Conjugated Polymers for Biological and Biomedical Applications E-Book

• Herausgeber: Wiley-VCH Verlag GmbH & Co. KGaA
• Kategorie: Wissenschaft und neue Technologien
• Sprache: Englisch

Seitenzahl: 721

Veröffentlichungsjahr: 2018

Beschreibung

This first book to specifically focus on applications of conjugated polymers in the fields of biology and biomedicine covers materials science, physical principles, and nanotechnology.
The editor and authors, all pioneers and experts with extensive research experience in the field, firstly introduce the synthesis and optical properties of various conjugated polymers, highlighting how to make organic soluble polymers compatible with the aqueous environment. This is followed by the application of these materials in optical sensing and imaging as well as the emerging applications in image-guided therapy and in the treatment of neurodegenerative diseases.
The result is a consolidated overview for polymer chemists, materials scientists, biochemists, biotechnologists, and bioengineers.

Leseprobe

Table of Contents

Cover

Title page

Copyright

Preface

Chapter 1: Strategies to Bring Conjugated Polymers into Aqueous Media

1.1 Introduction

1.2 Synthesis of CPEs

1.3 Neutral WSCPs

1.4 Fabrication of CPNPs

1.5 Conclusion

References

Chapter 2: Direct Synthesis of Conjugated Polymer Nanoparticles

2.1 Introduction

2.2 Generation of CPNs

2.3 Conclusion

References

Chapter 3: Conjugated Polymer Nanoparticles and Semiconducting Polymer Dots for Molecular Sensing and

In Vivo

and Cellular Imaging

3.1 Introduction

3.2 Preparation, Characterization, and Functionalization

3.3 Molecular Sensing

3.4 Cellular Imaging

3.5 Conclusion

Acknowledgment

References

Chapter 4: Conjugated Polymers for

In Vivo

Fluorescence Imaging

4.1 Introduction

4.2

In Vivo

Fluorescence Imaging of Tumors

4.3 Stimuli‐Responsive Fluorescence Imaging

4.4

In Vivo

Fluorescence Cell Tracking

4.5 Two‐Photon Excited Brain Vascular Imaging

4.6 Dual‐Modality Imaging of Tumors

In Vivo

4.7 Other

In Vivo

Fluorescence Imaging Applications

4.8 Conclusions and Perspectives

References

Chapter 5: π‐Conjugated/Semiconducting Polymer Nanoparticles for Photoacoustic Imaging

5.1 Introduction

5.2 Mechanism of PA Imaging

5.3 SPNs for PA Imaging

5.4 Summary and Outlook

References

Chapter 6: Conjugated Polymers for Two‐Photon Live Cell Imaging

6.1 Introduction

6.2 Conjugated Polymers and CPNs as One‐Photon Excitation Imaging Contrast Agents

6.3 Conjugated Polymers as 2PEM Contrast Agents

6.4 Conjugated‐Polymer‐Based Nanoparticles (CPNs) as 2PEM Contrast Agents

6.5 Conclusions and Outlook

References

Chapter 7: Water‐Soluble Conjugated Polymers for Sensing and Imaging Applications

7.1 Introduction

7.2 Conjugated Polymers for Sensing

7.3 Imaging of Conjugated Polymers

7.4 Challenges and Outlook

References

Chapter 8: Conjugated Polymers for Gene Delivery

8.1 Introduction

8.2 Fundamental Properties of Conjugated Polymers

8.3 Intracellular Targeting, Cytotoxicity, and Biodegradability of Conjugated Polymers

8.4 Plasmid DNA (pDNA) Delivery

8.5 Small Interfering RNA (siRNA) Delivery

8.6 Conclusions and Outlook

References

Chapter 9: Conductive Polymer‐Based Functional Structures for Neural Therapeutic Applications

9.1 Introduction

9.2 Conductive Polymer‐Based Functional Structures

9.3 Synthesis and Functionalization of Conductive Polymer‐Based Functional Structures

9.4 Applications of Conductive Polymer‐Based Functional Structures for Neural Therapies

9.5 Summary and Outlook

References

Chapter 10: Conjugated Polymers for Photodynamic Therapy

10.1 Introduction

10.2 Conjugated Polymers as Photosensitizers

10.3 Applications of CP‐Based Photodynamic Therapy

10.4 Conclusions and Future Perspectives

References

Chapter 11: Conjugated Polymers for Near‐Infrared Photothermal Therapy of Cancer

11.1 Introduction

11.2 Conjugated Polymers for Cancer Photothermal Therapy

11.3 Imaging Guided Photothermal Therapy

11.4 Conjugated Polymers for Combination Cancer Treatment

11.5 Outlook and Perspectives

References

Chapter 12: Conjugated Polymers for Disease Diagnosis and Theranostics Medicine

12.1 Introduction

12.2 Disease Diagnostics via Conjugated Polymers

12.3 Conjugated Polymers for Cancer Theranostics

12.4 Studying Neurodegenerative Disorders

References

Chapter 13: Polymer‐Grafted Conjugated Polymers as Functional Biointerfaces

13.1 Introduction

13.2 Methods of Functionalizing CPs

13.3 CP‐Based Polymer Brushes as Biointerfaces: Rationale and Applications

13.4 Synthesis of CP‐Based Graft Copolymer Brushes

13.5 Conclusions and Outlook

References

Index

End User License Agreement

List of Tables

Chapter 11: Conjugated Polymers for Near‐Infrared Photothermal Therapy of Cancer

Table 11.1 A summary of various conjugated polymeric nanoparticles explored in photothermal therapy.

List of Illustrations

Chapter 1: Strategies to Bring Conjugated Polymers into Aqueous Media

Scheme 1.1 Illustration of typical structures of CPE (a), neutral WSCP (b), and CPNP (c). The red color represents the CP backbone.

Scheme 1.2 Polymerization methods most widely employed to construct conjugated backbones. Ar

1

and Ar

2

represent aromatic units.

Scheme 1.3 Representative strategies for synthesis of CPEs through incorporation of charges via direct polymerization (a) and postpolymerization (b) method.

Scheme 1.4 Synthesis of sulfonated polythiophenes

P2–P4

.

Scheme 1.5 Synthesis of sulfonated CPEs

P6

and

P7

with single‐bonded backbones through Suzuki polymerization method.

Scheme 1.6 Synthesis of sulfonated CPEs

P10–P13

with double‐bonded backbones.

Scheme 1.7 Synthesis of sulfonated CPEs

P14

and

P15

with triple‐bonded backbones through Sonogashira reaction.

Scheme 1.8 Synthesis of carboxylated polythiophenes

P17

,

P19

, and

P20

.

Scheme 1.9 Synthesis of carboxylated CPEs

P21

,

P24

, and

P26

with single‐bonded backbones.

Scheme 1.10 Synthesis of carboxylated CPEs

P28

and

P29

with double‐bonded backbones.

Scheme 1.11 Synthesis of carboxylated CPEs

P30

,

P32

, and

P33

with triple‐bonded backbones.

Scheme 1.12 Synthesis of phosphonated CPEs

P35

,

P36

, and

P38

.

Scheme 1.13 Synthesis of ammonium polythiophenes

P39

,

P41

, and

P44

.

Scheme 1.14 Synthesis of ammonium CPEs

P45

,

P47

,

P49

,

P51

,

P52

, and

P56

with single‐bonded backbones.

Scheme 1.15 Synthesis of ammonium CPEs

P57

,

P59

,

P61

, and

P63

with double‐bonded backbones.

Scheme 1.16 Synthesis of ammonium polyfluorenes

P64

and

P66

with triple‐bonded backbones.

Scheme 1.17 Synthesis of hyperbranched CPE

P68

.

Scheme 1.18 Synthesis of pyridinium CPEs

P69

,

P71

, and

P72

.

Scheme 1.19 Synthesis of cationic CPEs

P73

,

P74,

and

P76

.

Scheme 1.20 Synthesis of zwitterionic CPEs

P77

,

P79

, and

P81

.

Scheme 1.21 Synthesis of neutral WSCPs

P82

and

P85

.

Scheme 1.22 Synthesis of neutral WSCP

P87

.

Scheme 1.23 Synthesis of neutral WSCP

P91

with HPG brush.

Scheme 1.24 Representative chemical structures of CPs and amphiphilic materials used for CPNP preparation.

Scheme 1.25 Schematic illustration of the preparation of CPNPs using reprecipitation method.

Scheme 1.26 Schematic illustration of the preparation of CPNPs using miniemulsion method.

Scheme 1.27 Schematic illustration of the preparation of CPNPs using nanoprecipitation method.

