In Vitro Therapeutic Attributes of Luminescent Hydroxyapatite Nanoparticles in Codelivery Module.

Imminent prospects of clinical importance have been accomplished through divergent treatment modalities implemented using nanoscale platforms. In the present study, hydroxyapatite nanoparticles doped with copper nanoclusters (HAPs) were explored for codelivery of a hydrophobic drug, namely, norfloxacin (NX), and a hydrophilic photosensitizer, such as methylene blue (MB). NX and MB were successfully homed into HAPs (MB-NX-HAPs), which further exhibited a pH-dependent release of both. With the objective of attaining an enhanced effect, MB-NX-HAPs were evaluated for combination therapy, involving chemotherapy and photodynamic therapy (PDT) with irradiation at 640 nm. The combinatorial therapy approach was initially applied for antibacterial therapy, which suggested a considerable reduction in bacterial growth of Gram-negative strain Pseudomonas aeruginosa MTCC 2488. Thereafter, the antiproliferative study performed in cancer cell lines (HeLa and MCF-7) revealed the efficiency of MB-NX-HAPs in bestowing a combinatorial effect through chemotherapy and PDT (irradiation at 640 nm). The combined effect exerted through MB-NX-HAPs subsequently induced reactive oxygen species (ROS) generation, cell cycle alteration, and apoptosis activation in cancer cells. The biocompatible nature of MB-NX-HAPs was appreciably shown through their minimal effect on the normal cell line (HEK-293). Additionally, HAPs through luminescence of copper nanoclusters were suggested to aid in bioimaging of cancer cell lines.

Quercetin-Loaded Luminescent Hydroxyapatite Nanoparticles for Theranostic Application in Monolayer and Spheroid Cultures of Cervical Cancer Cell Line In Vitro.

Nanoscale materials have been explored as better alternatives to conventional therapeutic agents in cancer theranostics in the recent period due to efficacy in overcoming biological, biomedical, and biophysical barriers. Analysis on the ability of copper nanocluster (CuNC)-doped hydroxyapatite nanoparticles (Cu-HXNPs) as suitable nanocarriers for anticell proliferative application was carried out. Having high adsorption capacity, the Cu-HXNPs could be loaded with the anticancer drug quercetin, which is a polyphenolic flavonoid compound, and were used as nanocarriers to be applied on HeLa (cancer cells) and HEK-293 (normal cells). The drug release profile was found to be pH-dependent, where maximum release of quercetin from quercetin-loaded Cu-HXNPs was observed in acidic pH as compared to physiological pH. The Cu-HXNPs could release quercetin, which could effectively decline proliferation of cancer cells via generation of reactive oxygen species. Moreover, the released quercetin significantly altered the cell cycle pattern and triggered the cells to undergo apoptosis. Additionally, the efficacy of Cu-HXNPs as a nanocarrier to release quercetin on 3D spheroids of HeLa had been checked, which demonstrated significant reduction in the viability of 3D spheroids. The luminescent CuNCs used for doping HXNPs endowed the nanocarrier with the imaging property, which was an excellent feature in confirming their uptake by the cells. Thus, the study suggested Cu-HXNPs to be a beneficial nanocarrier for both bioimaging and therapeutic purpose in the field of cancer theranostics.

Magnetotactic T-Budbots to Kill-n-Clean Biofilms.

Treatment of persistent biofilm infections has turned out to be a formidable challenge even with broad-spectrum antibiotic therapies. In this direction, intelligent micromachines may serve as active mechanical means to dislodge such deleterious bacterial communities. Herein, we have designed biocompatible micromotors from tea buds, namely, T-Budbots, which shows the capacity to be magnetically driven on a biofilm matrix and remove or fragment biofilms with precision, as a part of the proposed non-invasive "Kill-n-Clean" strategy. In a way, we present a bactericidal robotic platform decorated with magnetite nanoparticles aimed at clearing in vitro biofilms present on the surfaces. We have also shown that the smart porous T-Budbots can integrate antibiotic ciprofloxacin due to electrostatic interaction on their surface to increase their antibacterial efficacy against dreadful pathogenic bacterial communities of Pseudomonas aeruginosa and Staphylococcus aureus. It is noteworthy that the release of this drug can be controlled by tuning the surrounding pH of the T-Budbots. For example, while the acidic environment of the biofilm facilitates the release of antibiotics from the porous T-Budbots, the drug release was rather minimal at higher pH. The work represents a first step in the involvement of a plant-based microbot exhibiting magneto-robotic therapeutic properties, providing a non-invasive and safe approach to dismantle harmful biofilm infections.

