基于Zn酞菁的高效上转换纳米粒子：NIR光敏剂的光动力疗法突破

201 浏览量更新于2024-08-28 收藏 3.74MB PDF 举报

上转换纳米粒子-基于锌酞菁的纳米光敏剂用于光动力疗法（Upconversion Nanoparticles - ZnPhthalocyanine-Based Nanophotosensitizer for Photodynamic Therapy）是当前光动力疗法领域的一项关键研究进展。传统的光动力疗法（PDT）依赖于对可见光的吸收，但上转换技术通过近红外（NIR）光激发，允许在较低的辐射剂量下实现高效的单线态氧（1O2）生成，这是治疗癌症等疾病的重要机制。ZnPc，即锌酞菁，作为一种光敏剂，其在NIR光下的吸收性能被用来增强上转换过程。近期的研究焦点在于如何设计和制备高效的上转换纳米光敏剂，如UCNPs-ZnPc。这种新型纳米材料由NaYF4：Yb3+，Er3+上转换纳米颗粒（UCNP）和Zn（II）-酞菁（ZnPc）光敏剂通过共价组装的方式结合。共价组装技术显著减少了纳米粒子间的距离，提高了能量转移效率，从而提高光致产率（O-1）。 UCNPs-ZnPc的实验结果显示，在体内实验中，仅需低剂量的980 nm辐射（351 J/cm²，约0.39 W/cm²），配合15分钟的短时间照射，就能在小鼠肝脏肿瘤模型中实现图像引导的光动力治疗，并实现了大约80.1%的肿瘤抑制效果。这一成就表明，即使在较低的辐射剂量下，上转换纳米光敏剂也能展现出卓越的治疗效果。然而，安全性也是关注的重点。组织学分析并未发现注入UCNPs-ZnPc后的心脏、肺、肾脏、肝脏或脾脏出现病理变化和炎症反应，这证明了该疗法的潜在优点。这些研究不仅推动了上转换纳米光疗的发展，也为未来的临床应用提供了可能，尤其是在那些难以到达且对传统光疗有抵抗性的肿瘤部位。总结来说，上转换纳米粒子结合锌酞菁作为光敏剂的研究展示了在光动力疗法中的巨大潜力，特别是在降低辐射剂量的同时保持高效治疗效果。共价组装技术的应用是关键，它优化了能量传输并确保了纳米结构的稳定性和生物相容性。随着这种技术的不断优化和深入研究，有望为未来的癌症治疗开辟新的途径。

ﬂuorophore products, and the excess ﬂuorescamine can be hydrolyzed into non-

ﬂuorescent products very fast. By measuring the emission band centered at

470 nm (

¼ 390 nm), the presence of amino group can be validated.

2.6. Surface modiﬁcation of the nanoparticles with FA and PEG

To covalently conjugate 2, 9, 16, 23-tetracarboxylic Zinc phthalocyanine and folic

acid on UCNPs, 5 ml of dimethylformamide solution containing 0.8 mg of 2, 9, 16, 23-

tetracarboxylic Zinc phthalocyanine, 0.5 mg of folic acid, 1 mg of 1-ethyl-3- (3-

dimethylaminopropyl) carbodiimide and 1 mg of N-hydroxy-succinimide were

incubated at room temperature for 2 h, and then 10 mg of amino-functionalized

NaYF

:Yb

3þ

,Er

3þ

upconversion nanoparticles was added into the solution and

stirred vigorously for 24 h. The mixture was then centrifuged at 10,000 rpm for

10 min three times to spin down the nanoparticles. The supernatant was withdrawn

carefully and the particles resuspended in DMSO and ethanol mixture solution

(DMSO: ethanol ¼ 4:1) for further used. To reduce the undesired toxicity of nano-

particles to normal tissue, the surface of UCNPs-ZnPc/FA were coated with PEG. 4 ml

of DMSO and ethanol mixture solution containing 1 mg PEG-SC and 1 mg UCNPs-

ZnPc/FA was shaken by shaking table. The mixture was then centrifuged at

10,000 rpm for 10 min to spin down the nanoparticles. The supernatant was

withdrawn carefully and the particles resuspended in PBS (PH ¼ 7.4). The washing

process was repeated twice.

