Table of Contents

Title Page

List of Contributors

Chapter 1: Pharmacokinetics and Pharmacodynamics (PK/PD) of Bionanomaterials

1.1 Introduction
1.2 Commonly Utilized NMs in Pharmaceutical Research
1.3 In vivo Biodistribution and the Evolving Targeting Principles for NMs
1.4 Processing NMs by the Biological Systems
1.5 Rational Design of Long-Circulating NMs
1.6 Mathematic Simulation of NM-Mediated Cancer Drug Delivery
1.7 Experimental PK Data of the Applied NMs
1.8 Perspectives
References

Chapter 2: Targeted Dendrimers for Cancer Diagnosis and Therapy

2.1 Introduction
2.2 Targeted Dendrimers for Cancer Therapy
2.3 Targeted Dendrimers for Cancer Diagnosis
2.4 Conclusions
References

Chapter 3: Polymeric Micelles for Drug Delivery

3.1 Introduction
3.2 Amphiphilic Copolymers for Micelle Preparation
3.3 Stability of Polymeric Micelles
3.4 Drug Incorporation of Polymeric Micelles
3.5 Functionalization of Polymeric Micelles
3.6 Conclusions
References

Chapter 4: Polymeric Micelle-Based Nanomedicine

4.1 Introduction to Chemotherapy
4.2 Polymeric Micelle-Based Nanomedicine
4.3 Perspective
References

Chapter 5: Microfluidics Applications in Cancer Drug Delivery

5.1 Introduction
5.2 Basic Principles of Micellar Drug Carriers and Microfluidics
5.3 Microfluidic Fabrication of Polymer Micelles
5.4 On-Chip Characterization of Micelle Formation
5.5 Microfluidic Replications of Physiological Barriers During Delivery of Drug to Tumor
5.6 Conclusion and Implications for Future Research
Acknowledgment
References

Chapter 6: Antibody–Drug Conjugates

6.1 Introduction
6.2 History of ADCs
6.3 Components of ADCs
6.4 Future Directions
References

Chapter 7: Nano-Photosensitizer for Imaging-Guided Tumor Phototherapy

7.1 Introduction for Tumor Phototherapy
7.2 Functionalized Nano-Photosensitizer for Tumor Targeting
7.3 Nano-photosensitizer for Photodynamic Therapy
7.4 Nano-Photosensitizer for Photothermal Therapy
7.5 Nano-Photosensitizer for Combination Therapy
7.6 Perspective and Application
References

Chapter 8: Quantum Dots for Cancer Diagnosis

8.1 Introduction
8.2 Detection of Solid Tumor Based on QDs
8.3 SLN Mapping
8.4 Detection of Tumor-Associated Proteins in Blood
8.5 Detection of CTCs
8.6 Tumor Microenvironment for Invasion and Metastasis
8.7 Challenges of QDs into Clinical Practice Application
8.8 Summary
References

Chapter 9: Luminescent Gold Nanoclusters for Biomedical Diagnosis

9.1 Gold Nanostructures in Biomedical Diagnosis
9.2 Luminescent Au NCs for Biosensing
9.3 Au NCs for Cell Imaging
9.4 Au NCs for In Vivo Imaging
9.5 Perspectives
References

Chapter 10: Nanographene in Biomedical Applications

10.1 Introduction
10.2 Nanographene for Drug Delivery
10.3 Nanographene for Gene Delivery
10.4 Graphene-Based Nanocomposite for Drug Delivery
10.5 Nanographene for Phototherapies of Cancer
10.6 Graphene and its Nanocomposites for Biomedical Imaging and Imaging-Guided Therapy
10.7 Toxicity of Nanographene
10.8 Prospects and Challenges
References

Chapter 11: Molecular Imprinting Technique for Biomimetic Sensing and Diagnostics

11.1 Introduction
11.2 Molecularly Imprinted Polymers (MIPs)
11.3 MIPs for Biomimetic Sensing and Diagnostics
11.4 Conclusions and Outlook
Acknowledgments
References

Chapter 12: Magnetic Nanostructures for MRI-Based Cancer Detection

12.1 Introduction
12.2 Chemical Synthesis of Magnetic Nanostructures
12.3 Magnetic Nanostructures for MRI-Based Cancer Detection
12.4 Conclusions and Perspective
Acknowledgments
References

Chapter 13: Magnetic Iron Oxide Nanoparticles: Bioapplications and Potential Toxicity

13.1 Introduction
13.2 Bioapplications of Magnetic Iron Oxide Nanoparticles
13.3 Potential Toxicity of Magnetic Iron Oxide Nanoparticles
13.4 Surface Engineering for Bioapplications
13.5 Conclusion
Acknowledgments
References

Chapter 14: Nanostructured Hydrogels for Diabetic Management

14.1 Introduction
14.2 Nanostructured Hydrogels for Insulin Releasing
14.3 Nanostructured Hydrogels for Glucose Sensing
14.4 Nanostructured Hydrogels in Artificial Pancreas
14.5 Conclusions and Outlook
References

Chapter 15: Inorganic Nanomaterials for Bone Tissue Engineering

15.1 Introduction
15.2 Calcium Phosphate Nanomaterials for Bone Tissue Engineering
15.3 CaP Blocks and Scaffolds with Surface Nanostructure
15.4 Mesoporous Bioactive Glasses for Bone Tissue Engineering
15.5 Conclusions
Acknowledgments
References

Chapter 16: Nanotechnology in Coronary Artery Stent Coating

16.1 Introduction
16.2 Biodegradable Polymer Coating
16.3 Nanocomposite Stent Coating
16.4 Nanostructure in Stent Coating
16.5 Bioactive Nanocoating
16.6 Summary and Future Outlook
References