Chapter 2: Direct Synthesis of Conjugated Polymer Nanoparticles

Figure 2.1 Common conjugated polymers, their derivatives, and polymerization types.

Figure 2.2 Schematic representations of the nanoprecipitation, miniemulsification, and self‐assembly methods.

Figure 2.3 Electron microscopy images of CPNs synthesized by nanoprecipitation: (a) MEH‐PPV CPNs. () (b, c) Polythiophene CPNs, the scale bars represent 200 nm. () (d) (F8BT) Conjugated polymer shell seeded onto silica coated gold nanorods. () (e) Rod‐like CPNs. ()

Figure 2.4 CPNs synthesized by miniemulsification: (a) transmission electron microscope image of poly(

p

‐phenylene) nanoparticles. () (b) Transmission electron micrograph of poly(phenylene–ethynylene) nanoparticles. The scale bar represents 500 nm, and (c) confocal microscope image (left) and bright field image (right) of a HeLa cell incubated with polyethylene glycolated phospholipid‐functionalized CPNs, the scale bar represents 10 µm. (.) (d) Scanning electron microscopy image of polyfluorene particles. The scale bar is 200 nm. Inset: dynamic light scattering profile of a poly(fluorene) particle dispersion. ()

Figure 2.5 Comparison of microfluidic processes: (a) miniemulsification and (b) obtained particles. () (c) Microfluidic nanoprecipitation and (d) obtained particles. ()

Figure 2.6 Schematic of the described direct particle preparation methods: (a) emulsion polymerization (b) polymerization in mini‐ and (c) microemulsion and (d) dispersion polymerization.

Figure 2.7 Nanoparticles synthesized by emulsion polymerization: (a) TEM image of fluorescent nanoparticles. () (b) SEM image of PEDOT particles. () (c) SEM image of PEODT nanoparticles deposited on a silicon substrate (left) and TEM image of a single PEDOT nanoparticle (right). (d)

In vivo

fluorescence image of a tumor bearing mice after injection of PEDOT nanoparticles. ()

Figure 2.8 TEM images of CPNs synthesized by polymerization in miniemulsion. (a) Polyfluorene/quantum dot hybrid nanoparticles. Inset displays an individual core/shell hybrid particle. The scale bars represent 50 nm, and (b) poly(fluorene) nanoparticles (the scale bar represents 100 nm). () (c) Poly(acetylene) nanoparticles synthesized by Glaser coupling, and (d) CPN dispersions with tunable emission colors. ()

Figure 2.9 Images of CPNs synthesized by dispersion polymerization: Scanning electron micrograph images of (a) monodisperse particles of F8BT, (b) ellipsoidal particles of stretched monodisperse F8BT particles, (c) bidisperse particles of F8BT, (d) monodisperse polyfluorene particles produced by Suzuki–Miyaura dispersion polymerization. () (e) Monodisperse poly(phenylene–vinylene) particles produced by Heck dispersion polymerization, (f) confocal microscopy image of PPV particles. () (The scale bars in (a)–(d) represent 2 µm and in (e) and (f) they represent 10 µm).

Figure 2.10 CPNs produced via Sonogashira coupling: (a) evolution of particle size with increasing monomer loading, (b) scanning electron microscope image of CPNs generated by Sonogashira dispersion polymerization (the scale bar represents 10 µm), (c) functionalization of CPNs with PEG and peptide (cRGD) by thiol‐yne click chemistry. ()

Chapter 3: Conjugated Polymer Nanoparticles and Semiconducting Polymer Dots for Molecular Sensing and

In Vivo

and Cellular Imaging

Scheme 3.1 The preparation of CPNs with three different methods, including nanoprecipitation, miniemulsion, and self‐assembly method.

Figure 3.1 Direct functionalization of Pdots with different side‐chain carboxyl groups. Bioconjugation was performed on the carboxyl groups. The Pdot‐bioconjugates were specific for cellular labeling.

Figure 3.2 (a) Surface functionalization of PFBT Pdots with the amphiphilic polymer PS–PEG‐COOH. (b) Surface functionalization of CNPPV Pdots with the amphiphilic polymer PSMA. (c) Encapsulation of Pdots with polyelectrolytes. (i–ii) Schematic illustration of Pdots encapsulation by one or two polyelectrolytes. (iii) Bioconjugation of streptavidin onto the surface of the polyelectrolytes encapsulated Pdots for cell labeling and imaging.

Figure 3.3 Rhodamine spirolactam‐doped CPNs for mercury ion detection.

Figure 3.4 (a) Schematic illustration of the conjugated polymer dots for oxygen sensing. (b) Schematic illustration of the formation of oxygen‐sensitive Pdots with glucose‐oxidase modification for the monitoring of glucose levels

in vivo

.

Figure 3.5 (a) Schematic of the preparation of CL CPNs for H

2

O

2

detection by doping the chemiluminescent substrate into the CPNs through hydrophobic interactions. (b) Schematic illustration of CL CPNs for detection of superoxide anion by grafting conjugated polymer with imidazopyriazinone.

Figure 3.6 Schematic illustrating three routes to prepare pH‐sensitive Pdots.

Figure 3.7 Preparation of RhB–Pdots for temperature sensing.

Figure 3.8 Protease detection using CPNs. (a) Sensing scheme for protease. The cross‐linked CPNs showed quenched state. The cleavage of the linker by protease released the strain and turned on the fluorescence. (b) Fluoresce spectra of the cross‐linked CPNs before (red) and after (blue) incubation with trypsin.

Figure 3.9 (a) Schematic showing the sensing of fluoride anion by coprecipitation of PPE and coumarin. (b) Fluorescent tubes of the selective detection of fluoride anion over other anions under a 365 nm lamp.

Figure 3.10 BODIPY‐based multicolor Pdots with narrow emission for fluorescence imaging of the EpCAM receptors on the surface of MCF‐7 cells.

Figure 3.11 Preparation of squaraine‐based Pdots with narrow NIR fluorescence emission for specific cellular labeling.

Figure 3.12 (a) Schematic illustrating the formation of blended NIR‐emissive Pdots conjugated with CTX for brain tumor imaging. (b) Fluorescence imaging of the healthy wild type mice (left) and medulloblastoma tumors in ND2:SmoA mice (right) after tail‐vein injection of the Pdots.

Figure 3.13 Schematic of self‐luminescing BRET‐FRET NIR‐emissive CPNs for

in vivo

imaging.

Figure 3.14 (a) Schematic illustration of the preparation of self‐quenched CPNs through nanoprecipitation. (b) Bioconjugation of the RGD on the surface of the self‐quenched CPNs and photoacoustic images of tumors after systemic injection of the self‐quenched CPNs‐RGD (SPN10‐RGD) and the bare CPNs (SPN10).

Figure 3.15 Schematic illustration of multifunctional theranostic CPNs for liver cancer diagnosis and therapy.

Chapter 4: Conjugated Polymers for

In Vivo

Fluorescence Imaging

Figure 4.1 (a) Chemical structure of PFBTDBT10. (b) Schematic illustration of CP‐FA dot. (c) Normalized UV–vis absorption (solid) and emission spectra (dashed) of CP dots in water (excited at 465 nm). (d)

In vivo

fluorescence imaging of H22 tumor‐bearing mice at different time points after intravenous injection of CP‐FA dots and CP dots, respectively. The tumor‐bearing mouse model was established by subcutaneous inoculating murine hepatic H22 cancer cells into the mouse left axillary space. The white circles indicated the tumor mass. (e)

Ex vivo

fluorescence imaging of various mouse tissues at 24 h post intravenous injection of CP‐FA dots and CP dots, respectively. ()

Figure 4.2 (a) Chemical structures of PSMA, PFBT, and PF‐DBT5 as well as schematic illustration of formation of functionalized CP dots (CP dots‐PEG/CTX). (b) UV–vis absorption and emission spectra of the CP dots. (c)

Ex vivo

fluorescence imaging of healthy brains in wild type mice and medulloblastoma tumors in ND2:SmoA1 mice at 72 h post intravenous administration of CP dots‐PEG/CTX or CP dots surface functionalized with only PEG (CP dots‐PEG). The white circle indicates the tumor site. (d) Tumor targeting efficiency by quantification of the fluorescence intensities in ND2:SmoA1 versus wild type mice and cerebellum versus frontal lobe. ()

Figure 4.3 (a) Chemical structures of PCFDP and IR755COOH as well as schematic illustration of the PCFDP NPs and NanoDRONE formation and the ROS sensing. (b) UV–vis absorption and (c) emission spectra of PCFDP NPs and NanoDRONE in PBS buffer. (d) Fluorescence changes of PCFDP NPs, QD655, and Cy5.5 at 678, 655, and 693 nm, respectively, when addition of different concentrations of ClO

‐

in PBS. [PCFDP NPs] = [Cy5.5] = 1.0 µg; [QD655] = 20 μM. Excitation: 405 nm for PCFDP NPs; 500 nm for QD655; 630 nm for Cy5.5. (e) PL spectra of NanoDRONE in PBS buffer with and without addition of ONOO

‐

at concentrations of 0.1, 0.2, 0.3, 0.4, and 0.5 μM, respectively. (f) Fluorescence changes of NanoDRONE (0.1 µg ml

−1

) in the presence of various ROS in nitrogen‐purged PBS.

