Multifunctional Photonic Nanomaterials for Diagnostic, Therapeutic, and Theranostic Applications.

The last decade has seen dramatic progress in the principle, design, and fabrication of photonic nanomaterials with various optical properties and functionalities. Light-emitting and light-responsive nanomaterials, such as semiconductor quantum dots, plasmonic metal nanoparticles, organic carbon, and polymeric nanomaterials, offer promising approaches to low-cost and effective diagnostic, therapeutic, and theranostic applications. Reasonable endeavors have begun to translate some of the promising photonic nanomaterials to the clinic. Here, current research on the state-of-the-art and emerging photonic nanomaterials for diverse biomedical applications is reviewed, and the remaining challenges and future perspectives are discussed.

Dual-Color-Emitting Carbon Nanodots for Multicolor Bioimaging and Optogenetic Control of Ion Channels.

The development of intrinsically multicolor-emitting carbon nanodots (CNDs) has been one of the great challenges for their various fields of applications. Here, the controlled electronic structure engineering of CNDs is performed to emit two distinct colors via the facile surface modification with 4-octyloxyaniline. The so-called dual-color-emitting CNDs (DC-CNDs) can be stably encapsulated within poly(styrene-co-maleic anhydride) (PSMA). The prepared water-soluble DC-CNDs@PSMA can be successfully applied to in vitro and in vivo dual-color bioimaging and optogenetics. In vivo optical imaging can visualize the biodistribution of intravenously injected DC-CNDs@PSMA. In addition, the light-triggered activation of ion channel, channelrhodopsin-2, for optogenetic applications is demonstrated. As a new type of fluorophore, DC-CNDs offer a big insight into the design of charge-transfer complexes for various optical and biomedical applications.

Hyaluronate-Peanut Agglutinin Conjugates for Target-Specific Bioimaging of Colon Cancer.

Colon cancer is one of the most common death-related cancers in the world. For treating colon cancer, it is crucial to detect and remove malignant lesions early. Here, we developed hyaluronate (HA)-peanut agglutinin (PNA) conjugates for the bioimaging of colon cancer. The HA-PNA conjugates were successfully synthesized by the coupling reaction between aldehyde-modified HA and the N-terminal amine group of PNA. For diagnostic imaging, rhodamine B (RhoB) was chemically conjugated onto PNA in HA-PNA conjugates. After intraluminal injection of HA-PNA-RhoB conjugates into tumor-bearing mice, small-sized colon cancers could be effectively visualized by ex vivo imaging with an in vivo imaging system (IVIS) and a two-photon microscope. With these results taken together, we could confirm the feasibility of HA-PNA-RhoB conjugates as a bioimaging agent for detecting colon cancers.

Hyaluronate and its derivatives for customized biomedical applications.

Since hyaluronate (HA) was firstly isolated from the vitreous of bovine eyes in 1934, HA has been widely investigated for various biomedical applications. As a naturally-occurring polysaccharide, HA has been used for joint lubrication and ocular treatment in its intact form due to the excellent biocompatibility, viscoelasticity, biodegradability, and hygroscopic properties. HA can be easily functionalized via the chemical modification of its carboxyl and hydroxyl groups. Recently, a variety of biological functions of HA have been explored and a number of customized applications have been investigated taking advantages of the interaction between HA and biological tissues. HA has been used for drug delivery to enhance the blood circulation time of drugs with target-specificity to HA receptors in the body. HA has been also used to prepare tissue engineering hydrogel scaffolds for the spatiotemporal control of encapsulated cells. In this review, we describe the key biological functions of HA in the body in terms of its structure, physical properties, biodistribution and interaction with HA receptors. After that, we describe unique advantages that allow HA to be applied in various biomedical fields. Finally, we report the conventional and newly emerging applications of HA and its derivatives under commercial development stages.

Targeted Hyaluronate-Hollow Gold Nanosphere Conjugate for Anti-Obesity Photothermal Lipolysis.

