V..Chitosan-based siRNA delivery systems[J].Journal of Controlled Release: Official Journal of the Controlled Release Society,20131(1):207-218. |
[5] | Garcia-Fuentes, M.;Alonso, M.J..Chitosan-based drug nanocarriers: Where do we stand?[J].Journal of Controlled Release: Official Journal of the Controlled Release Society,20122(2):496-504. |
[6] | Yuan X;Shah BA;Kotadia NK;Li J;Gu H;Wu Z.The development and mechanism studies of cationic chitosan-modified biodegradable PLGA nanoparticles for efficient siRNA drug delivery.[J].Pharmaceutical research,20107(7):1285-1295. |
[7] | Kong,F.;Liu,G.;Sun,B.;Zhou,S.;Zuo,A.;Zhao,R.;Liang,D..Phosphorylatable short peptide conjugated low molecular weight chitosan for efficient siRNA delivery and target gene silencing[J].International Journal of Pharmaceutics,20121/2(1/2):445-453. |
[8] | Nadithe, Venkatareddy;Liu, Runhui;Killinger, Bryan A.;Movassaghian, Sara;Kim, Na Hyung;Moszczynska, Anna B.;Masters, Kristyn S.;Gellman, Samuel H.;Merkel, Olivia. M..Screening Nylon-3 Polymers, a New Class of Cationic Amphiphiles, for siRNA Delivery[J].Molecular pharmaceutics,20152(2):362-374. |
[9] | Zheng,M.;Librizzi,D.;Kili?,A.;Liu,Y.;Renz,H.;Merkel,O.M.;Kissel,T..Enhancing in vivo circulation and siRNA delivery with biodegradable polyethylenimine-graft-polycaprolactone-block-poly(ethylene glycol) copolymers[J].Biomaterials,201227(27):6551-6558. |
[10] | Kevin Buyens;Martin Meyer;Ernst Wagner;Joseph Demeester;Stefaan C. De Smedt;Niek N. Sanders.Monitoring the disassembly of siRNA polyplexes in serum is crucial for predicting their biological efficacy[J].Journal of Controlled Release: Official Journal of the Controlled Release Society,20101(1):38-41. |
[11] | Alabi, Christopher A.;Sahay, Gaurav;Langer, Robert;Anderson, Daniel G..Development of siRNA-probes for studying intracellular trafficking of siRNA nanoparticles[J].Integrative Biology: quantitative biosciences from nano to macro,20131(1):224-230. |
[12] | Tore-Geir lversen;Tore Skotland;Kirsten Sandvig.Endocytosis and intracellular transport of nanoparticles: Present knowledge and need for future studies[J].Nano Today,20112(2):176-185. |
[13] | Wei, A.;Mehtala, J.G.;Patri, A.K..Challenges and opportunities in the advancement of nanomedicines[J].Journal of Controlled Release: Official Journal of the Controlled Release Society,20122(2):236-246. |
[14] | Rosemary Kanasty;Joseph Robert Dorkin;Arturo Vegas.Delivery materials for siRNA therapeutics[J].Nature materials,201311(11):967-977. |
[15] | Kwon, I.K.;Lee, S.C.;Han, B.;Park, K..Analysis on the current status of targeted drug delivery to tumors[J].Journal of Controlled Release: Official Journal of the Controlled Release Society,20122(2):108-114. |
[16] | Katas H;Alpar HO.Development and characterisation of chitosan nanoparticles for siRNA delivery[J].Journal of Controlled Release: Official Journal of the Controlled Release Society,20062(2):216-225. |
[17] | Liu X;Howard KA;Dong M;Andersen MO;Rahbek UL;Johnsen MG;Hansen OC;Besenbacher F;Kjems J.The influence of polymeric properties on chitosan/siRNA nanoparticle formulation and gene silencing.[J].Biomaterials,20076(6):1280-1288. |
[18] | Luo, Y.;Zhai, X.;Ma, C.;Sun, P.;Fu, Z.;Liu, W.;Xu, J..An inhalable β_2-adrenoceptor ligand-directed guanidinylated chitosan carrier for targeted delivery of siRNA to lung[J].Journal of Controlled Release: Official Journal of the Controlled Release Society,20121(1):28-36. |
[19] | Gordon A. Morris;Jonathan Castile;Alan Smith.Macromolecular conformation of chitosan in dilute solution: A new global hydrodynamic approach[J].Carbohydrate Polymers: Scientific and Technological Aspects of Industrially Important Polysaccharides,20094(4):616-621. |
[20] | 高群;王国建;李文涛.壳聚糖在稀溶液中的分子构象及其影响因素[J].化学通报(网络版),2009(1):1-6. |
[21] | 宋巍;陈元维;史国齐;张小华;余喜讯;万昌秀.不同脱乙酰度壳聚糖的制备及结构性能的研究[J].功能材料,2007(10):1705-1708. |
[22] | Xu SL;Dong MD;Liu XD;Howard KA;Kjems J;Besenbacher F.Direct force measurements between siRNA and chitosan molecules using force spectroscopy[J].Biophysical Journal,20073(3):952-959. |
[23] | 张威;夏文水.离子强度对壳聚糖分子链构象的影响[J].食品科学,2014(9):34-37. |
上一张
下一张
上一张
下一张
计量
- 下载量()
- 访问量()
文章评分
- 您的评分:
-
10%
-
20%
-
30%
-
40%
-
50%