F

and

F

0

stood for the PL intensities at 678 nm with and without addition of 1 μM of ROS. Excitation at 405 nm. (g)

In vivo

fluorescence imaging of ROS using NanoDRONE in LPS‐induced peritonitis‐bearing mice. LPS was intraperitoneally administrated into the mice to establish the peritonitis‐bearing mouse model. As a control, saline was also intraperitoneally injected into another group of mice. (h) Quantitative analysis of the fluorescence signals in the peritoneal cavity based on the imaging in (g). * represented

P

 < 0.05 between the two groups. ()

Figure 4.4 (a) Chemical structure of PFBD and schematic illustration of Tat‐PFBD dot. (b) UV–vis absorption and emission spectra of Tat‐PFBD dots in water. Excitation at 474 nm. (c) Confocal image of MSCs after incubation with Tat‐PFBD dots (4 nM) at 37 °C for 4 h. (d) Time‐dependent

in vivo

fluorescence imaging of skin wound‐bearing mice. The left side wound was transplanted with Tat‐PFBD dot‐labeled MSCs; the right one was transplanted with nonlabeled MSCs. (e) Quantitative analysis of the fluorescence intensity change at the left wound site. (f) Corresponding

in vivo

bioluminescence imaging of the same mouse in (a). (g) Quantitative analyses of the bioluminescence intensity changes at left and right wound sites. (h–j) Confocal images of the regenerated skin from mice treated with Tat‐PFBD dot‐labeled MSCs for 21 days. GFP was intrinsically expressed in MSCs for (h); CD31 was costained to visualize blood vessels for (i); VEGF and bFGF was costained, respectively, to reveal paracrine cytokines of Tat‐PFBD dot‐labeled MSCs for (j). The white arrows indicate the Tat‐PFBD dots in (h–j). 4′,6‐Diamidino‐2‐phenylindole (DAPI) was used to stain cell nuclei in (c, h–j). ()

Figure 4.5 (a) Chemical structure of F127 and schematic illustration of the PFBT‐F127‐SiO

2

NP formation. (b) TEM image of PFBT‐F127‐SiO

2

NPs. (c) TPA spectra of PFBT‐F127‐SiO

2

NPs on the basis of CP chain concentration and Evans Blue in water. (d) Intravital TPFI of mouse brain vasculature at different vertical depths after retro‐orbital injection of PFBT‐F127‐SiO

2

. ()

Figure 4.6 (a) Chemical structure of PFVBT and schematic illustration of the FMCPNP. (b) UV–vis absorption and emission spectra of FMCPNPs in water. Excitation at 518 nm. (c) Hysteresis curve of FMCPNPs. (d) Time‐dependent

in vivo

noninvasive fluorescence imaging of mice after intravenous injection of FMCPNPs. (e)

In vivo

MRI of the same mouse in (d) preinjection and postinjection of FMCPNPs for 5 h. The arrow in (d) and the circle in (e) indicates the tumor site. ()

Figure 4.7 (a) Schematic illustration of the formation of various cvPDs. (b) Mass extinction coefficient and PL spectra of NIR‐cvPDs in water. (c) Time‐dependent

in vivo

fluorescence imaging of mice after intradermal injection of NIR‐cvPDs (10 µl of 1.7 mg ml

−1

) into the right paw. The arrows and arrowheads indicate axillary lymph node and lymphatic vessels, respectively. ()

Chapter 5: π‐Conjugated/Semiconducting Polymer Nanoparticles for Photoacoustic Imaging

Figure 5.1 Schematic illustration of mechanism of PA imaging.

Scheme 5.1 (a) Chemical structures of some SPs and small molecules used for PA imaging. (b) Chemical structures of some amphiphilic molecules used for the preparation of SPNs.

Figure 5.2 (a) Schematic illustration of preparation of SPN1. (b) Absorption spectrum of SPN1 (50 µg ml

−1

) in water. (c) PA amplitude of SPN1 (50 µg ml

−1

) in agar gel embedded in chicken breast muscle as a function of depth from the laser irradiated surface. (d) PA imaging of brain after systemic administration of SPN1 for 0, 5, and 60 min. (.)

Figure 5.3 (a) Schematic illustration of the preparation of SPN2 using DSPE‐mPEG

2000

as the matrix. (b) UV–vis absorption of SPN2 and GNR before and after laser irradiation for 5 s and 6 min, respectively. (c) The PA intensity as a function of concentration of SPN2. The inset figure shows the PA image of SPN2 at the concentration of 0.5 mg ml

−1

. (d) PA images of rat cortical vasculature before and after systemic administration of SPN2 for 10 min. (.)

Figure 5.4 (a) Schematic illustration of preparation of SPNs via nanoprecipitation. (b) PA spectra of SPNs in 1 × PBS (pH = 7.4). (c) PA intensities of SPNs as a function of concentration in PBS (pH = 7.4). (d) Normalized PA and fluorescence intensities based on same mass extinction coefficients of SPNs at 710 nm. (e) PA and PA/ultrasound coregistered images of SPN6‐matrigel inclusions in mice at different concentrations. (f) PA signals at 750 nm in tumor as a function of time after administration of SPN6. The error bars represent standard deviations of three separate measurements. (.)

Figure 5.5 (a) Schematic illustration of preparation of SPN3/Ce6@lipid‐Gd‐DOTA micelles. (b) Absorption spectra of Ce6 (in DMSO), SPN3@lipid micelles, and SPN3/Ce6@lipid‐Gd‐DOTA micelles. (c) PA images and (d) PA intensities of SPN3/Ce6@lipid‐Gd‐DOTA micelles under different concentrations. (e) PA images of the tumor sites before and after intratumoral injection of SPN3/Ce6@lipid‐Gd‐DOTA micelles. (.)

Figure 5.6 (a) Schematic illustration of preparation of SPNs through nanoprecipitation method. (b) PA amplitudes of the matrigel‐containing solutions of nanoparticles in the subcutaneous dorsal area of living mice as a function of mass concentration. The background signal was 0.21 ± 0.03 a.u., which were calculated as the average PA signal in the areas where no nanoparticles were injected.

R

2

 = 0.992, 0.990, and 0.992 for SPN3, SWNTs, and GNR, respectively. (c) Ultrasound (upper) and PA/ultrasound coregistered (lower) images of lymph nodes after intravenous injection of SPN3 (50 µg mouse

−1

). Images represent the transverse slices of the lymph nodes. The white arrows and dashed circles showed the lymph nodes. The abbreviation names BLN, ILN, and SCLN represent brachial lymph node, inguinal lymph node, and superficial cervical lymph node, respectively. (d)

Ex vivo

PA/ultrasound coregistered (top) and fluorescence/bright‐field (bottom) images of resected lymph nodes from SPN3‐injected mouse (left) and saline‐injected mouse (right) in an agar phantom. (.)

Figure 5.7 (a) Schematic illustration of RSPN probe for PA imaging of ROS. (b) Representative PA spectra of RSPN (5 µg ml

−1

) in the absence and presence of different ROS (5 μM). (c) Quantification of PA ratio (PA

700

/PA

820

) for RAW264.7 cell pellets with and without LPS/IFN‐γ or LPS/IFN‐γ/NAC treatment. Error bars represents standard deviations from four measurements.

*

Statistically significant difference in PA

700

/PA

820

between LPS/IFN‐γ treated and untreated or LPS/IFN‐γ/NAC‐treated cell pellets. (

*

p

 < 0.05) (d) PA and ultrasound overlaid images of saline‐ (i) and zymosan‐treated (ii) regions in the thigh of mice (

n

 = 3). RSPN (3 µg in 50 µl) was intramuscularly injected into the thigh of mice after treatment of zymosan for 20 min. (.)

Figure 5.8 (a) Schematic illustration of fabrication process of IONP@SPN1‐PEG nanocomposites. (b) Absorption spectra of IONP@SPN1 and IONP@SPN1‐PEG at same concentration of SP1 (0.01 mg ml

−1

). (c) PA images of mice before (left) and after systemic administration of IONP@SPN1‐PEG for 24 h (right). (d) Quantification of PA signals from the tumor area in figure (c). (.)