2.7. To assess the NIR exposure effect on the cells

Hela cells that over express folate receptors and A549 that have a low expression

of folate receptors were purchased from First Bethune Hospital, University of Jilin.

HeLa cells were grown in a 96-well cell-culture plate at 10

e10

per well and then

incubated for 24 h at 37



C under 5% CO

. A power adjustable 980 nm ﬁber laser with

maximal output power of 30 W (n-LIGHT Corporation) was collimated and

employed as area light source to irradiate the 96-well plate. After 10 min exposure of

980 nm light at different power density (0, 0.39, 0.85, 1.23, 1.61, 1.99 W/cm

), the

cells were allowed to incubate for an additional 48 h. To assess the effect of exposure

to NIR laser on the cells, the cell viability was measured by 3-(4, 5-dimethylthiazol-

2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay.

2.8. Targeted cancer cell imaging and therapy

Hela cells cultured in folate-free RPMI-1640 medium was considered as the

positive group, while HeLa cells cultured in normal RPMI-1640 medium and A549

cells which were cultured in folate-free RPMI-1640 medium were considered as a

negative group. All the mediums were supplemented with 10% fetal bovine serum,

100

g/mL penicillin, and 100

g/mL streptomycin. Cells were cultivated in medium

at 37



C in a humidiﬁed 95% air and 5% carbon dioxide (CO

) atmosphere. The

cytotoxicity was measured using a standard MTT assay. HeLa cells were grown in a

96-well cell-culture plate at 10

e10

per well and then incubated for 24 h at 37



under 5% CO

. After 24 h culturing, UCNPs-ZnPc/FA nanoconjugates were added to

the culture medium at different concentrations, with ﬁve parallel wells for each

concentration (0, 100, 200, 400, 800

g/mL). The standard MTT assay was carried out

to determine the cell viabilities relative to the untreated cells. For the FA targeted

PDT experiment, the cells were allowed to incubate with different concentration

UCNPs-ZnPc/FA (0, 100, 200, 400, 800

g/mL) for another 24 h at 37



C and then

washed twice with PBS before being exposed to NIR irradiation. A power adjustable

980 nm ﬁber laser with maximal output power of 30 W was collimated and

employed as area light source to irradiate the 96-well plate. After 10 min exposure of

980 nm light at 0.39 W/cm

, the cells were allowed to incubate for an additional

48 h. To assess the effect of exposure to NIR laser on the cells with and without the

nanoparticles, the cell viability was measured by MTT assay. Typically, 10

L of MTT

solution (5 mg/mL MTT in PBS) was added to each well and incubated for 4 h at



C. After removing the medium, the wells were washed by PBS, and then the

intracellular formazan crystals were extracted into 100

l of DMSO. The absorbance

of cell lysate was recorded at 490 nm by a microtiter plate reader, and the cell

viability could be calculated from the average value of four parallel wells.

2.9. Upconversion luminescent imaging of cells

For the FA targeted imaging experiment, both positive and negative cells were

seeded in the confocal dishes at a concentration of 10

/ml. After 24 h of cell

attachment, both positive and negative cells were incubated with 200

g/mL UCNPs-

ZnPc/FA for 2 h at 37



C. Before imaging all cells were washed three times and ﬁxed

in 4% paraformaldehyde for 20 min at RT. Cells were washed twice with PBS three

times. The nuclei were then counterstained with 0.1

g/mL DAPI for 10 min at RT

followed by twice washing with PBS for three times. Upconversion ﬂuorescence

imaging were then performed using a Nikon Inverted Microscope Eclipse Ti-U Main

Body (Nikon, Tokyo, Japan) equipped with C2-SHS Scanner and Controller under

excitation of lower power density (0.19 W/cm

2.10. In vivo PDT

Female C57/6J mice (20 g, 6e 8 weeks old) used in this study were purchased

from First Bethune Hospital, University of Jilin. All experiments were carried out in