Smartphone controlled interactive portable device for theranostics in vitro.

In this work, a smartphone controlled interactive theranostic device has been developed to perform in vitro photodynamic therapy (PDT) and diagnostic assays for treatment assessment on a single platform. Further, silver nanorod (Ag NR) was identified as a photosensitizer and its effect was studied in three different cell lines. PDT was achieved with Ag NRs using low irradiation (1.4 mW/cm2 at 632 nm) from light emitting diodes (LEDs) in the device. Specifically, PDT in conjugation with widely used chemotherapeutic drug doxorubicin (Dox) proved effective in killing of HeLa cancer cells and multicellular tumor spheroids at a minimum dose of Ag (2.5 µg/mL). The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and LDH (lactate dehydrogenase) assays performed with the device indicated the therapeutic success of the delivered PDT. The device is portable and can be adapted for different wavelength irradiations and radiation doses. Additionally, wireless operation using a custom designed smartphone application makes it convenient to use in complex environments without much of human intervention.

Copper Nanocluster-Doped Luminescent Hydroxyapatite Nanoparticles for Antibacterial and Antibiofilm Applications.

Novel strategies in the field of nanotechnology for the development of suitable multifunctional drug delivery vehicles have been pursued with promising upshots. Luminescent copper nanocluster-doped hydroxyapatite nanoparticles (HAP NPs) were synthesized and applied for the delivery of antibacterial drug kanamycin. The negatively charged doped HAP NPs could electrostatically interact with the positively charged kanamycin. The kanamycin-loaded doped HAP NPs showed pronounced activity in the case of Gram-negative bacteria compared to that in Gram-positive bacteria. Upon interaction with the bacteria, kanamycin could probably generate harmful agents such as hydroxyl radical that leads to bacterial cell damage. After being incorporated with copper nanoclusters (Cu NCs), the doped HAP NPs were applied for the bioimaging of bacterial cells. The biocompatibility of doped HAP NPs was also studied in HeLa cells. As compared to copper nanoclusters, the doped HAP NPs showed excellent biocompatibility even at higher concentrations of copper. The kanamycin-loaded doped HAP NPs were further applied toward Pseudomonas aeruginosa biofilm eradication. Thus, the as-synthesized copper nanocluster-doped HAP NPs were applied as nanocarriers for antibiotic drug delivery, bioimaging, and antibiofilm applications.

Gold-Nanocluster-Embedded Mucin Nanoparticles for Photodynamic Therapy and Bioimaging.

Effective delivery of a photosensitizer with the ability to trace its eventual progress forms an important aspect in photodynamic therapy (PDT). Further, the delivery mechanism might require possessing the ability to traverse through the complex mucus barrier that offers retention of therapeutic molecules. In this work, gold nanocluster (Au NC)-embedded mucin nanoparticles were synthesized by a rapid green synthetic procedure for application as nanocarriers and to achieve image-guided PDT. The mucin-based nanocarrier exhibited excellent biocompatibility toward normal cells (HEK 293T). The photosensitizer methylene blue (MB) was loaded onto these Au NC-mucin nanoparticles (NPs). HeLa cancer cells were treated with MB-loaded Au NC-mucin nanoparticles under irradiation of 640 nm light. The cell viability assay revealed that the viability of HeLa cells was reduced to 50% after treatment with MB-loaded Au NC-mucin NPs under 640 nm irradiation. The luminescence exhibited by Au NCs in the nanocarrier was applied for tracking the delivery of MB inside the HeLa cells using confocal microscopy. The flow cytometry assays elucidated the mechanism of cell death.