Chapter 6: Conjugated Polymers for Two‐Photon Live Cell Imaging

Figure 6.1 (a) Jablonski diagram illustrating 1PA, 2PA, and the related fluorescence process. Deep tissue penetration length of NIR light. (b) The NIR window for

in vivo

imaging. The spectral response of oxygenated hemoglobin, deoxygenated hemoglobin, and water varies as a function of wavelength. The red highlighted area indicates the biological optical window where adsorption due to the body is minimum. ()

Figure 6.2 (a) Chemical structure of PFBD and DSPE–PEG, and the schematic illustration of CPN formation at different feed concentration. (b) Confocal images of MCF‐7 breast cancer cells at different designated generations after overnight incubation with 2 nM of CPN‐C‐TAT, CPN‐N‐TAT, CPN‐M‐TAT, or Qtracker

®

585 at 37 °C. ()

Scheme 6.1 Highly fluorescent two‐photon conjugated polymers. ()

Figure 6.3 Chemical structures of PPE 1, PPE 2, PTAA, POWT, and POMT. (a) 2PEM of PPE derivatives interacting with live mouse fibroblast cells. Cells were incubated with PPE 1 (left,

λ

ex

 = 710 nm) or PPE 2 (right,

λ

ex

 = 760 nm) in DMEM at 37 °C for 4 h. (b) Micrographs of insulin amyloid aggregates stained with POMT. 2PEM of reduced (left) and nonreduced insulin amyloid at 820 nm. ()

Scheme 6.2 Molecular structures and

δ

values per RU at 800 nm of a few polyfluorene‐based cationic conjugated polymers.

Figure 6.4 (a) Wavelength‐dependent

δ

Φ

F

(per RU) values of PFV, PFVMO, and PFVCN in water. The inset shows their photographs under fs laser excitation at 810 nm. (b) Normalized absorption (solid lines) and fluorescence spectra (dash lines) of PFV, PFVMO, and PFVCN in water. Overlaid images of 2PEF and bright‐field images of HeLa cancer cells treated with PFV (c), PFVMO (d), and PFVCN (e) for 3 h. Scale bar: 10 µm. (f) Metabolic viability of HeLa cells after incubation with PFV, PFVMO, PFVCN, and TMPyP4 of different concentrations (in RU for polymers) for 24 h. ()

Figure 6.5 Chemical structures of PDHF, MEH‐PPV, PFPV, and PPE. (a) Photograph of fluorescence from aqueous CPNs dispersions under 2PE by a 800 nm mode‐locked Ti:sapphire laser. (b) Fluorescence images (days 1, 2, and 3) of cultured cells in a microfluidic device. 2PE images showing projection of capillary structures through 80 mm of the central gel region. ()

Figure 6.6 Chemical structures of poly‐1 to poly‐5. Confocal fluorescence micrographs of fixed HeLa cells labeled with poly‐1 (green, inside) and poly‐4 (red, outside) NPs excited at 780 nm. ()

Figure 6.7 (a) Illustrative preparation procedures of CPNs; (b) chemical structures of DPSB, encapsulation matrix PSMA, and polymer S1, S2, M1, M2; (c) 2PE fluorescence spectra of CPNs with the same concentration (2.5 μM RU) in water dispersion,

λ

EX

 = 810 nm; (d) wavelength‐dependent

δ

and

δ

Φ

F

values per RU for different CPNs; (e–j) Bright‐field (e, h), 1PE fluorescence (f, i) and 2PE fluorescence (g. j) images of HepG2 cells after incubation with PMSA‐M2 CPNs (top line) and CellMask Deep Red Plasma membrane stain (bottom line).

λ

EX

is 405 (f), 650 (i), and 750 nm (g and j), respectively. ()

Figure 6.8 (a) Structure of the O

2

‐sensitive conjugated polymer, (b) their conformation in organic solvent and in nanoparticle form after precipitation with water, TEM image of the resulting NPs, and ratio‐metric intensity imaging of NPs in MEF cells exposed to different O

2

at 37 °C and treated with inhibitor of respiration. ()

Figure 6.9 (a) Schematic preparation procedures of MgPc/PFV NPs; 1PE (b) and 2PE (c) fluorescence spectra of PFV NPs and MgPc/PFV NPs. Excitation wavelengths for 1PE and 2PE are 437 and 800 nm, respectively; (d) cell viability assay of HepG2 cancer cells treated with MgPc(0.75%)/PFV NPs and MgPc NPs containing the same amount of MgPc for 8 h; (e) overlaid 2PE fluorescent images and bright‐field images of HepG2 cancer cells after incubation with 2 μM MgPc(0.75%)/PFV NPs. The nanoparticle concentration is in terms of the PFV per RU. ()

Figure 6.10 (a) Chemical structures of the CP with 2PEF feature (P‐F8‐DPSB), NIR fluorescent dye (DPA‐PR‐PDI) used for the construction of the hybrid CPNs and 2PEF imaging of HeLa cells; (b) normalized absorption and fluorescence emission spectra of P‐F8‐DPSB polymer and DPA‐PR‐PDI in THF solution; (c) fluorescence emission spectra of the hybrid P‐F8‐DPSB/DPA‐PR‐PDI NPs with various polymer/dye ratios upon 800 nm excitation. ()

Figure 6.11 (a) Chemical structure of F127 and PFBT; (b) schematic illustration of the fabrication of PFBT‐F127‐SiO

2

NPs. (c) 2PA spectra of PFBT‐F127‐SiO

2

NPs (based on CP chain concentration) and Evans blue in water. (d) UV–vis (dash‐dotted line) and PL (solid line) spectra of PFBT‐F127‐SiO

2

NPs (red) and PFBT‐DSPE NPs (black) at 25 μg ml

−1

of PFBT. Fluorescence decay curves of PFBT‐F127‐SiO

2

NPs (red) and PFBT‐DSPE NPs (black). (e) Intravital TPFI of PFBT‐F127‐SiO

2

NPs stained blood vessels of mice brain at various depths (A–G) and the respective Z‐projected image (H) as well as 3D image (I). Scale bar: 50 µm. ()

Chapter 7: Water‐Soluble Conjugated Polymers for Sensing and Imaging Applications

Figure 7.1 (a) Scheme of the DNA detection based on FRET between

CP 1

and PNA‐C*. () (b) Scheme of the detection of DNA methylation based on FRET between dGTP‐Fl and

CP 2

. ()

Figure 7.2 (a) Scheme of label‐free detection of DNA‐binding protein based on the FRET between PFEP and Sybr Green I. () (b) Scheme of the label‐free detection of thrombin based on G‐quadruplex specific dye (ThT). ()

Figure 7.3 Schematic illustration of FRET‐based cell sensing using CP‐GFP complexes. ()

Figure 7.4 Schematic drawings of (a) “light‐on,” (b) “light‐off,” and (c) “two‐way” hybrid sensors for the detection of protein‐DNA interactions by fluorescence recovery, fluorescence quenching, or both ways. ()

Figure 7.5 (A) Schematic presentation of DNA analysis with the GO–CPs hybrid system. () (B) Scheme of a GO and CP 4 hybrid probe for the detection of conformation changes of calmodulin (CaM).

Figure 7.6 (A) Schematic description of the formation of polythiophene/single‐stranded nucleic acid duplex and polythiophene/hybridized nucleic acid triplex forms. (B) Photographs of solutions of (a) polythiophene, (b) polythiophene/X1 duplex, (c) polythiophene/X1/Y1 triplex, (d) polythiophene/X1/Y2 mixture, and (e) polythiophene/X1/Y3 mixture.

Figure 7.7 (a) Schematic illustration of telomerase activity detection using the hydrophilic/hydrophobic properties of the bipolar probes. (b) Schematic illustration of the strategy of Hg

2+

detection by tuning the aggregate state of the bipolar probe. ()

Figure 7.8 (a) First row: single‐plane confocal micrograph of

E. coli

cells stained with ZCOE and DSSN+. Scale bars are 5 µm. Second and third row: two single‐plane confocal and bright‐field (BF) micrographs of yeast cells stained with ZCOE, propidium iodide, and DSSN+. Arrow indicates axial attenuated emission from DSSN+ when illuminated by the polarized light. Four scale bars are 10 µm. (b) Single‐plane confocal micrographs of COS‐1 cells stained with ZCOE (Top). Dual stain with DSSN+ imaged soon after staining and after ∼12 h (middle). Dual stain with LysoTracker Green applied 12 h after ZCOE (bottom). Scale bars are 10 µm. ()

Figure 7.9 Molecular structure, UV–vis absorption and fluorescence emission spectra of IR‐E1 (upper left two figures). Fluorescence imaging of a typical Hipco CNT and IR‐E1 (upper right figure). NIR‐II dynamic imaging of hypoperfusion in a TBI mouse at single‐vessel resolution. (a–f) Video rate dynamic image frames recorded in the injured region of a TBI mouse (2 h after TBI) in the hypoperfusion phase with 300 ms exposure time in the NIR‐II window (>1300 nm) after intravenous injection of IR‐E1 (10 mg kg

−1

), showing ultraslow blood perfusion from the transverse sinus region (bright lower‐left region) into the injured region. Blue arrow points to the perfusion front over which the signal traveled. (f–h) A single vessel was observed to be perfused over ≈2.9 min. Red arrows point to the blood flow in front of the vessel.

