Synthesis of PEGylation hydroxyapatite drug delivery system and its dual channels fluorescence imaging

doi:10.11949/0438-1157.20200780

Abstract

Abstract:

PEGylated drug delivery systems (DDSs) can overcome the side effects of traditional chemotherapy by enhancing drug permeability and retention (EPR) effects. In this work, DOX@HAP (hydroxyapatite) was initially fabricated via the coprecipitation and hydrothermal method, further functionalized with Cy (cyanine) by coupling reaction of APTES and then introduced hydrophilic PEG chains by using copper(I)-catalyzed alkyne–azide cycloaddition reaction. Physicochemical properties including the morphology, particle size and phase composition, were characterized by TEM, SEM, particle size analyzer, FTIR, XPS and XRD. The encapsulation efficiency and drug release profile of DOX@HAP-Cy-PEG were analyzed by UV-Vis spectrophotometry. Furthermore, the cellular uptake of DOX@HAP-Cy-PEG nanoparticles in Hela and HepG2 cells was monitored by the dual channels fluorescence imaging of DOX and Cy. The results showed that DOX@HAP-Cy-PEG nanoparticles could be used to real-time monitor the dynamic distribution of DDSs in Hela and HepG2 cells by dual channels.

Key words: hydroxyapatite, drug delivery system, cyanine, dual channels, fluorescence imaging

摘要：

PEG化的药物递送系统（DDSs）可以通过增强药物的渗透性和滞留性（EPR）效应克服传统化疗的副作用。利用共沉淀法和水热法制备纳米粒子DOX@HAP，进一步通过偶联反应修饰菁染料（Cy），通过铜（I）催化的炔-叠氮化物环加成反应修饰PEG链，构建了纳米制剂DOX@HAP-Cy-PEG。通过透射电子显微镜（TEM）、扫描电子显微镜（SEM）、粒度分析仪、傅里叶红外光谱仪（FTIR）、X射线光电子能谱仪（XPS）和X射线衍射仪（XRD）对该纳米载药体系的形貌、粒径、物相组成进行表征分析。利用紫外-可见（UV-Vis）分光光度法测定了该纳米材料的药物负载量以及体外药物释放曲线。进一步，利用DOX和Cy双通道荧光成像，监测DDSs在Hela和HepG2细胞中的摄取行为。表明DOX@HAP-Cy-PEG纳米载药体系有望作为一种新型的治疗与示踪一体化的抗癌纳米制剂。

关键词: 羟基磷灰石, 载药体系, 菁染料, 双通道, 荧光成像

CLC Number:

TQ 34

Xiaojing LI, Wen SUN, Yao KANG, Jiangli FAN, Xiaojun PENG. Synthesis of PEGylation hydroxyapatite drug delivery system and its dual channels fluorescence imaging[J]. CIESC Journal, 2020, 71(10): 4808-4819.

李晓静, 孙文, 康垚, 樊江莉, 彭孝军. PEG化羟基磷灰石纳米体系的制备及双通道荧光成像[J]. 化工学报, 2020, 71(10): 4808-4819.

Figures/Tables 14

Fig.1 Reaction equation of cyanine dyes and PEG alkynyl

Fig.2 TEM (a) and SEM (b) images of DOX@HAP. DLS analysis (c), zeta potential (d) and PDI values (e) of DOX@HAP，DOX@HAP-Cy and DOX@HAP-Cy-PEG

Fig.3 XPS (a) and XRD (b) patterns of DOX@HAP. FTIR spectra (c) of HAP，DOX@HAP，DOX@HAP-Cy and DOX@HAP-Cy-PEG

Fig.4 The absorption spectra (a) and fluorescence spectra (b) of DOX，Cy and DOX@HAP-Cy-PEG

Fig.5 The absorption spectra of the supernatant liquid of DOX@HAP treated with PBS at pH 3.0 for 24 h(a). The linear relationship between UV absorption (λab = 500 nm) and concentrations of DOX: (A = 16.27C + 0.0476)(b). Release profiles of DOX from DOX@HAP-Cy-PEG in PBS buffer with different pH (5.0,6.5,7.4) in 24 h(c)

Fig.6 Dose-dependent cytotoxicity of Hela and HepG2 treated with HAP (a)，HAP-Cy-PEG (b) and DOX@HAP-Cy-PEG (c) at various concentrations

Fig.7 CLSM images of Hela and HepG2 cells incubated with DOX@HAP-Cy-PEG for 4 h by dual channels fluorescence imaging

Fig.A1 Synthetic route of PEG-alkyne

Fig.A2 Synthetic route of DOX@HAP-Cy-PEG

Fig.A3 ESI-MS spectrum of compound 4

Fig.A4 1H NMR spectrum of compound 4

Fig.A5 ESI-MS spectrum of Cy

Fig.A6 1H NMR spectrum of Cy

Table A1 XPS elemental analysis of DOX@HAP

Element	Peak binding energy/eV	Content/%(atom)
P	132.75	8.96
Ca	346.88	14.6
C	284.49	27.75
O	530.89	48.7

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