Index

End User License Agreement

List of Illustrations

Chapter 1: Pharmacokinetics and Pharmacodynamics (PK/PD) of Bionanomaterials

Figure 1.1 Schematic illustration of lipid-based NMs.
Figure 1.2 Various forms of PEI.
Figure 1.3 Schematic illustrations of activatable nanoprobe. The nanoprobe comprises two components (the fluorescence donor QD-LH and the FRET quencher QSY21), which are cross-linked via a chimeric peptide comprising LP and a legumain-cleavable sequence [60].
Figure 1.4 Schematic illustration of folate-modified pH-sensitive PAMAM.
Figure 1.5 Distribution of blood flow in the pulmonary and systemic circulations [81].
Figure 1.6 The tissue-specific extravasation of NMs [84].
Figure 1.7 Schematic illustrations of one- and two-compartment models.
Figure 1.8 Pharmacokinetic model for free and liposomal DOX [105]. Open and closed circles represent free and liposomal DOX, respectively. Liposomal DOX in CAP was transported into INT unidirectionally (k_tu). Liposomal DOX was taken up by RES (k_res) and efflux of DOX was neglected. Rapid equilibrium was assumed between CAP and INT for free DOX and C_ecs was defined for free DOX in the extracellular compartment (ECS). Distribution of free DOX to tumor cells was described using k_et and k_te. The parameters k₁₂, k₂₁, and k₁₀ represent the micro-pharmacokinetic constants for free DOX.
Figure 1.9 Changes of C_max and T_max in different compartments with different K_rel and K_res. The parameters C_{int lipo}, C_{ecs dox}, and C_{tu dox} denote liposomal dox in the interstitial space, free dox in the ECS, and free dox in the tumor site, respectively; T_{int lipo}, T_{ecs dox}, and T_{tu dox}, the time at which C_max was reached.
Figure 1.10 Simulations of the time courses of free and liposomal DOX in mice bearing P388 tumor in the peritoneal cavity [106]. With the other fixed parameters, while k_rel value was adjusted at: (a) k_rel = 0.006 (h⁻¹), (b) k_rel = 0.06 (h⁻¹), (c) k_rel = 0.6 (h⁻¹), and (d) k_rel = 6 (h⁻¹).
Figure 1.11 Effect of k_rel on the antitumor effect of liposomal DOX [106].
Figure 1.12 (a) A typical blood flow-limited physiologically based pharmacokinetic model structure; (b) a membrane-limited tissue; and (c) a blood flow-limited tissue [108].
Figure 1.13 Schematic representation of drug RADME process in a polymer formulation [111].
Figure 1.14 Comparison of simulated vinpocetine concentration with experimental data both in vitro (a, showing C_r–time profile in GI) and in vivo (b, showing C_b–time profile in blood). In both sets, solid lines refer to the simulated data, while open or filled circles represent experimental data from different tests [111].
Figure 1.15 Organ content of triphenylphosphine-coated AuNP with different size.

Chapter 2: Targeted Dendrimers for Cancer Diagnosis and Therapy

Figure 2.1 Two strategies in the targeted delivery of anticancer drugs to tumor. (a) Passive targeting using the EPR effect of nanoparticles and (b) active targeting via ligand–receptor recognition.
Figure 2.2 Chemical structures of PAMAM and PPI dendrimers.
Figure 2.3 Structures of FA-targeted dendrimer therapeutics. (a) FA-, MTX-, and FITC-modified dendrimer, (b) triple-functionalized dendrimer synthesized by copper-free click chemistry, (c) FA-targeted dendrimer for boron neutron capture therapy, (d) FA-targeted hybrid nanocluster linked by complementary oligonucleotides, and (e) FA-targeted hybrid nanocluster linked by click chemistry.
Figure 2.4 Anticancer drugs are encapsulated within (a) or covalently conjugated to (b) FA-modified dendrimers for targeted cancer therapy.
Figure 2.5 Structures of lactose-, biotin-, riboflavin-, and estrogen-modified dendrimers for targeted cancer therapy.
Figure 2.6 Targeting cancer cells using a three-step strategy based on specific biotin–avidin recognition.
Figure 2.7 HA-modified dendrimers in the codelivery of DOX and MVP siRNA to specific tumors.
Figure 2.8 Targeted dendrimers in cancer diagnosis. (a) Targeted dendrimer encapsulated with gold nanoparticles, (b) targeted dendrimer conjugated with DTPA/^99mTc, (c) targeted dendrimer conjugated with DTPA/Gd³⁺, and (d) targeted dendrimer conjugated with Cy5.5.
Figure 2.9 Advantages and drawbacks of different imaging tools in targeted cancer diagnosis.
Figure 2.10 Capture of CTCs on a substrate surface immobilized with (a) G7-aEpCAM_4.9 and (b) PEGylated.

Chapter 3: Polymeric Micelles for Drug Delivery

Figure 3.1 Schematic diagram for the formation and disassociation of polymeric micelles prepared from amphiphilic diblock copolymers (CMC, critical micelle concentration).
Figure 3.2 Typical hydrophilic polymers with excellent anti-biofouling ability.

Chapter 5: Microfluidics Applications in Cancer Drug Delivery

Figure 5.1 Synthesis, characterization, and in vitro evaluation of polymeric micelle drug carriers in microfluidic devices [12].
Figure 5.2 Polymeric micelle formation [2].
Figure 5.3 (a) Diffusive mixing [56]; (b) herringbone mixing [58]; (c) Tesla mixing [59]; (d) microvortex mixing [60]; (e) droplet-based mixing [61].
Figure 5.4 Integration of QD-FRET and microfluidics to monitor polyplex self-assembly kinetics in a simple microfluidic system [73].
Figure 5.5 Schematic diagram of integration of Raman spectroscopy and small-angle X-ray scattering in microfluidic device [95].
Figure 5.6 β-Carotene/Pluronic F127 hybrid NPs fabricated in three-inlet microreactor via HFF [99].
Figure 5.7 Schematic diagram of microfluidic system integrating ATRP synthesis of diblock copolymers, micellization, and in situ nanoscale particle sizing with DLS [102].
Figure 5.8 Mechanically sensitive drug delivery system: release of drug component physically triggered by change in shear stress at clogged artery [139].
Figure 5.9 Development of endothelialized microfluidic device to probe nanoparticle translocation over permeable microvessel [143].
Figure 5.10 Fluorescent PEG NPs (40 nm) entering spheroid-immobilized microfluidic device and accumulating in spheroid interstitial spaces [152].

