納米生物學(xué)及其應(yīng)用-2

亓立峰浙江加州國際納米技術(shù)研究院:?前言：納米微粒概述Part

I:納米藥物與靶向設(shè)計(jì)Part

II:腫瘤納米分子影像與實(shí)時(shí)治療Part

III:納米探針:?前景展望刖aWhat

is

Nano?zcl^iPhycoerythrin丿I

nmIOO

nmPolymer

NPQuantum

dots,

magnetic

NPFigi.

A

schemetic

of

nanoscale

materialI

III10

pm

IOO

pmAtom

Small

DyeMolrcuiesA

uorescen

t

Col

Joida

HBacterin

eProtons

Cold刖aPolymer

NPDegradablePolymericmembrane某種物質(zhì)或元素納米級別粒子:nano

Zinic;SiO2?nano

gold,CdSe/ZnS,Fe2O3?PolymericmatrixnanospherenanocapsuleOily

or

aqueous

core寛C0I刖日納米微粒制備方法化學(xué)法：（1）沉淀法/透析法把沉淀劑加入到鹽溶液中反應(yīng)后，（將沉淀熱處理）得到納米材料。其特點(diǎn)簡單易行，但純度低，顆粒半徑大，聚合體納米粒子、氧化物。（2）微乳液法/乳液擴(kuò)散法兩種互不相溶的溶劑在表面活性劑的作用下形成乳液，在微泡中經(jīng)成核、

聚結(jié)、團(tuán)聚、（熱處理）后得納米粒子。其特點(diǎn)粒子的單分散和界面性好，聚合體納米粒子、II?VI族半導(dǎo)體納米粒子多用此法制備zcl^i納米粒子表征方法測量方法

測量功能離心沉降法

等效直徑適用的尺寸范圍使用的主:要儀器高速離心機(jī)，分光光度計(jì)或暗場法光學(xué)系統(tǒng)BET吸附裝置或重量法裝置喇曼光譜儀X射線衍射儀＞25nm氣體吸附法（容量法或重量法）光散射法X射線衍射峰寬法比表面積尺寸：約1?10nm比袤面積：0.1?1000m2/g〉約3]101約＜500nm

常用于＜5Onm平均直徑晶粒平均尺寸小角度X射線散射線晶粒平均尺寸

約＜100nmX射線衍射儀電子成像法（TEM,測量粒徑尺寸直接觀察粒了形貌并約＞2nni透射電子顯微鏡掃描隧道顯微鏡法（STM）形貌與尺寸寬范圍掃描隧道顯微鏡粒徑分析儀熒光光譜儀粒徑分析熒光光譜法粒徑分布熒光圖譜zcl^ipFairt

Ih納釆葯物與腫瘤霜向治辨IP攻dfe仙Enhanced

permeability

and

retention

(EPR)penetrate

through

the

leaky

pathologicalThese

newly

formed

tumor

vessels

are

usuallyabnormal

in

form

and

architecture.

They

arepoorly-aligned

defective

endothelial

cells

wiwide

fenestrations,

lacking

a

smooth

musclelayer,

or

innervation

with

a

wider

lumen,

andimpaired

functional

receptors

for,Furthermore,

tumor

tissuesusually

lack

effective

lymphatic

drainage.

Althese

factors

will

lead

to

abnormal

molecularand

fluid

transport

dynamics

especially

formacromolecular

drugs.

Namely,

thiseffect.

Long-circulating

drug

carriers

(1),

phenomenon

was

coined

""enhancedpermeability

and

retention

(EPR)-effect/z

ofvasculature

(2)z

into

the

tumor

interstitium

macromolecules

and

lipids

in

solid

tumors.(3),

and

degrade

there,

releasing

a

freedrug

(4)

and

creating

its

high

localconcentration.10.1208/aaDSj0902015寛C0I納米藥物與腫瘤靶向治療Nanoparticles

:

clinicaladministration

of

anticancerdrugs.Controlled

and

targeteddelivery

of

the

drugReduced

systemic

sideeffects

Facilitatedextravasation

into

the

tumorcells

(EPR

effect:

enhancedpermeability

and

retention)High

capability

to

crossvarious

physiological

barrierszcl^iChitosan的13

mVMiMitarntfta-CNP50

mVIMCNP?CuFig.3

Size

and

zeta

potential

of

CNP

and

CNP-CuL.

Qi,

et

al,

Colloids

Surfaces

A:

Physicochem

Eng

Aspects

2004;

251:

183190

;

Carbohydrate

Research

2004;

2693-2700zcl^iTable

1.