Obesity is associated with the risk of developing several severe diseases, such as metabolic disorder, diabetes, and heart diseases. Despite wide investigation and trials, a noninvasive obesity therapy is still an important medical unmet need, targeting the abnormal adipose tissue. Here, we developed hyaluronate-hollow gold nanosphere-adipocyte-targeting peptide (HA-HAuNS-ATP) conjugates for the photothermal ablation of adipose tissues. The HA-HAuNS-ATP conjugate could be noninvasively delivered into the skin and effectively target to adipocytes in the subcutaneous. With near-infrared laser illumination, HA-HAuNS-ATP conjugate enabled highly effective photothermal ablation of adipose tissues in C57BL/6 obesity mice. The photoacoustic imaging confirmed the successful transdermal delivery and the photothermal lipolysis of HA-HAuNS-ATP conjugate. Taken together, the transdermal HA-HAuNS-ATP conjugate might have a great potential for noninvasive photothermal lipolysis.

Biodegradable Nitrogen-Doped Carbon Nanodots for Non-Invasive Photoacoustic Imaging and Photothermal Therapy.

Multifunctional nanoparticles have been widely investigated for biomedical applications, such as imaging, therapy, and drug delivery. Especially, photoactive nanoparticles have received great attention as theranostic agents because of their heat-generating abilities after exposure to laser irradiation. However, photostability and safety issues have been the technical hurdles for further clinical applications. Here, we designed nitrogen (N)-doped carbon nanodots (N-CNDs) that have strong absorption in the near-infrared region, high photostability, and excellent biodegradability. Optimized N-CNDs can be utilized not only as a new photoacoustic (PA) imaging agent but also as a superior photothermal therapy (PTT) agent in vivo because of their strong optical absorption at a specific wavelength. We used N-CNDs to perform in vivo/ex vivo noninvasive PA imaging of sentinel lymph nodes via local delivery and performed PTT for cancer ablation therapy. Finally, biodegradation and renal clearance were confirmed by performing whole-body PA monitoring and a degradation test.

Two-photon microscopy of a Flt1 peptide-hyaluronate conjugate.

AIM: Two-photon microscopy was performed to visualize ocular distribution of Flt1 peptide-hyaluronate (HA) conjugate micelles for eye drop treatment of corneal neovascularization. MATERIALS & METHODS: Flt1 peptide-HA conjugate micelles were topically administered to the eye for two-photon microscopy and antiangiogenic effect assessment after silver nitrate cauterization. RESULTS: In vivo two-photon microscopy revealed that Flt1 peptide-HA conjugate micelles were absorbed and remained on the corneal epithelia with an increased residence time, facilitating the corneal delivery of carboxyfluorescein succinimidyl ester (CFSE) as a model drug. Furthermore, repeated eye drops of Flt1 peptide-HA conjugate micelles showed comparable therapeutic effect to the subconjunctival injection on the corneal neovascularization. DISCUSSION & CONCLUSION: We confirmed the feasibility of Flt1 peptide-HA conjugate micelles for eye drop treatment of corneal neovascularization.

Photodynamic therapy of melanoma skin cancer using carbon dot - chlorin e6 - hyaluronate conjugate.

Despite wide application of photodynamic therapy (PDT) for the treatment of melanoma skin cancers, there are strong biomedical unmet needs for the effective generation of singlet oxygen after targeted delivery of photosensitizers. Here, we investigated a facile PDT of melanoma skin cancer using transdermal carbon dot - chlorine e6 - hyaluronate (Cdot-Ce6-HA) conjugates. The Cdot-Ce6-HA conjugate was synthesized by the coupling reaction of diaminohexane modified HA (DAH-HA) with the carboxylic group of Ce6. The singlet oxygen generation of Cdot-Ce6-HA conjugates in aqueous solution was more significant than that of free Ce6. The enhanced transdermal and intracellular delivery of Cdot-Ce6-HA conjugates to B16F10 melanoma cells in tumor model mice were corroborated by confocal microscopy and two-photon microscopy. The laser irradiation after topical treatment with Cdot-Ce6-HA conjugates resulted in complete suppression of melanoma skin cancers. The antitumor effect was confirmed by histological analysis with H&E staining and TUNEL assay for tumor apoptosis. Taken together, we could confirm the feasibility of Cdot-Ce6-HA conjugate for transdermal PDT of melanoma skin cancers. STATEMENT OF SIGNIFICANCE: To our knowledge, this is the first report on a facile transdermal photodynamic therapy (PDT) of melanoma skin cancer using carbon dot - chlorine e6 - hyaluronate (Cdot-Ce6-HA) conjugates. We found that the singlet oxygen generation of Cdot-Ce6-HA conjugates in aqueous solution was more significant than that of free Ce6. Confocal microscopy and two-photon microscopy clearly confirmed the enhanced transdermal and intracellular delivery of Cdot-Ce6-HA conjugates to B16F10 melanoma cells in tumor model mice. Taken together, we could confirm the feasibility of Cdot-Ce6-HA conjugate for transdermal PDT of melanoma skin cancers.