Chapter 6: Antibody–Drug Conjugates

Figure 6.1 Antibody–drug conjugate (ADC) timeline [25].
Figure 6.2 Structures of (a) BR96-DOX (anti-BR96 doxorubicin conjugate) and (b) Mylotarg® (anti-CD33 calicheamicin conjugate).
Figure 6.3 Structure of Adcetris® (anti-CD30 auristatin conjugate).
Figure 6.4 Structure of Kadcyla® (anti-Her2 maytansine conjugate).
Figure 6.5 Schematic diagram of ADC components [25].
Figure 6.6 Structure of common drugs of ADCs.
Figure 6.7 Mechanism of action of ADCs.
Figure 6.8 Random conjugation and site-specific conjugation strategies in ADCs [25].

Chapter 7: Nano-Photosensitizer for Imaging-Guided Tumor Phototherapy

Figure 7.1 ICG-loaded lipid–polymer hybrid nanoparticles (INPs) with different sizes for MCF-7 breast adenocarcinoma tumor PTT. (a) TEM images of INP1, INP-2, and INP-3 (scale bar = 50 nm); (b) temperature-increasing profiles in BxPC-3 tumor tissue after in vivo photothermal treatment (808 nm, 0.8 W cm⁻², 10 min), 24 h after intravenous injection of the INPs; (c) the growth curve of BxPC-3 xenograft tumors within 4 weeks in different groups after the treatments as indicated (n = 5); (d) histological staining of removed tumors at 48 h after injection of PBS, free ICG, INP-1, INP-2, or INP-3 plus laser irradiation (scale bar = 50 µm).
Figure 7.2 Protein-based, reduced graphene oxide (nano-rGO) for tumor theranostics. (a) A scheme showing the preparation of nano-rGO from nano-GO using BSA as reductant and stabilizer; (b) photoacoustic (PA) signals (across the center of the three phantoms) and PA images of nano-rGO (0.05 mg mL⁻¹), nano-GO (0.05 mg mL⁻¹), and agarose gel; (c) ultrasound and PA dual-modality images of the tumor region using nano-rGO as contrast agent; (d) histological staining of the excised tumors (bar = 50 µm) (808 nm, 0.6 W cm⁻², 5 min).
Figure 7.3 Schematic structures of DOX and ICG co-encapsulated nanoparticles (DINPs) and combined photothermal/chemotherapy in vivo. (a) Schematic illustration of the single-step sonication to synthesize DINPs. Photograph of mixture containing DOX, ICG, lecithin, and DSPE-PEG was before (left) and after (right) sonication. (b) Histological staining of the excised tumors 12 h after injection of PBS, free ICG, INPs, and DINPs under laser irradiation. Common features of thermal damage were identified in tumors treated with free ICG, INPs, and DINPs, such as coagulative necrosis (black arrow), abundant pykonosis (blue arrow), and considerable regions of karyolysis (red arrow), and cellular apoptosis of MCF-7 cells in tumor tissues induced by PBS plus laser, free ICG plus laser, free DOX, and DINPs plus laser. The apoptotic cells labeled green FL were evidently identified by TUNEL assay (scale bar, 50 µm). The tumor cells treated with ICG plus laser or DOX provided a great number of apoptosis signals because both hyperthermia and DOX could cause cell apoptosis. (c) MCF-7 and MCF-7/ADR tumor growth curves of different groups after treatments.
Figure 7.4 (a) Schematic illustration of HSA-ICG NPs for in vivo dual-modal imaging, tumor margin detection, and simultaneous PDT/PTT treatments. Upon the single NIR laser irradiation, the HSA–ICG NPs can simultaneously convert the absorbed light energy to ROS and heat for synergistic PDT/PTT treatments. (b) FL image of 4T1 cells after PDT, PTT, and simultaneous PDT/PTT treatments. Viable cells were stained green with calcein-AM, and dead/later apoptosis cells were stained red with PI. (c) In vivo tumor margin detection. The tumor, tumor margin, and normal tissue could be detected using in vivo NIR and PA dual-modal imaging and spectrum-resolved technology. (d) Tumor growth curves of different groups of 4T1 orthotropic mice.