Figure 7.10 Chemical structures of F127 and PFBT, schematic illustration of the fabrication of PFBT‐F127‐SiO

2

NPs, and intravital TPFI of PFBT‐F127‐SiO

2

NPs stained blood vessels of mice brain at depth of 0 µm (a), 50 µm (b), 100 µm (c), 200 µm (d), 300 µm (e), 400 µm (f), and 500 µm (g), and the respective Z‐projected image (h) as well as 3D image (i). All the images share the same scale bar of 50 µm. ()

Figure 7.11 (a) Fluorescence spectra of various conjugated polymer dots. (b) Differential interference contrast (DIC) images (top), and fluorescence images (bottom) of macrophage cells labeled with PPE, PFPV, PFBT, and MEHPPV dots, respectively. The scale bar is 10 µm. ()

Figure 7.12 Chemical structures and the emission colors of

P1

,

P2

,

P3

, and

P4

. (a) Fluorescence microscope images of

E. coli

‐CPN microparticles. The excitation wavelength is 380/30 nm for

P1

, 455/70 nm for

P2

, and 540/40 nm for

P4

. The scale bar is 20 µm. (b) SEM images of

E. coli

and

E. coli

‐CPN microparticles. The scale bar is 1.0 µm. (c) Various color‐barcoded microparticles by mixing the

E. coli

and the CPNs. The image was taken under 365 nm UV light.

Figure 7.13 (A) Chemical structures, normalized UV–vis absorption (dashed line) and PL spectra (solid line), and photographs under a hand‐held UV lamp (excited at 365 nm) of SiO

2

@CP@SiO

2

NPs. (B) Confocal fluorescence images of SKBR‐3 breast cancer cells (a and b) and NIH/3T3 fibroblast cells (c and d) after 2 h incubation with SiO

2

@PFBT@SiO

2

‐Pep (a and c) and SiO

2

@PFBT@SiO

2

‐COOH (b and d) NP suspensions at 100 mg ml

−1

NPs at 37 °C. All images have the same scale bar of 40 µm. ()

Figure 7.14 (a) The preparation of multicolor CPNs (P1–4/PSMA) and their modification with an antibody. (b) Multichannel fluorescence images of MCF‐7 cells with P1–4/PSMA/anti‐EpCAM CPNs. The excitation wavelengths are 405 nm, 488 nm, and 559 nm.

Figure 7.15 (a) Confocal images of the MCF‐7 breast cancer cells after 4 h incubation with the MCPNP (left), FMCPNP (middle), and free folic acid‐pretreated MCF‐7 breast cancer cells after 4 h incubation with 0.25 mg ml

−1

of FMCPNPs (right). The nuclei were stained by DAPI. (b)

In vivo

fluorescence images of mouse injected with FMCPNPs acquired at 1, 6, and 18 h post injection (upper). Fluorescence images of various organs from the mice treated with FMCPNPs (left) and MCPNPs (right), respectively, at 12 h post injection. (c)

T

2

‐weighted MR images of FMCPNPs in water. (d)

T

2

relaxivity plot of aqueous suspension of FMCPNPs. (e) Representative

in vivo

MR images of mouse pre injection (left) and post injection with FMCPNPs for 5 h (right).

Figure 7.16 (a) Chemical structure of HCPE. (b)

In vivo

noninvasive fluorescence imaging of H

22

tumor‐bearing mice after intravenous injection of HCPE‐Gd. (c)

T

1

‐weighted MR images of HCPE–Gd and Magnevist at various concentrations of Gd(III). (d) Water proton longitudinal relaxation rate (1/

T

1

) of HCPE–Gd and Magnevist as a function of Gd(III) concentration. (e)

T

1

‐weighted MR images of H

22

tumor‐bearing mice after intravenous injection of HCPE‐Gd at 0 and 6 h post injection. The red circles indicate the tumor site.

Figure 7.17 (a) CLSM fluorescence and bright‐field images of fixed MCF‐7 breast cancer cells after 2.5 h incubation with FPBT NC (A, B), FPAuBT NC2 (C, D), PBT NC (E, F), and PAuBT NC (G, H). (b) FE‐TEM images of FPAuBT NCs prepared by encapsulation of 1 mg PFVBT with 1 mg (A), 2 mg (B), and 4 mg (C) of Au NPs, respectively, in PLGA matrix (upper). Dark‐field microscopic images of MCF‐7 breast cancer cells after 2.5 h incubation with FPAuBT NC2s (A) and PAuBT NC2s (B) and control cells without nanocomposites (C) (bottom).

Figure 7.18 (A) Color dark‐field (a) and TEM images of silver‐coated GNRs (b). TEM imaging of silica‐coated SGNR (c). The dark‐field (d) and fluorescence (e) images of PFBT‐coated SSGNRs. The merged image from the dark‐field (red) and fluorescence (green) channels (f). (B) Single particle fluorescence imaging of regular PFBT nanoparticles (a) and PFBT‐SSGNRs (b) represent intensity tracks as a function of time from individual nanoparticles. (C) Representative single particle dark‐field images of PFBT‐SSGNRs on cell membrane as a function of time. ()

Chapter 8: Conjugated Polymers for Gene Delivery

Figure 8.1 Obstacles hampering the effective delivery of nucleic acids by nonviral carriers. Various nonviral agents may be used to deliver different genetic materials, such as DNA, mRNA, and small interfering RNA (siRNA) or microRNA (miRNA). For successful gene delivery, the nonviral vectors need to protect these genetic materials from enzymatic degradation in cellular environments and rapid renal clearance from the circulation system. These nonviral gene carriers must also facilitate the cellular endocytic internalization and endosomal escape of the nucleic acids. Both siRNA and miRNA have to be encapsulated by the RNA‐induced silencing complex (RISC) while mRNA must associate with the translational machinery. DNA, in contrast, has to be transported into the nucleus to induce its function. (.)

Figure 8.2 Specific intracellular targeting of conjugated polymer nanoparticles toward mitochondria. (a) Chemical structures of PPEs in the presence and absence of biodegradable linkers on the polymeric backbones (i.e., PPE‐1 and PPE‐2, respectively) accompanied by a schematic illustration demonstrating the cellular entry of biodegradable CPN (i.e., CPN‐1), followed by its mitochondrial localization. (b) Fluorescence microscopic images showing the CPN/HA‐incubated HeLa cells (green). The mitochondria and nuclei of the HeLa cells were stained in red and blue, respectively. Scale bars represent 20 µm. (c) Signal intensity of the fluorescently‐stained mitochondria and cytosol of HeLa cells treated with CPNs. The higher fluorescent intensity of mitochondria of HeLa cells incubated with CPN‐1 shows the specific localization of CPN‐1 toward mitochondria. (.)

Figure 8.3 Development of polycation‐based nanoparticles with low molecular weight for the effective co‐delivery of drug and pDNA. (a) Schematic illustration showing the synthesis of BIP‐PGEA‐CPT/pDNA complexes. (b–g) HepG2 cells incubated with: (b) free antitumor drug CPT with a concentration of 4.5 µg ml

−1

equal that of CPT in BIP‐PGEA2/CPT nanoparticles at an N/P ratio of 25, (c) free prodrug 5‐FC, with a concentration of 80 µg ml

−1

, (d) BIP‐PGEA2 nanoparticles, (e) BIP‐PGEA2‐CPT nanoparticles, (f) BIP‐PGEA2/CD/5‐FC nanoparticles, with a 5‐FC concentration of 80 µg ml

−1

and at an N/P ratio of 25, and (g) BIP‐PGEA2‐CPT/CD/5‐FC nanoparticles, with a 5‐FC concentration of 80 µg ml

−1

and at an N/P ratio of 25. The results demonstrated the strong synergistic antitumor effect induced by BIP‐PGEA‐CPT nanoparticles on HepG2 cells. (.)