compliance with the animal management. The Hepa1-6 tumor model was estab-

lished by subcutaneously inoculating Hepa1-6 cells (3  10

) into the upper axillary

fossa in the mice (n ¼ 6). The mice were investigated when the tumor grew to a

diameter of 4e6 mm. 100

l saline or UCNPs-ZnPc/FA (10 mg/ml) was intratumorally

injected into each Hepa1-6 tumor-bearing mouse. The mice were randomly assigned

into four groups treated with different injections, as follows: (1) group 1 received

only intratumoral injection of the saline (the control group, n ¼ 6); (2) group 2

received intratumoral injection of the saline with NIR light irradiation (n ¼ 6); (3)

group 3 received intratumoral injection of the UCNPs-ZnPc/FA (n ¼ 6); (4) group 4

received intratumoral injection of the UCNPs-ZnPc/FA with NIR light irradiation

(n ¼ 6). The tumors were irradiated with a 980 nm laser light (0.39 W cm

2

) for

15 min. To avoid any tissue damage by heating, the laser treatment was done with

3 min interval for every 3 min of light exposure. After treatment, the tumor volume

was calculated as length  (width)

 1/2 with a caliper over 2 weeks. The body

weight of each mouse was monitored every other day over 2 weeks. Relative tumor

volume, relative body weight and inhibition ratio were deﬁned as follows: Relative

tumor volume ¼ V/V

, V

and V stand for the tumor volume on the initial day and on

the day of measurement, respectively. Relative body weight ¼ W/W

, W

and W are

the body weight of mouse on the initial day and on the day of measurement,

respectively. Inhibition ratio ¼ (Vc  Vt)/Vc  100%, Vt and Vc represent the average

tumor volume for the control group and treatment group, respectively.

2.11. Histology examination

Histology analysis was performed at the 14th day after treatment. The organs

(heart, liver, spleen,lungand kidney) and tumor tissuesof the mice in the control group

and treatment group were isolated from the mice, ﬁxed with 10% neutral buffered

formalin and embedded in parafﬁn. The sliced organs and tumor tissues (8 mm) were

stained with Hematoxylin and Eosin (H&E) and examined by a microscope.

3. Results and discussion

3.1. Synthesis of UCNPs and covalently constructing UCNPs-ZnPc/FA

nanophotosensitizer

The NaYF

: 25% Yb

3þ

, 0.2% Er

3þ

oleic acid coated UCNPs were

used as the donor, instead of 20% Yb

3þ

used generally in the current

studies to increase the upconversion luminescence (UCL) at 660 nm

(Fig. S1) and their morphologies and phase purities were analyzed

by TEM and XRD, respectively, as shown in Fig. 1 and Fig. S2. Good

Fig. 1. TEM image of NaYF

:Yb

3þ

,Er

3þ

UCNPs (a) before phase transfer and (b) after phase transfer.

L. Xia et al. / Biomaterials 35 (2014) 4146e41564148

剩余10页未读，继续阅读

weixin_38708105

粉丝: 9
资源: 865

基于Zn酞菁的高效上转换纳米粒子：NIR光敏剂的光动力疗法突破

不同长径比金纳米双锥对光敏剂荧光和光动力疗法效果的增强

锌酞菁体外光动力学疗法体外抗肿瘤细胞量效关系

论文研究 - 基于卟啉和酞菁衍生物-染料敏化太阳能电池的增感剂的共价无机-有机杂化分子框架的组装

配位溶解沉淀法制备纳米Y-酞菁氧钛 (2014年)

电子功用-基于酞菁钴标记电化学检测有机小分子的方法

酞菁类光敏剂单重态氧量子产额的激光诱导ESR研究

大数据-算法-手性卟啉酞菁的设计合成有机聚集及非线性光学性质研究.pdf

论文研究 - 过渡金属酞菁掺杂的玻璃离聚物水泥的结构表征

物联网-智慧传输-基于双异质诱导的超薄酞菁锌薄膜NO2传感器的研究.pdf

包含（8-喹啉氧基-5-磺酸）酞菁钴和氧化石墨烯的层膜增强的静电自组装层的反向饱和吸收

最新资源