Hyaluronic acid-gold nanoparticle/interferon α complex for targeted treatment of hepatitis C virus infection.

Gold nanoparticles (AuNPs) have been extensively investigated as an emerging delivery carrier of various biopharmaceuticals. Instead of nonspecific polyethylene glycol (PEG) conjugated interferon α (IFNα) for the clinical treatment of hepatitis C virus (HCV) infection, in this work, a target-specific long-acting delivery system of IFNα was successfully developed using the hybrid materials of AuNP and hyaluronic acid (HA). The HA-AuNP/IFNα complex was prepared by chemical binding of thiolated HA and physical binding of IFNα to AuNP. According to antiproliferation tests in Daudi cells, the HA-AuNP/IFNα complex showed a comparable biological activity to PEG-Intron with a highly enhanced stability in human serum. Even 7 days postinjection, HA-AuNP/IFNα complex was target-specifically delivered and remained in the murine liver tissue, whereas IFNα and PEG-Intron were not detected in the liver. Accordingly, HA-AuNP/IFNα complex significantly enhanced the expression of 2',5'-oligoadenylate synthetase 1 (OAS1) for innate immune responses to viral infection in the liver tissue, which was much higher than those by IFNα, PEG-Intron, and AuNP/IFNα complex. Taken together, the target-specific HA-AuNP/IFNα complex was thought to be successfully applied to the systemic treatment of HCV infection.

Bioimaging of hyaluronic acid derivatives using nanosized carbon dots.

Fluorescent nanosized carbon dots (Cdots) are an emerging bioimaging agent with excellent chemical inertness and marginal cytotoxicity in comparison to widely used semiconductor quantum dots. In this work, we report the application of Cdots for real time bioimaging of target specific delivery of hyaluronic acid (HA) derivatives. Polyethylene glycol (PEG) diamine-capped Cdots were synthesized by the pyrolysis of citric acid in a hot solvent. The synthesized Cdots showed strong fluorescence under UV excitation with emission properties dependending on the excitation wavelength. HA-Cdot conjugates were synthesized by amide bond formation between amine groups of Cdot and carboxylic groups of HA. After confirmation of the negligible cytotoxicity of Cdots and HA-Cdot conjugates, in vitro bioimaging was carried out for target specific intracellular delivery of the HA-Cdot conjugates by HA receptor-mediated endocytosis. Furthermore, in vivo real-time bioimaging of Cdots and HA-Cdot conjugates exhibited the target specific delivery of HA-Cdot conjugates to the liver with abundant HA receptors. Taken together, we could confirm the feasibility of HA derivatives as a target-specific drug delivery carrier for the treatment of liver diseases and Cdots as a promising bioimaging agent.

In vivo real-time confocal microscopy for target-specific delivery of hyaluronic acid-quantum dot conjugates.

Hyaluronic acid (HA), which is a biocompatible, biodegradable, and linear polysaccharide in the body, has been widely used for various biomedical applications. In this work, real-time bioimaging for target-specific delivery of HA derivatives was carried out using quantum dots (QDs). In vitro confocal microscopy of HA-QD conjugates confirmed the intracellular delivery of HA derivatives to B16F1 cells with HA receptors by HA-receptor-mediated endocytosis. Furthermore in vivo real-time confocal microscopy of HA-QD conjugates successfully visualized the target specific delivery and accumulation of HA-QD conjugates from the fluorescence-labeled blood vessels to the liver tissues. The authors could confirm the feasibility of HA derivatives as a target-specific intracellular drug-delivery carrier for the treatment of liver diseases and the in vivo real-time confocal microscopy as a new bioimaging tool for various drug-delivery applications. FROM THE CLINICAL EDITOR: This study demonstrates the possibility of labeling hyaluronic acid with quantum dots for visualization and for targeted intracellular drug delivery in liver disease models.