Figure 8.4 Brush‐like cation conjugated polyelectrolyte nanoparticles for the effective visualized delivery of siRNA. (a) Schematic illustration showing the delivery of siRNA using PFNBr. (b–d) Confocal microscopy images of PANC‐1 cells incubated with individual Cy3‐labeled siRNAs, CPNs, and their polyplexes for 4 h, illustrating their cellular internalization: (b) Cy3‐labeled siRNA (100 nmol l

−1

); (c) PFNBr (40 nmol l

−1

); (d) Cy3‐labeled siRNA (100 nmol l

−1

) and PFNBr (40 nmol l

−1

). PFNBr and cy3‐labeled siRNA were represented in blue and red, respectively. (.)

Figure 8.5 Multifunctional poly(

p

‐phenylene vinylene)‐based structure for the light‐enhanced delivery of siRNA. (a) Schematic illustration demonstrating the high siRNA delivery based on the white light‐facilitated endosomal escape approach. (b) Endosomal escape of PPV/siRNA polyplexes in the absence and presence of white light irradiation. (c) Colocalization of the PPV/siRNA polyplexes with endosome after another 20 h incubation post white light irradiation. Lysotracker, siRNA, and PPV were represented in blue, green, and red, respectively. (d) Transfection efficiency of PPV/siRNA polyplexes with respect to those of positive controls, that is, PEI and lipofectamine 2000. (e) Luciferase intensity measured from HeLa‐Luc cells after a 24 h incubation with PPV/siRNA polyplexes at 1 µg anti‐Luc siRNA. (.)

Chapter 9: Conductive Polymer‐Based Functional Structures for Neural Therapeutic Applications

Figure 9.1 Conductive polymer‐based functional structures, such as hydrogels and nanofibers, for various applications in neural therapeutic applications, specifically electrostimulated drug delivery, neural cell and tissue scaffolds, and implantable neural prostheses.

Figure 9.2 Conjugated backbone structure of conducting polymer comprising alternating single and double bonds, in which π bond facilitates the delocalization of electrons while σ bond maintains the strength of the chain.

Figure 9.3 Common conductive polymers with their chemical structures, that is, polypyrrole (PPy), polyaniline (PAni), polythiophene (PTh), and poly(3,4‐ethylenedioxythiophene) (PEDOT), used for biomedical applications, specifically for neural therapy and regenerative medicine.

Figure 9.4 General strategies for the synthesis of electroconductive polymer‐based hydrogels based on electrochemical and chemical oxidative polymerization approaches.

Chapter 10: Conjugated Polymers for Photodynamic Therapy

Figure 10.1 A timeline of selected milestones in the historical development of PDT.

Figure 10.2 Schematic representation of PDT process.

Figure 10.3 Two types of oxygen sensitization mechanism provided by CPs.

Figure 10.4 Jablonski diagram showing the formation of singlet oxygen.

Figure 10.5 Light penetration through tissue. The penetration depth of typical light is dominated by the rates of absorption, scattering, transmission, and reflection by the tissues, which vary with different wavelengths.

Scheme 10.1 Far‐red‐ and near‐IR‐emitting conjugated polymers used in PDT.

Scheme 10.2 Conjugated polymers as energy transfer systems to Porphyrin dyes for PDT.

Scheme 10.3 Hybrid photosensitizers based on conjugated polymers for PDT.

Figure 10.6 (a) Structure of cationic polymer PPE and (b) growth curves of

E. Coli

bacteria with and without the presence of CP15 for 16 h under light irradiation at 560 nm.

Figure 10.7 Structure multiple QA groups functionalized CPEs.

Figure 10.8 Mechanism of biocidal activity of PPE grafted on silica colloidal particles. (i) The first step is either reversible physisorption or covalent grafting of CP onto the surface of silica colloidal particles to form polymer beads and they entrap bacteria inside the beads under dark conditions. In second step (ii) illumination of light leads to production of ROS from polymer beads to kill of bacteria in step (iii) and to degrade bacterial membranes (iv). Finally, (v) large aggregated assemblies of dead bacterial cells associated with the polymer beads collected as debris.

Figure 10.9 CLSM image of a microcapsule cluster 10 min after introduction into a solution of

P. aeruginosa

kept in the dark. (b) Interior image of microcapsule cluster, showing bacteria entrapped within the cluster and killed after 1 h of exposure to white light. (Red – alive bacteria; green – dead bacteria).

Figure 10.10 Chemical structure of CP20 polymer.

Figure 10.11 Structure and antimicrobial activity of PPV polymer.

Figure 10.12 CLSM images of Jurkat T cells and Gram‐positive

B. Subtilis

incubated with CP21 (a) and Jurkat T cells and Gram‐negative ampicillin‐resistant

E. coli

incubated with CP21 (b). Left: bright field images, right: fluorescent images.

Figure 10.13 (a) Binding and biocidal activity of PT/TPPN complex and (b) % killing efficiency CP8 compared with PTP and TPPN controls under dark and light irradiation.

Figure 10.14 Supramolecular antibiotic reversible switch.

Scheme 10.4 Chemical structure of PMNT.

Figure 10.15 Schematic representation of cancer cell death induced by ROS generation in PDT.

Figure 10.16 (a) Chemical structures of phosphorescent CPs with the Ir(III) complexes; (b) HR‐TEM of WPF‐Ir4 in aqueous solution; and (c) mechanisms illustrating oxygen sensing and PDT.

Figure 10.17 Schematic representation of mechanism of light manipulated drug‐uptake approach to combat drug resistance in cancer cells.

Figure 10.18 Lumol‐induced BRET transfer to PPV for effective PDT.

Figure 10.19 Schematic illustration of the combined functions of PDT and PTT in single system.

Figure 10.20 Illustration of thermoresponsive HCP@HPE unimolecular micelles and illustration of the combination of 2P‐FRET and photothermal effect of NIR for photodynamic therapy.

Chapter 11: Conjugated Polymers for Near‐Infrared Photothermal Therapy of Cancer

Figure 11.1 Polyaniline nanoparticles (PANP) serve as NIR‐absorbing photothermal agents within a tumor. (a) The scheme of the preparation of PANP. (b) Photographs of EB‐PANPs in culture medium, A431 cells in culture medium, A431 cells incubated with EB‐PANPs (transition to ES‐PANPs), and the free medium. (c) Absorption spectra of A431 cells treated with EB‐PANPs (transition to ES‐PANPs) and free EB‐PANPs in culture medium. ()

Figure 11.2 PEDOT:PSS–PEG nanoparticles showed ideal therapeutic efficacy under NIR laser irradiation. The preparation scheme (a) and biodistribution (b) of the stealth PEDOT:PSS nanoparticles. (c) Infrared thermal images of 4 T1 tumor‐bearing mice without (upper row) or with (lower row) intravenous injection of PEDOT:PSS–PEG (10 mg kg

−1

, 48 h pi) under 808 nm laser irradiation taken at different time intervals. The laser power density was 0.5 W cm

−2

. Arrows point to the tumors. (d) Growth of 4 T1 tumors in different groups of mice after treatment and survival curves of mice after various treatments. PEDOT:PSS–PEG‐injected mice after PTT treatment showed 100% survival ratio over 45 days. The relative tumor volumes were normalized to their initial sizes. For the treatment group, 10 mice injected with PEDOT:PSS–PEG at 48 h pi were exposed to the 808 nm laser (0.5 W cm

−2

, 5 min). The other three groups of mice were used as controls: untreated (control,

n

 = 8); laser only without PEDOT:PSS–PEG injection (laser only,

n

 = 14); injected with PEDOT:PSS–PEG but without laser irradiation (PEDOT:PSS–PEG only,

n

 = 10). Error bars were based on standard deviations. ()

Figure 11.3 The structure (a) and preparation scheme (b) of PFTTQ conjugated polymeric nanoparticles. ()

Figure 11.4 The synthesis and characterizations of IONP@PPy–PEG nanocomposite (a) a schematic showing the fabrication process of IONP@PPy–PEG nanocomposite (b) TEM images of IONP@PPy nanocomposite before and after PEGylation. Insets are higher resolution images. (c) Magnetization loops of IONP@PPy and IONP@PPy–PEG. Inset is a photo of an IONP@PPy–PEG solution placed nearby a magnet. (d)

T

2

‐weighted MR images (up) and the T2 relaxation rates (

R

2

) plot (bottom) of IONP@PPy–PEG solutions at different iron concentrations. ()

Figure 11.5 The

in vivo

bimodal imaging and photothermal therapy using IONP@PPy–PEG nanoparticles. (a)

T

2

‐weighted MR images of 4T1 tumor‐bearing mice i.v. injected with IONP@PPy–PEG (0.8 mg ml