Chapter 8: Quantum Dots for Cancer Diagnosis

Figure 8.1 QD-based in situ molecular imaging and multispectral analyses of tumor tissue specimen. (a) Tumor slides from gastric cancer specimens were first stained with two primary antibodies against macrophages and CD105, a marker of tumor neovessels, and then stained with QD-conjugated secondary antibodies, with the macrophages stained green (525-nm spectrum) and neovessels stained red (655-nm spectrum). (b, c) The images were computer-captured and unmixed by multispectral analysis software, to delete the signal noise and set the spectral images of macrophages and neovessels for subsequent analysis. (d) Quantitative analysis of tumor microenvironment with the computer-aided algorithm and the results output.
Figure 8.2 QD-based immunohistochemistry (IHC) labeled the molecular pathology. (a) QD protocol for single (a1) and multiple (a2) biomarker detection in situ. (b) HER2 staining through conventional IHC (b1), gene amplification through FISH (b2), and in situ imaging of HER2 (b3) and ER (b4) through QD-IHC. (c) BC heterogeneity indicated by single HER2 quantitative detection (c1) and double-color in situ imaging of HER2 and ER (c2), as well as cytokeratin and proliferating cell nuclear antigen (c3). (d) QD-based molecular classification of breast cancer by quantitative HER2 and hormone receptor information could differentiate subtypes with different 5-year disease-free survival to formulate more individualized therapy, including three subtypes based on total HER2 load (d1), three subtypes by hormone receptors (d2), five subtypes of the combination of total HER2 load and hormone receptors (d3), and 5-year recurrence risk of these five subtypes (d4). HHER2, high HER2 load; HHR, high hormone receptor; LHER2, low HER2 load; LHR, low hormone receptor; NHR, negative hormone receptor.
Figure 8.3 In vivo targeting and imaging of a lung metastasis model with QD-based nanotechnology. (a) The imaging system for living animal models. (b) In vivo targeted imaging of the subcutaneous tumor model, and the site-by-site spectrum analysis of the tumor, which showed that the QD-labeled anti-α-fetoprotein monoclonal antibody probes per field were lower in the center than in the periphery of the tumor, indicating that tumor growth was not homogeneous and the peripheral site was more active. (c) In vivo targeted imaging of liver cancer lung metastasis models.
Figure 8.4 Flow diagram of CTC detection system with magnetic nanospheres (MNs). Under this detection system, the MNs were nanosized with fast magnetic response, and nearly all of the MNs could be captured by 1-min attraction with a commercial magnetic scaffold. In addition, the MNs were very stable without aggregation or precipitation in whole blood and could be re-collected nearly at 100% in a monodisperse state. Modified with antiepithelial cell-adhesion molecule antibody, the obtained immunomagnetic nanospheres successfully captured extremely rare tumor cells in whole blood with an efficiency of more than 94% via only 5-min incubation. Moreover, the isolated cells could be directly used for culture, reverse transcription-polymerase chain reaction, and immunocytochemistry identification.
Figure 8.5 Six typical examples of negative correlation between the number of tumor invasion units and the OS of gastric cancer patients. In three cases of poorly differentiated adenocarcinoma (a, b, and c) and another three cases of highly differentiated adenocarcinoma (d, e, and f) of the same TNM stage and treatments, tumor invasion unit was negatively correlated with OS of each patient. The two graphs on the right-hand side represent the number of tumor invasion unit and the corresponding OS of each patient. Scale bar: 50 mm for a1-f4; OS:,overall survival (months); the number of tumor invasion unit was the count per 200 magnification field.

Chapter 9: Luminescent Gold Nanoclusters for Biomedical Diagnosis

Figure 9.1 (a, c) Bar diagram showing changes in the luminescence intensity of Au NCs: in the presence of different metal ions (a) and at various pH (c). (b, d) Corresponding images of the solutions in UV light.
Figure 9.2 Confocal images of mixed population of E. coli (left side of the visual field) and S. aureus (right side of the visual field) incubated with Au NCs probe: (a) phase contrast image and (b) fluorescence image of the same region. Scale bar is 10 µm.
Figure 9.3 z-Sectioning of SH-SY5Y neuroblastoma cell images obtained using Au NCs as two-photon fluorescence contrast agents under excitation of 800-nm femtosecond laser pulses.
Figure 9.4 Typical FLIM images of HeLa cells with internalized AuNCs at four different temperatures.
Figure 9.5 FLIM and time-gated intensity images of BSA Au NCs, fluorescein and mixture of BSA Au cluster and fluorescein-treated breast cancer cell line (4T1).
Figure 9.6 Microscopic imaging of insulin-Au NCs uptake by differentiated C2C12 myoblasts after 2-h treatment. (a) Cell nucleus stained with 4′,6-diamidino-2-phenylindole (DAPI, blue). (b) Actin fiber stained with Alexa Fluor 488 phalloidin to confirm the cell boundary (green). (c) Insulin-Au NCs (red). (d) Overlay of the three images.
Figure 9.7 Representative xenograft tumor mouse models of hepatocellular carcinoma observed in normal light (a) or by in vivo fluorescence imaging (b) 24 h after a subcutaneous injection of 10 mM HAuCl₄ solution near the tumor. In (a), the inset shows an enlarged view of the xenograft tumor. Xenograft tumor mouse models of chronic myeloid leukemia observed by in vivo fluorescence imaging (c) 24 h after a subcutaneous injection of 10 mM HAuCl₄ solution near the tumor. (d) Control mouse observed by in vivo fluorescence imaging 48 h after a subcutaneous injection of 10 mmol L⁻¹ HAuCl₄ solution in the right-hand side of their abdomen.
Figure 9.8 (a) NIR fluorescent blood imaging in vivo using BSA–Gd₂O₃/Au nanoprobe. (b) MR blood imaging in vivo using BSA–Gd2O3/Au nanoprobe.
Figure 9.9 In vivo 3D CT images of saline and BSA–Au cluster-injected mice at 2 h post injection in the (a) supine and (b) prone positions. (c) An anatomical diagram of the kidney. (d) An enlarged view of the major organs of the BSA–Au cluster-injected mouse (bone is subtracted out).