Cytotoxic

activity

of

chitosan,

CNP

(mean

particle

size

=

40

nm),

andCNP-Cu

against

different

cell

linesPanel

of

cell

lineCytotoxicity

(IC50,gg/mL)Cell

lineChitosan

CNP

CNP-CuLiverL-02na

na

naColon

cancerCalo3201200(±2

28(

+

3) 14(

+

2)7)Gastric

cancerBGC8231200(

+

35) 21(

+

2)

8(

+

1)Liver

cancerBEL74021200(

+

22) 15(

+

2)

6(

+

1)Values

are

means

of

three

experiments,

standard

deviation

is

given

inparentheses

(=not

active).L.

Qi,

et

al,

Bioorganic

&

Medicinal

Chemistry

Letters

2005;

15:

1357-1399Rh

1232040720

001RI1123*nc*>

"M

:M1;

7.30%家於L

*s-o

-u.，Q，RM23Fig.5

CNP-induced

changes

of

MMP

and

zeta

potential

of

membrane.

A:

Loss

of

MMP

and

zeta

potential;

Histogram

of

untreated

cells

(B)

and

cells

treated

with

100

pg/mL

CNP

(C)CNP

could

neutralize

the

negative

surface

charge

of

BEL7402

cells

soas

todamage

the

cell

membrane.

The

strongdissipation

in

MMP

suggests

apossible

disruption

of

mitochondrial

membrane

after

treated

with

CNP.L.

Qi,

et

al,

Eur.

Cancer

20Q7;

43:184-93zcl^iTable

2

Effects

ol

CNP

on

fatty

acid

composition

oimembrane

phosphoric

lipid

from

BEL7402

cells1.4apvO.Ol

vs

ControlIncreased

ROS

(Reactive

oxygen

species)production

and

LPO

degree,

decreasedunsaturated

fatty

acid

indicated

that

CNPexerted

membrane

penetration

activity

byinduction

of

LPO100oo25

5075CNP

(pg/mL)Fig.

6

Lipid

peroxidation

measured

asthiobarbituricacid

reactive

substances(TBARs)

production

of

BEL7402

cellsnontreated

or

treated

for

24

h

with

variousdoses

of

CNP.Fatty

acidControl

(|ig/mg)CNP

group

(Fig/mg)C.4:029.93±2.0130.29

+

2.21以:0216.5±13.

73215.51

+

5.41Ci&oC】8:l255.61

+

14.

86142.62±8.

52252.11

±10.

99C18:2226.22±8.

29195.78

±4.

54

a。20：412.97+1.

462142??o1s81

?o七61oMoi

)es

a

v

m

lzcl^iFige

7

Effects

of

administration

routes

onantitumor

efficacy

of

CNP

againstSarcoma-180

subcutaneous

tumorformation

and

growth

in

ICR

mice012345678Days

after

tivatinentControlChitosan40

nmCNP2.40+0.442.38

+0.481.96±0.85-2.3

±0.991.54

±0.56

2.95

±0.461.78

±0.97

2.24±0.41b70

nm

CNP100

nm

CNPCispeflfaetcitin勺u(yù)f

ehcmuther241.13±0.22b

621.21±0.29b

591.92±0.46b

350.56±0.13b

81120Table

4

Effect

ol

CNP

with

different

particle

sizeon

BEL7402

tumor

cell

growth

in

nude

mice

byoral

administration

(1

mg/kg/day)Groups

BW

TW

(g)

TIR

(%)

increase(g)2

0

Q

o886421(

L

U

M

ou

-n

wAJ

o

u

m

HImaging:

cancer

diagnostics,

staging, radiation

planning,

and

evaluation

of therapy.Standard

clinical

imaging

modalities: CT,

MRI,

ultrasound,

limitation

in detection

(

<

0.5

cm),

and distinguishing

between

benign

and cancerous

tumors.Molecular

imaging:

Multifunctional nanoparticles:

MNP,

QDs,

gold

NP, drug

delivery

(siRNA,

DNA,

drug), nanothermotherapy,

photodynamic therapy.What

is

Qdotszcl^iWavelength

(nm|Size-tunable

light

emissionPhotostableSimultaneous

excitation

of

QDsLarge

Stokes

shift0

一400500

600

700Wavelength

(nm)rrrn)A)—

SC5匚—

Oueausajon-I.2

1.8.6.4

.21ooooe5

?:>c<

Bq

§

q

m

w

N

l

l

eEJONzcl^iFigure

13.