−1

, 0.2 ml) taken at different time post injection. White circles highlight the tumor site. (b) Photoacoustic images of mice before injection and at 24 h post injection with IONP@PPy–PEG (0.8 mg ml

−1

, 0.2 ml). (c) Growth of 4T1 tumors in different groups of mice after various treatments indicated. The relative tumor volumes were normalized to their initial sizes. For the treatment group, six mice injected with IONP@PPy–PEG at 24 h post injection were exposed to the 808‐nm laser (1.5 W cm

−2

, 5 min). The other three groups of mice with six mice per group were used as controls: untreated; laser only; injected with IONP@PPy–PEG but without laser irradiation. (d) Survival curves of mice after various treatments indicated. IONP@PPy–PEG injected mice after PTT treatment showed 100% survival ratio over 40 days. ()

Figure 11.6 Conjugated polymer microparticles for selective cancer cell image‐guided photothermal therapy. Chemical structures of PFBT and PFTTQ (a), UV–vis (solid line) spectra of PFBT (blue, 0.01 mg ml

−1

) and PFTTQ (red, 0.01 mg ml

−1

), and PL spectrum of PFBT (dashed blue line) in DCM upon excitation at 460 nm (b). (c) Schematic illustration of CP MPs formation. (d) CLSM images of PFBT NP treated MCF‐7 and NIH‐3 T3 cells, PFBT MP treated MCF‐7 and NIH‐3 T3 cells. Both the cells were incubated with these particles at PFBT concentration of 30 µg ml

−1

for 6 h. The PFBT signal is collected above 505 nm upon excitation at 488 nm. The nucleus signal from DAPI is collected between 430 and 470 nm upon excitation at 405 nm. ()

Figure 11.7 Encapsulated conjugated oligomer nanoparticles for real‐time photoacoustic sentinel lymph node imaging. (a) Schematic illustration for the preparation of N4 NPs. (b) The PA intensity of N4 NPs upon laser excitation at different wavelength from 600 to 900 nm. The N4 NP concentration is 0.5 mg ml

−1

. (c) Real‐time PA imaging of SLN before and after N4 NPs injection. Excitation wavelength is 800 nm. BV stands for blood vessels. Scale bars: 1 mm. ()

Figure 11.8 The scheme and characterizations of PPy@BSA–Ce6(Gd) nanoparticles and the bimodal imaging

in vivo

. (a) A scheme showing the fabrication of PPy@BSA–Ce6 nanoparticles. (b) Temperature elevation of water and PPy@BSA–Ce6 solutions of different concentrations over a period of 5 min under exposure of the NIR light (808 nm,0.8 W cm

−2

). Detection of singlet oxygen by the SOSG test for different samples including free Ce6, PPy@BSA–Ce6, PPy@BSA, and BSA under the 660 nm light exposure (5 mW cm

−2

) for different periods of time. PPy@BSA–Ce6 showed largely retained light‐triggered SO generation ability compared with free Ce6 at the same Ce6 concentration (1 × 10

−6

 M). (c)

In vivo

fluorescence images of 4 T1 tumor‐bearing mice taken at different time points post injection of PPy@BSA–Ce6. The autofluorescence of the mouse was removed by spectral unmixing. (d) A scheme showing the fabrication PPy@BSA–Ce6 (Gd). (e) T1‐weighted MR images of 4 T1 tumor‐bearing mice before injection and 6 h after i.v. injection with PPy@BSA–Ce6(Gd). White circles highlight the tumor site. ()

Figure 11.9 The scheme of Fe

3

O

4

@PPy–PEG–DOX multifunctional nanoplatform (a) and the

in vitro

(b) and

in vivo

evaluations on the effects of the external stimuli on the cellular uptake of DOX, MR imaging (c) and the synergistic therapeutic efficacy (d). ()

Figure 11.10 Schematic illustration of the two‐photon laser‐regulated combination photothermal and chemotherapy with fast drug release (a); chemical structure of PLL‐

g

‐PEG/DNQ and its hydrophilic counterpart after Wolff rearrangement in aqueous media upon two‐photon laser irradiation (b); size distribution and TEM image (inset) of PFTTQ/DOX NPs in aqueous media before and after NIR laser irradiation. ()

Chapter 12: Conjugated Polymers for Disease Diagnosis and Theranostics Medicine

Figure 12.1 Chemical structures of some reported CPs and oligomers.

Figure 12.2 (a) Structure of newly developed PPE derivatives P1 and P2. Confocal laser scanning microscopy (CLSM) images of (b) P1 and (c) P2 with mannose‐binding

E. coli

strain ORN 178. ()

Figure 12.3 Structure of (a) oligomer FBT and (b) its application in the detection of ConA using GO/FBT hybrid probe. ()

Figure 12.4 Structures of (a) two cationic polymer PFP and PPV used (b) for the detection of pathogens based on FRET. ()

Figure 12.5 (a) Structure of cationic polymer PPVE and (b) its adsorption [PPVE = 5 μM] on the surfaces of

E. coli

,

B. subtilis

, and

C. albicans

in different concentrations of PBS. The corresponding numbers on the images depict the emission intensity values as determined by DVC View and Microsoft Excel softwares. ()

Figure 12.6 Hallmarks of cancer‐distinctive and complementary capabilities that enable tumor growth and metastatic dissemination.

Figure 12.7 Schematic demonstration of CCP‐based ssDNA sequence detection with a specific PNA‐Fl optical reporter probe, and the corresponding emission spectra for complementary (a) and noncomplementary (b) DNA by the excitation of conjugated polymers. ()

Figure 12.8 The distribution of

E

for RASSF1A promoter of 46 samples measured by the CCP‐based FRET technique, and the corresponding values and sum of the values for RASSF1A, OPCML, and HOXA9 genes. ()

Figure 12.9 Principle of LCR and lambda exonuclease‐assisted CCP biosensing for miRNA detection. ()

Figure 12.10 Principle of multiplex detection of miRNAs by the mixed probes. ()

Figure 12.11 Schematic illustration of the overall strategy for HAase detection. ()

Figure 12.12 Schematic representation of proposed sensing mechanism for the detection of surfactants SDS/SDBS and spermine. ()

Figure 12.13 As assay for the detection of ACP in human blood serum using assembly of WSCP P1 and Fe

3+

()

Figure 12.14 FRET‐assisted selective detection of bilirubin in human blood serum using water‐soluble conjugated polyfluorene PF–Ph–GlcA. ()

Figure 12.15 Basic mechanism of action of photodynamic therapy.

Figure 12.16 Mechanistic route of

1

O

2

generation by WSCPs by different pathways. ()

Figure 12.17 (a) Chemical structure of the PEGylated CPE covalently linked with anticancer drug doxorubicin (DOX) via a UV‐cleavable linker CPE–DOX and (b) schematic illustration of the near‐infrared (NIR) laser regulated initiation of the photosensitizer to generate ROS for photodynamic therapy and on‐demand drug release for chemotherapy ()

Figure 12.18 Schematic of the light‐activated hypoxia‐responsive drug‐delivery system. (a) Formation and mechanism of DOX/CP‐NI NPs. (b) Schematic of DOX/CP‐NI NPs for ROS generation and inducing a local hypoxic environment capable of hypoxia‐responsive release DOX into cell nuclei for enhanced synergistic anticancer efficacy ()

Figure 12.19 (a) Schematic illustration of PorCP NP formation; (b) DLS result of PorCP NPs in aqueous solution (the inset is the TEM image of PorCP NPs); and (c) UV–vis spectrum of PorCP NPs in aqueous medium. ()

Figure 12.20 Schematic illustration of PLL‐

g

‐PEG/DNQ (PFTTQ/DOX NPs) formation and the combination of photothermal therapy and chemotherapy regulated by a two‐photon laser ()

Figure 12.21 Chemical structures of different conjugated polyelectrolytes and fluorescence images of islet amyloid in human pancreas. (b) and (c) Tissue stained with PTAA (a; 2.5 µg ml

−1

) in Na carbonate (100 mM, pH 10) for 2 h. (e) and (f) Tissue stained with PONT (d; 2.5 µg ml

−1

) in glycine⋅HCl (100 mM, pH 2.5) for 2 h. (h) and (i) Tissue stained with POMT (g; 2.5 µg ml

−1

) in Na carbonate (100 mM, pH 10) for 2 h. (k) Tissue stained by Congo red (j). Scale bars (white lines) represent 150 µm. ()

Figure 12.22 LCP staining of sectioned brains from tg‐APP Swe mice at different ages. (a) Histological staining of hippocampus of an 18 mo tg‐APP Swe mouse with tPTT (405 nm filter). (b) Histological staining of hippocampus of a 8 mo tg‐APP Swe mouse with tPTT (405 nm filter) (c) Histological staining of a core plaque in the cerebral cortex of a 18 mo tg‐APP Swe mouse by PTAA (470/546 nm filters). The scale bars indicate 50 µm. ()