Chapter 10: Nanographene in Biomedical Applications

Figure 10.1 Schematic of functionalized nanographene for loading of small therapeutic molecules including DOX, SN38, and Ce6.
Figure 10.2 Schematic of functionalized nanographene for gene transfection.
Figure 10.3 Photothermally enhanced gene transfection. (a) Schematic of preparation of nGO–PEG–PEI and nGO–PEG–PEI/pDNA complex (b) The sizes change of nGO–PEG–PEI and GO–PEI incubated with in water or DMEM containing 10% fetal bovine serum (FBS). The inset shows an image of nGO–PEG–PEI (right) and GO–PEI (left) in complete cell medium after being centrifuged at 1000 rpm for 10 min. (c) Relative viability of HeLa cells treated with nGO–PEG–PEI, GO–PEI and PEI for 24 h. (d) Relative transfection efficiencies (TE) of nGO–PEP–PEI, GO–PEI, and bare PEI at different FBS concentrations. (e) Schematic of photothermally enhanced gene delivery. (f) Confocal fluorescence images of EGFP-transfected HeLa cells treated with nGO–PEG–PEI at different N/P ratios and culture temperature together with the 808-nm laser irradiation (0.5 W cm⁻²). (g, h). The expression levels of Plk1 mRNA (g) and protein (h) as determined by qRT–PCR and Western blotting, respectively.
Figure 10.4 Multifunctional nanographene-based targeted drug delivery. (a) Schematic of GSPID (DOX-loaded mesoporous silica-coated graphene nanosheet conjugated with targeting peptide (GSPI)) for combined chemophotothermal targeted therapy of glioma. (b) Confocal imaging of cells stained with LIVE–DEAD after different treatments. (c) Relative viability of glioma cells after different treatments.
Figure 10.5 Optimization of graphene-based in vivo photothermal therapy. (a) AFM images of three GO derivatives. nGO–PEG and nRGO–PEG showed similar sizes of about 20–30 nm, while the size of RGO–PEG was about 60–70 nm. In addition, compared to nGO–PEG with covalent PEG conjugation, noncovalently functionalized nRGO–PEG and RGO–PEG showed increased sheet thickness, likely owing to their more condensed PEG coatings. (b) Blood circulation of GO derivatives. The blood from the mice intravenously injected with ¹²⁵I-labeled nGO–PEG, nRGO–PEG, and RGO–PEG was collected at various time points after treatment and measured by gamma counter (n = 3). (c) Biodistribution of GO derivatives in 4T1 tumor-bearing mice 2 days after injection. (d) 4T1 tumor growth curves of mice after various treatments indicated. The laser irradiation was carried out at the power density of 0.15 W cm⁻² for 5 min. (e) Survival of mice bearing 4T1 tumors after various treatments indicated.
Figure 10.6 Preparation of pH-responsive nanographene. (a) Schematic of the acidic extracellular environment-induced charge reverse of fabricated nGO–PEG–DA/DOX complex. (b) Zeta potential changes of nGO–PEG–DA and nGO–PEG–SA incubated in the PBS of pH 6.8 or 7.4. (c) Relative viabilities of MCF-7/ADR cells incubated with nGO–PEG–DA/DOX and free DOX at the DOX concentration of 20 µg mL⁻¹ and nGO–PEG–DA at the equivalent nGO concentration, with or without being irradiated by the 808-nm laser for 5 min at the power density of 0.5 W cm⁻² after 48-h incubation.
Figure 10.7 In vitro and in vivo fluorescence imaging using functionalized nanographene. (a) Scheme showing anti-CD20 antibody-conjugated nGO–PEG to target Raji B cells. (b) Photoluminescence images of CD-20-positive Raji B cells and CD-20-negative CEM cells incubated with anti-CD-20-conjugated nGO–PEG. (c) A scheme of nGO–PEG labeled with Cy7. (d) In vivo fluorescence images of mice bearing 4T1, KB, and U87MG tumors taken at different time points after i.v. injection of nGO–PEG–Cy7.
Figure 10.8 In vivo PET imaging of radiolabeled nGO–PEG. (a) Scheme of nGO–PEG conjugated with anti-CD105 antibody (TRC105) and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). The latter was used for ⁶⁴Cu labeling. (b) Time-dependent in vivo PET imaging of mice bearing 4T1 tumors i.v. injected with ⁶⁴Cu–NOTA–nGO–TRC105, ⁶⁴Cu–NOTA–nGO, and ⁶⁴Cu–NOTA–nGO–TRC105 after a preinjected blocking dose of TRC105. Tumors are indicated by arrowheads.
Figure 10.9 Multimodal imaging and imaging-guided photothermal therapy using PEGylated RGO–IONP nanocomposite (RGO–IONP–PEG). (a) A scheme showing preparation and functionalization of RGO–IONP. (b, c) TEM images of as-prepared RGO–IONP (b) and RGO–IONP–PEG (c). Inset showing a high-resolution TEM image of an IONP grown on the RGO sheet. (d) Multimodal imaging including fluorescence, MR, and photoacoustic imaging of 4T1-tumor-bearing mice using RGO–IONP–PEG as the contrast agent. (e) Tumor-bearing mice before and after photothermal therapy with RGO–IONP–PEG injection and laser irradiation (808 nm, 0.5 W cm⁻², 5 min). (f) Monitoring of the therapeutic response by MR imaging of RGO–IONP–PEG injected mice with or without laser irradiation.
Figure 10.10 Signaling pathway of graphene induces cell apoptosis.
Figure 10.11 In vivo biodistribution and toxicity study of nGO–PEG. (a) Scheme of nGO–PEG labeled with ¹²⁵I. (b) Long-term biodistribution of mice intravenously injected with ¹²⁵I–nGO–PEG at different time points. (c–e) H&E stained liver slices collected from control untreated mice (c), mice treated with GO–PEG 3 days (d), and 20 days (e) post injection. Black dots pointed out by arrows were aggregated nGO–PEG in the liver. (f) Imaging of H&E stained slices of organs from the control group and nGO–PEG-treated group at different time points. No obvious organ damage was observed in 90 days.
Figure 10.12 Pulmonary toxicity induced by different forms of graphene. Mice were treated with Pluronic dispersed graphene (GD), aggregated graphene (GA), and GO by intratracheal instillation. The lungs were examined 21 days after treatment. (a) Photomicrographs of paraffin blocks of the lung after sectioning. (b–d) Photomicrographs of lung sections at 1× (b), 50× (c), and 200× (d). (e) The percentage of TUNEL-positive nuclei in paraffin-embedded lung sections. (f) Total lung collagen determined by picrosirius red precipitation of whole lung homogenates. (g and h) In vitro data showing generation of ROS (g) and the resulted DNA fragmentation (h) in an alveolar macrophage cell line. Water (−) or water with 2% Pluronic (−)P were used as negative controls.