Schematic

illustration

of

a

multifunctional

quantum

dotcoated

with

amphiphilic

polymer.QD-siRFMAEndocytosisFig.

14.

Schematic

diagram

showing

the

steps

of

siRNA-QD

in

membrane

binding,

cellular

enendosomal

escape,

capturing

by

RNA

binding

proteins,

loading

to

RNA-induced

silencing

co(RISC),

and

target

degradation.zcl^iAmphipol:

Poly

(maleicanhydride-alt-l-decene)modified

withDimethylaminopropylamine

(PMAD-N3)(d)Fig.

16.

Schematic

drawing

of

the

hybrid

structure

of

QD

and

amphipol

forsiRNA

delivery

and

real-time

imaging

in

live

cells.Qi

and

ACS

nanof

2008,

2(7),

1403-1410,2008zcl^isiRNA/QDMolar

Ratko:

50

40

30

20

10

1

0Fig.

17

QD

loading

capacity

and

protection

of

siRNA

molecules

againstnuclease

degradation

determined

by

gel

electrophoresis-Qi

and

Gao,

ACS

nano,

2008,

2(7),

1403-1410zcl^iFig.

18

Gene

silencing

eniciency

of

siRNA

targeting

Her-2

using

QDPMAL

comparedwith

the

classic

transfection

agents,

Lipofectamine

and

PEI.Qi

and

Gao,

ACS

nano,

2008,

2(7),

1403-14103寛C0I納米分子造影劑產(chǎn)品3TP的癌癥診斷技術(shù)，基于MRI的診斷乳腺癌、前列腺癌，區(qū)分惡性/良性，〉5mm瘤塊。主要以札/氧化鐵為組分核磁納米造影劑Palatin技術(shù)公司2007年獲批準(zhǔn)新型造影劑neutrospec,neutrospec含有一個(gè)以得為標(biāo)記的抗cdl5單克隆抗思可以選擇性的結(jié)合于參與免疫應(yīng)答的嗜中性粒細(xì)胞。

neutrospec被注射入血后，可與感染部位存在的嗜屮性

粒細(xì)胞相結(jié)合，使這些細(xì)胞被放射性示蹤劑所標(biāo)記。zcl^iPartlll:納米探針Chemical

&

Functional

Genomics

Nanotechnology^.Modulating

signaling

pathways

&

probing

biological

behaviors

為Chemical

libraries

Self-renewalFig.22

Modulating

signaling

pathways

and

probing

biological

interactions

withnanotopographyzcl^iFig.

25.

Cytotoxicity

of

CNCdetermined

by

detection

ofLDH

release

from

BJ-5a

cellscocultured

with

extracts

ofCNC

for

72

h.Chitosan3

h3dlidn

t

rc

o5

0

4

0

3

0

2

0

1

0Fige

26Actin

localization

in

spreading

cells

with

time3.2納米纖維轉(zhuǎn)染DNAzcEiiFig.

27.

Human

embryonic

kidney

cell

line

(HEK

293T)

cultured

onSiNWs

(c)

with

and

(d)

without

PEI

treatment

prior

to

GFP

plasmiddeposition.

Yang

et.

al.

J.

AM.

CHEM.

SOC.

2007,129,

7228-72293.3靶分子監(jiān)測納米探針zcl^iWavelength

(nm)Fig.

28.

Well-dispersed

antibody-nanoshell

conjugates

in

the

absence

of

analytepossess

a

well-defined

extinction

peak

in

the

near-IR.

In

the

presence

of

thecomplementary

analyte,

multiple

nanoshells

bind

to

the

analyte,

causing

agglutinationand

acorresponding

reduction

in

the

extinction

peak.Methods

in

Molecular

Biology?

?

303NANOBIOTECHNOLOGYPROTOCOLS3.3納米金熒光檢測核昔酸序列寛C0IFig.

29.

Signal

detection

of

oligonucleotide-derivatized

NBCshybridized

with

complementary

Cy5

oligonucleotide.Methods

in

Molecular

Biology?

?

303NANOBIOTECHNOLOGYPROTOCOLSzcl^i凱鳳應(yīng)齣釆籍維願(yuàn)于現(xiàn)瞰B測Fig.

30.

Flow

chamber

setup

for

hybridization

and

visualization

of

fluorescence

signals

on

thenanoarray.

Optical

microscope

image

of

an

optical

fiber-bundle

based

nanoarray

containingindivi