Figure 12.23 (a,b) High resolution fluorescence images showing an overview of the interplay between Aβ deposits (green), NFTs, and dystrophic neurites (yellow red). (c) Emission spectra of p‐FTAA bond to Aβ aggregates (green spectrum) or NFTs (red spectrum). (d,e) High resolution fluorescence images showing the details of the interplay between Aβ deposits (green), NFTs, and dystrophic neurites (yellow red). Selected Aβ deposits and NFTs are highlighted (green and red arrows, respectively) to indicate striking spacial co‐localization. Scale bar = 50 µm (image a), 20 µm (image b), and 10 µm (images d and e). ()

Figure 12.24 (a) PF‐HQ self‐aggregates in aqueous medium (b) modulation of amyloid aggregates in the presence of PF‐HQ and formation of polymer–protein coaggregates. ()

Chapter 13: Polymer‐Grafted Conjugated Polymers as Functional Biointerfaces

Figure 13.1 (a) Typical AFM height images of PEG‐grafted CPs as well as unfunctionalized CP surfaces. () (b) Frequency response demonstrating the relative protein resistance of ungrafted PBrEDOT, and PBrEDOT grafted with PDEGMMA and P(PEGMMA

‐co‐

DEGMMA, monomer ratio 1:9 brushes in serum‐free media and 20% serum, measured using Quartz Crystal Microbalance with Dissipation. () (c) Fibroblast adhesion on PBrEDOT grafted with P(PEGMMA‐

co

‐DEGMMA) brushes of different compositions, showing variable cell adhesion based on brush composition. Blue = DAPI (nucleus), red = phalloidin (actin). ()

Figure 13.2 (a) General biosensor design, where the interaction between the target analyte and the sensing element (recognition layer), such as CP‐based graft copolymer, is readout as an electronic signal. () (b) Various electrochemical techniques involved in characterizing the surface of graft copolymer modified electrodes. (Zig zag direction from top left: cyclic voltammetry, scan rate dependence of peak currents, and Nyquist diagrams obtained by electrochemical impedance spectroscopy) [103]. (c) Electrochemical detection of various analytes at the graft copolymer modified electrode surface. ()

Figure 13.3 (a–d) Optical microscope images of NIH3T3 fibroblast cells on oligoethylene glycol‐containing polythiophene surfaces at different culture times, demonstrating its use as biointerface for tissue engineering scaffolds. (e) Synthesis of polythiophene with oligoethylene glycol side chains. (f) Comparison of cell density of NIH3T3 fibroblast cells cultured on standard tissue culture plate (red) and on functional oligoethylene glycol‐containing polythiophene surfaces (rest of the color). (

Figure 13.4 (a) Thermoresponsive behavior of PNIPAAm‐grafted polythiophene, showing the change in conformation with temperature below or above its LCST. () (b) Graphical illustration showing the reversible helix‐coil transition of the grafted peptide side chains of a poly(phenylacetylene)‐based graft copolymer with the change in pH. () (c) Electrochemically responsive wettability and conformational switch of zwitterionic polymer brushes grafted from a polypyrrole‐based backbone. ()

Figure 13.5 Generalized categories of graft copolymer synthesis: grafting through (macromonomer approach), grafting to, and grafting from (macroinitiator approach).

Figure 13.6 Synthesis of poly(fluorene–phenylene)‐

g‐

(polystyrene) from a mid‐chain functionalized macromonomer. ()

Figure 13.7 (a) Synthesis of densely grafted PEG from a conjugated polymer through click chemistry. The graft copolymer was further modified with folic acid to fabricate molecular brush based cellular probe ( b, d). Fluorescence and (e, g) fluorescence/transmission overlapped images of MCF‐7 cells stained by graft copolymers with and without folic acid modification. (c) Fluorescence and (f) fluorescence/transmission overlapped images of unstained MCF‐7 cells. Strong fluorescence is shown for folic acid modified graft copolymer stained MCF‐7 cells (d, g). ()

Figure 13.8 Reaction scheme showing the interchangeable functionalization of CPs through “grafting from” ATRP as well as “grafting to” “click” reactions. ()

Figure 13.9 General scheme showing the electrodeposition of CTA‐containing thiophene monomer and carbazole, where polymer brushes can be grown through R‐group RAFT approach.

Guide

Cover

Table of Contents

Begin Reading

Begin Reading

Pages

xi

xii

xiii

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

80

81

82

83

84

85

87

88

89

90

91

92

93

95

96

97

98

99

101

103

105

106

107

108

109

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

135

136

137

138

139

140

141

142

144

145

146

148

150

152

153

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

182

183

184

185

186

187

188

189

190

191

192

193

194

195

196

197

199

201

202

203

204

205

206

207

209

210

211

212

213

215

216

218

219

220

221

222

224

225

226

228

229

230

231

232

234

235

236

237

238

239

240

241

243

244

245

246

247

248

249

250

251

252

253

254

255

256

257

258

259

260

261

262

263

264

265

266

267

269

270

271

272

273

274

277

279

280

281

282

283

284

285

286

287

288

289

290

291

292

293

294

295

296

297

298

300

301

303

304

305

306

307

309

312

313

316

317

318

319

320

321

322

323

324

325

326

327

329

330

331

333

334

335

336

337

338

339

340

341

342

343

344

345

346

347

348

349

350

351

352

353

354

355

356

357

358

359

360

361

362

363

365

366

367

369

370

371

372

373

374

375

376

377

378

381

382

383

385

386

387

388

389

390

391

392

393

394

395

396

397

398

399

400

401

403

404

405

406

407

408

409

410

Conjugated Polymers for Biological and Biomedical Applications

Edited by Bin Liu

Editor

Prof. Bin Liu

National University of Singapore

Chemical and Biomolecular Engineering

21 Lower Kent Ridge Road

Singapore 119077

Cover: © Kotkoa/Gettyimages

All books published by Wiley-VCH are carefully produced. Nevertheless, authors, editors, and publisher do not warrant the information contained in these books, including this book, to be free of errors. Readers are advised to keep in mind that statements, data, illustrations, procedural details or other items may inadvertently be inaccurate.

Library of Congress Card No.: applied for

British Library Cataloguing-in-Publication Data

A catalogue record for this book is available from the British Library.

Bibliographic information published by the Deutsche Nationalbibliothek

The Deutsche Nationalbibliothek lists this publication in the Deutsche Nationalbibliografie; detailed bibliographic data are available on the Internet at http://dnb.d-nb.de.

© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Boschstr. 12, 69469 Weinheim, Germany

All rights reserved (including those of translation into other languages). No part of this book may be reproduced in any form – by photoprinting, microfilm, or any other means – nor transmitted or translated into a machine language without written permission from the publishers. Registered names, trademarks, etc. used in this book, even when not specifically marked as such, are not to be considered unprotected by law.

Print ISBN: 978-3-527-34273-0

ePDF ISBN: 978-3-527-34278-5

ePub ISBN: 978-3-527-34275-4

Mobi ISBN: 978-3-527-34277-8

oBook ISBN: 978-3-527-34274-7

Preface

Since the conferring of the Nobel Prize in Chemistry to conducting polymers in 2000, this versatile class of polymeric materials has generated tremendous interest from academia and industry. Conjugated polymers are organic macromolecules with π‐conjugated backbone, which could be designed to display high electrical conductivity, outstanding photophysical properties, and excellent biocompatibility. They have been actively explored for a wide range of applications, spanning electronics, and energy harvesting to nanobiotechnology and nanomedicine. While the last 15 years have witnessed their rapid development in optoelectronic devices, the emerging field of conjugated polymers for biomedical applications has simultaneously attracted increasing attention recently. Specifically, they have found innovative applications in a variety of biotechnologies, ranging from biosensing that takes advantage of optical amplification, to cell imaging and image‐guided therapy that fully utilizes the light‐harvesting properties of conjugated polymers and their structure‐dependent radiative and nonradiative pathways as well as their self‐assembly properties with amphiphilic molecules. In fact, with their unique biophysicochemical attributes, conjugated polymer nanoparticles have been demonstrated to be even superior to other classes of nanomaterials, such as small organic dye‐based nanoparticles, semiconducting quantum dots, and inorganic nanoparticles, in many biomedical applications. Remarkably, with the continual advancement in polymer design, synthesis, and processing strategies in the last few years, conjugated polymers have been increasingly reported to be highly promising for bioimaging, sensing, and for applications in disease therapy.