Chapter 11: Molecular Imprinting Technique for Biomimetic Sensing and Diagnostics

Figure 11.1 Schematic representation of the generation of molecularly imprinted polymers.
Figure 11.2 Schematic representation for the preparation of the AuNPs@ MIES (MIP-modified sensor).
Figure 11.3 Preparation of the catalytic MIP-hybrid electrodes imprinted with catechol.
Figure 11.4 Chemical structures of the fluorescent functional monomers used for the synthesis of fluorescent MIPs.
Figure 11.5 (a) Microtip fabrication process. (b) Analysis by spectrofluorimetry of the polymer microtips with a simple fiber, and (c) a bifurcated fiber. A 375-nm diode laser is used both for microtip polymerization and excitation of the fluorophore.
Figure 11.6 (Above) Chemical structure of (a) MABA as a functional monomer and (b) p-isothiocyanatophenyl α-d-mannopyranoside (MITC) as a capping agent. (Below) Schematic drawing of the two-step postimprinting modification process. (a) First postimprinting modification (capping treatment by MITC); (b) second postimprinting modification (site-directed FITC introduction).
Figure 11.7 Self-assembly of the QDs, the polymerizable surfactant OVDAC, and boronic acid and acryamide monomers for the preparation of a fluorescence nanosensor.
Figure 11.8 (a) Electropolymerization of a bis-aniline-cross-linked AuNP composite for the sensing of TNT by donor–acceptor interactions; (b) imprinting of TNT molecular recognition sites into the bis-aniline-cross-linked AuNP composite associated with a Au electrode.
Figure 11.9 Scanning electron micrographs of (a) polystyrene bead-coated surface (100-nm spheres), and electrochemically deposited biotin-imprinted polymer films grown in the presence of (b) 100-nm, (c) 300-nm, and (d) 800-nm diameter sacrificial bead layered Au/quartz surfaces. An MIP film prepared in the absence of polystyrene beads is shown in (e). (f) A surface with visible underlying interconnected pore structure (prepared with 800-nm diameter sacrificial beads).
Figure 11.10 (Left) Schematic of the preparation of the protein surface-imprinted polymer coating on a QCM electrode. A stamp with densely packed rubber proteins is pressed into the prepolymerized coating, templates are removed after polymerization; (right) Topographic AFM images of poly(MAA–NVP–DHEBA–polymer) layer: (a) Hev b1 (L) and (b) Hev b1 (G) self-assembled on the glass surface, (c) Hev b1 (L)-MIP, and (d) Hev b1 (G)-MIP coated on a QCM electrode showing nanopatterned surfaces with nanoscale of lithographic imprints. Layer heights are 200 nm (spin-coated, layer height-tested with AFM).

Chapter 12: Magnetic Nanostructures for MRI-Based Cancer Detection

Figure 12.1 (a) Schematic illustration of the two environment-dependent growth routes (nucleation in two different environments). The blue area indicates an amine-rich environment, which represents an organic shell composition that preferentially stabilizes the {100} facets of the iron NCs. (Reproduced with permission from [42]. Copyright 2009, American Chemical Society.)(b) Mössbauer spectra of the iron NPs prepared after different reaction time and temperature.
Figure 12.2 (a) TEM images of Fe/Fe₃O₄ nanoparticles. (Inset) HRTEM image of the Fe/Fe₃O₄ nanoparticles. (Reproduced with permission from [46]. Copyright 2006, American Chemical Society.)(b) TEM image of the bcc-Fe NCs obtained from the redispersion of the plate assembly in hexanes. (Inset) HRTEM image of the Fe/Fe₃O₄ nanoparticles.
Figure 12.3 TEM images of FePt nanowires and nanorods.
Figure 12.4 (a) 3D representation of the reconstructed volume along different viewing directions of the Fe–Co dumbbell. (b) HAADF-STEM image of the particle focusing on the Fe cube. (c) M–H hysteresis loops at 2 K for Co nanorods (black), Fe nanocubes (blue), and Fe–Co dumbbell (red). (d) Simulated M–H hysteresis loops along the nanorod axis for a single Co nanorod (black), a single Fe nanocube along the $c12-math-0001$ direction (blue), and a single Fe–Co dumbbell along the long axis (red) using the OOMMF micromagnetic code at 0 K.
Figure 12.5 (a) Schematic illustration of the synthesis of monodispersed Fe₃O₄ NPs through the decomposition of Fe(acac)₃ in the presence of oleic acid, oleylamine, and alkane diol. Transmission electron microscopy (TEM) image of (b) 6-nm and (c) 10-nm Fe₃O₄ NPs. (d) Three-dimensional (3D) superlattice of 10-nm Fe₃O₄ NPs.
Figure 12.6 (a) A protocol for Fe₃O₄ nano-octahedra synthesis. (b) TEM image of octahedral Fe₃O₄ NPs with projection axis {111}.
Figure 12.7 TEM images of FeO NPs with different sizes and shapes: (a) 14-nm spherical, (b) 32-nm, and (c) 53-nm truncated octahedral. (d) Scanning electron microscopy (SEM) image of 100-nm truncated octahedral NPs.
Figure 12.8 TEM images of 14-nm (a) CoFe₂O₄ NPs and (b) MnFe₂O₄ NCs. (Reproduced with permission from [86]. Copyright 2004, American Chemical Society.)(c) TEM image, and (d) high-resolution TEM (HRTEM) image of 15-nm (Zn_0.4Fe_0.6) Fe₂O₄ NPs.
Figure 12.9 (a) Wrap–bake–peel process to obtain nanocapsules from akagenite. (Reproduced with permission from [96]. Copyright 2008, Nature Publishing Group.)(b) Synthesis of core–shell–void Fe–Fe₃O₄ and hollow Fe₃O₄ NPs from Fe–Fe₃O₄ NP seeds.
Figure 12.10 (a) TEM image of 20-nm iron carbide NCs. (b) Schematic illustration of the formation mechanism of Fe₅C₂ NPs. (Reproduced with permission from [4]. Copyright 2012, American Chemical Society.)(c) TEM image of 13.1-nm iron carbide NCs.
Figure 12.11 MR contrast effect of octapod iron oxide NPs. (a) Schematic shows the ball models of octapod and spherical iron oxide NPs with the same geometric volume (black dotted lines represent the magnetic field of the octapod and spherical iron oxide NPs. The same length of black arrow indicates the same Ms of octapod and spherical iron oxide NPs). With the same geometric core volume, the octapod NPs have much larger effective volume (radius, R) than the spherical nanoparticles (radius, r) with R about 2.4r under an external magnetic field B₀. (b) The smooth M–H curves of Octapod-30, Octapod-20, Spherical-16, and Spherical-10 measured at 300 K using a superconducting quantum interference device magnetometer (inset: M–H curves of Octapod-30 and Octapod-20 in low-magnetic field areas). The M values of Octapod-30, Octapod-20, Spherical-16, and Spherical-10 are about 71, 51, 67, and 55 emu g⁻¹, respectively. (c) T₂-weighted MR images of Octapod-30, Octapod-20, Spherical-16, and Spherical-10 in aqueous solution with 1% agarose at various concentrations of iron using a Varian 7-T micro-MRI scanner. (d) Comparison of r₂ values of Octapod-30, Octapod-20, Spherical-16, and Spherical-10. The error bars represent ±s.d. of five independent experiments.
Figure 12.12 MR contrast effect of ferrimagnetic iron oxide nanoparticles on changes in size. (a) T₂-weighted MR images of ferrimagnetic iron oxide nanoparticles at various concentrations of iron at 3 T and (b) their color-coded images. (c) Plots of R₂ values of ferrimagnetic iron oxide nanoparticles and (d) comparison of their r₂ values.
Figure 12.13 MR measurements and imaging of functionalized FeCo/GC NPs, Feridex and Magnevist solutions. (a) r₁ and (b) r₂ for the various solutions. (c) MR images of various contrast agents at three metal concentrations generated on a T₂-weighted spin-echo sequence with an echo time (TE) of 60 ms and pulse repetition time (TR) of 3000 ms. (d) MR images with a T₁-weighted spin-echo sequence with TE of 12 ms and TR of 300 ms.
Figure 12.14 (a) MR contrast images of MSC pellets (marked by the arrows) recorded on T₂-weighted sequences for MSCs. (b) T₁-weighted MR images of a rabbit before (left) and 30 min after (right) initial injection of a solution of ∼4 nm FeCo/GC NPs (metal dose ∼ 9.6 µmol kg⁻¹ for the ∼5-kg rabbit). The blood pool in the aorta is significantly brightened (positive contrast) in the MRI after injection. We also see signal increase in the kidney medulla and cortex due to the high blood volume within the kidney. We see little signal enhancement in the muscle.
Figure 12.15 (a) Graph of PAA-OA-coated Gd₂O₃ nanoparticles (2, 5, and 8 nm) showing high r₁ relaxivity values and the reference work for Gd-DTPA and PEG Gd₂O₃. (b) T₁-weighted MR images for oleic acid-coated 2-nm Gd₂O₃, PAA-OA-coated 2-nm Gd₂O₃, and Gd-DTPA (magnevist) depending on their Gd(III) concentration.
Figure 12.16 (a) T₁- and T₂-weighted imaging of glioma collected before and 1 h after intravenous injection of PEG-GdIO NPs with a dose of 5.0 mg kg⁻¹. (b) Contrast to noise analysis results.
Figure 12.17 Schematic structures of (a) Gd@C₈₂, (b) Gd@C₆₀, and (c) Gd₃N@C₈₀.

Chapter 13: Magnetic Iron Oxide Nanoparticles: Bioapplications and Potential Toxicity

Figure 13.1 (a) Synergistic enhancement of T₁ and T₂ contrast effects of GdIO nanoparticles under T₁- and T₂-weighted MR image; (b, c) T₁- and T₂-weighted in vivo MR images of BALB/c mice (top: coronal plane, bottom: transverse plane) before and after intravenous injection of GdIO nanoparticles with a dose of 2.0 mg kg⁻¹; (d, e) T₁- and T₂-weighted in vivo MR images of nude mice orthotopically inoculated with HepG2 liver cancer cells (sagittal plane) before and after intravenous injection of GdIO nanoparticles with a dose of 2.0 mg Fe kg⁻¹.
Figure 13.2 (a) Schematic representation of Doxorubicin (Dox) and Fe₃O₄ nanoparticle-loaded PEG-PLA micelles, with cRGD peptide conjugated on the micelle surface. (b) TEM image of cRGD-DOXO-SPIO-loaded polymeric micelles (scale bar: 20 nm). (c, d) Confocal laser scanning microscopy of SLK cells treated with 0% and 16% cRGD-DOXO-SPIO micelles (scale bars: 20 µm).
Figure 13.3 (a) TEM image of alkyl-PEI/SPIO nanocomposites (polymer/SPIO mass ratio = 0.6); (b) electron microscopic of cells labeled with SPIO probes (arrows), scale bar = 100 nm; (c) T₂-weighted gradient echo image shows a prominent hypointense area of labeled injection in the right frank (19 days after transplantation); (d) ratios of signal intensities of the control to the labeled injection.
Figure 13.4 Mechanism of cytotoxicity induced by SPIO nanoparticles.
Figure 13.5 Illustration depicting the assembly of polymers onto the surface of magnetic nanoparticle cores.
Figure 13.6 (a) Schematic illustration of the β-CD-Dex-g-SA/SPIO nanocomposite cross-sectional view; (b) TEM bright field image of SPIO micelles dried on a formvar coated copper grid (scale bar = 100 nm); (c) T₂-weighted MRI images (1.5 T, spin-echo sequence: TR = 5000 ms, TE = 12 ms) of β-CD-Dex-g-SA/SPIO micelles in water.

Chapter 14: Nanostructured Hydrogels for Diabetic Management

Figure Scheme 14.1 Typical mechanisms of glucose-sensitive hydrogels using GOD (a), Con A (b), and PBA (c and d) as glucose-sensing moieties.
Figure Scheme 14.2 Different approaches for application of glucose-sensitive LBL films in self-regulated insulin release.
Figure Scheme 14.3 Structure and sensing principle of the PCCA sensors designed by Asher et al. (a) and Zhang et al. (b).
Figure 14.1 (a) Synthesis of inverse opal hydrogel silica colloidal crystal as a template. (b, c) Reflection spectra (b) and photographs (c) of the gel in aqueous solution with different concentrations of glucose.
Figure 14.2 Shift of Fabry–Perot Fringes as a result of the analyte-induced swelling of the hydrogel film.
Figure 14.3 (a) Concept of bioartificial pancreas: encapsulation of islets semipermeable membrane for immunoisolation. (b) Islets enclosed in agarose microcapsules.
Figure 14.4 Encapsulation of pancreatic islets by LBL assembly of PLL-g-PEG and alginate.

Chapter 15: Inorganic Nanomaterials for Bone Tissue Engineering

Figure 15.1 SEM images of HAP samples with different shapes obtained at different pH value: (a) nanorods, pH = 7.0; (b) burr-like microspheres, pH = 5.0; (c) microflowers, pH = 4.5; (d) microsheets, pH = 4.0 [19].
Figure 15.2 SEM images of crystals grown on c-face substrate at 60 °C without and with aspartic acid. Nanostructures grown for 24 h without aspartic acid (a), for 24 h with 8.3 mmol dm⁻³ (b), for 24 h with 16.7 mmol dm⁻³ (c), and for 168 h with 8.3 mmol dm⁻³ (d) [59].
Figure 15.3 Fabrication of nano-HAP crystals on the surface of CaP scaffolds with designed morphologies via hydrothermal treatment in 0.2 M NaH₂PO₄ (S1), 0.2 M Na₃PO₄ (S2), and 0.2 M CaCl₂ (S3) aqueous solution. S0 as control without treatment [5].
Figure 15.4 (a) SEM image of MBG scaffolds and the interconnected large pores (300–500 µm) and (b) the well-ordered mesoporous structure (∼5 nm) in TEM.

Chapter 16: Nanotechnology in Coronary Artery Stent Coating

Figure 16.1 Ti–O film with n-type semiconductor characteristic has an electron-rich conduction band and low hole density of valence band, and hence prevent electrons transformed from fibrinogen, which possess narrow band characteristic.
Figure 16.2 (a) Design concept of HELIOS stent and (b) SEM images of Ti–O film-coated surface.
Figure 16.3 SEM images and schematic diagram of nanoporous and nanotube structure formulated by electrochemical self-ordering.
Figure 16.4 FITC-loaded NP-eluting stent [62].
Figure 16.5 Immunofluorescence staining of EPC, EC, and SMC surface antigens after 3 days of culture on material surface with different nanoparticle binding level, 316L SS was used as blank control. Dotted box shows the growth profile of three types of cells, which were selectively directed in a certain range of nanoparticle binding density.
Figure 16.6 The process of vascular healing. (a) Vascular injury or tissue ischemia leads to platelet adhesion and aggregation and subsequent activation of surrounding ECs. (b) Growth factors, chemokines, and proinflammatory factors secreted from platelets and activated ECs. (c) Concentration gradient of VEGF, SDF-1, and oxygen is formed between bone marrow and vascular niche. (d) VEGF and SDF-1 interacting with bone marrow cells cause the upregulation expression of proteases, including MMP-2, MMP-9, cathepsin G, and elastase. These proteases cleave cell–cell junctions and contribute to EPC isolate from bone marrow and mobilized into circulation. (e) Under the inducement of VEGF, SDF-1, and other inducing factors, EPCs mobilized into circulation, polarized, and migrated along with the cytokines concentration gradient. (f) EPCs migrated into vascular niche and homed to the injury site, where they adhered to the exposed basement membrane and participate in injury regeneration.

List of Tables

Chapter 1: Pharmacokinetics and Pharmacodynamics (PK/PD) of Bionanomaterials

Table 1.1 Drug transport process equtions [109]
Table 1.2 Category and feature of capillary wall from different organs [110]
Table 1.3 PK data of drug-loaded natural NMs
Table 1.4 PK data of drug-loaded synthetic NMs
Table 1.5 PK data of some NMs
Table 1.6 PK variations of the encapsulated drug among different NMs
Table 1.7 Variations between blood and tissue PK of NMs or the encapsulated drugs
Table 1.8 PK variations of the encapsulated drugs in the drug-NM systems
Table 1.9 PK variations between encapsulated drugs and other components in the drug-NM systems
Table 1.10 PK variations of NMs (or encapsulated drugs) via different routes of administration

Chapter 3: Polymeric Micelles for Drug Delivery

Table 3.1 Typical examples of polymeric micelles

Chapter 6: Antibody–Drug Conjugates

Table 6.1 Information about antibodies approved by FDA for cancer
Table 6.2 Structures of pH-responsive linkers in ADCs
Table 6.3 Structures of redox-responsive linkers in ADCs
Table 6.4 Structures of enzyme-responsive linkers in ADCs
Table 6.5 Structures of noncleavable linkers in ADCs
Table 6.6 Antigens targeted by ADCs in R&D process and their indication

Chapter 10: Nanographene in Biomedical Applications

Table 10.1 Drug and gene delivery based on functionalized nanographene

Chapter 12: Magnetic Nanostructures for MRI-Based Cancer Detection

Table 12.1 Characteristics of USPIO and SPIO agents: commercial or under clinical investigation [74]
Table 12.2 List of Gd(III)-based commercially available contrast agents

Chapter 16: Nanotechnology in Coronary Artery Stent Coating

Table 16.1 Novel surface coating used in vascular stent system
Table 16.2 Nanoparticle-based drug delivery in stent system

Edited by Yuliang Zhao and Youqing Shen

Biomedical Nanomaterials

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