Gold nanoparticles (AuNPs) exhibit localized surface plasmon resonance (LSPR) and are promising photothermal agents. However, the aggregation susceptibility and poor colloidal stability limit their biomedical applications, particularly in photothermal therapy. Here, a one-pot green method is employed to synthesize AuNPs using aqueous extracts of Coelastrella thermophila (CT) and Arthrospira sp. (SP) as reducing and stabilizing agents. These biogenic AuNPs (CT@AuNPs and SP@AuNPs), and their counterpart synthesized using a conventional procedure (Chem@AuNPs) have quasispherical shapes with hydrodynamic diameters of 26.8 +/- 10.3, 19.2 +/- 6.1, and 11.4 +/- 3.3 nm, respectively, and negatively charged surfaces. Fourier transform infrared spectroscopy reveals that CT@AuNPs and SP@AuNPs feature surface functional groups derived from algal proteins, carbohydrates, and fatty acids, which are associated with their greater colloidal stability (48 h) than Chem@AuNPs. These AuNPs elevate solution temperature up to similar to 8 degrees C under continuous green laser irradiation for 1 h. Cytotoxicity of CT@AuNPs and SP@AuNPs was lower toward Vero cells than toward HeLa cells, and under our experimental conditions, they induced cell death in similar to 70% of the latter upon green laser irradiation. Analyses using the 2 ',7 '-dichlorodihydrofluorescein diacetate and morphology suggest that the AuNP-induced cell death is attributed to LSPR-mediated hyperthermia and reactive oxygen species. Our alternative route to stable AuNPs addresses the persistent challenges posed by conventionally synthesized AuNPs in photothermal therapy and other fields.

Atherosclerosis remains the primary cause underlying cardiovascular diseases, however, there is no plaque-targeting pharmacotherapy clinically available. Nebivolol (NB), a third-generation beta-blocker clinically used to treat hypertension and heart failure, exerts potent antioxidative activities by inhibiting reactive oxygen species (ROS) production and scavenging ROS. Here, we developed an atherosclerotic plaque lesional macrophage-targeting nanotheranostic system that is coated with polydopamine (PDA) as a contrast agent to facilitate non-invasive photoacoustic imaging (PAI) of atherosclerotic plaques and NB as a drug to eliminate plaque ROS. In brief, mesoporous silica nanoparticles (MSNs) with interpenetrating chiral channels were coated with bioinspired PDA polymers (SPDA). PDA on the nanoparticle surface was then conjugated with hyaluronic acid (HA) to allow active targeting to atherosclerotic macrophages that overexpress CD44 (SPDA@HA). Upon loading of NB, the resulting NB/SPDA@HA nanoparticles could eliminate ROS and alleviate inflammation in activated macrophages. In addition, NB/SPDA@HA nanoparticles inhibited cell apoptosis and oxidized low density lipoprotein induced foam cell formation. Moreover, the silanols on the silica surface and the HA coating on the MSNs enable accelerated drug release from NB/SPDA@HA nanoparticles in response to the acidic and hyaluronidase-rich microenvironment in the plaque. Taken together, the dual-responsive NB/SPDA@HA nanotheranostic platform represents a promising nanomedicine for targeted atherosclerosis imaging and therapy.

Atherosclerosis is the primary cause of cardiovascular events such as heart attacks and strokes. However, current medical practice lacks non-invasive, reliable approaches for both imaging atherosclerotic plaques and delivering therapeutic agents directly therein. Here, a biocompatible and biodegradable pH-responsive nanoscale coordination polymers (NCPs) based theranostic system is reported for managing atherosclerosis. NCPs are synthesized with a pH-responsive benzoic-imine (BI) linker and Gd3+. Simvastatin (ST), a statin not used for lowering blood cholesterol but known for its anti-inflammatory and antioxidant effects in mice, is chosen as the model drug. By incorporating ST into the hydrophobic domain of a lipid bilayer shell on NCPs surfaces, ST/NCP-PEG nanoparticles are created that are designed for dual purposes: they diagnose and treat atherosclerosis. When administered intravenously, they target atherosclerotic plaques, breaking down in the mild acidic microenvironment of the plaque to release ST, which reduces inflammation and oxidative stress, and Gd-complexes for MR imaging of the plaques. ST/NCP-PEG nanoparticles show efficacy in slowing the progression of atherosclerosis in live models and allow for simultaneous in vivo monitoring without observed toxicity in major organs. This positions ST/NCP-PEG nanoparticles as a promising strategy for the spontaneous diagnosis and treatment of atherosclerosis. A dual-function nanoscale coordination polymers-based theranostic system, ST/NCP-PEG, is synthesized for managing atherosclerosis. This system can efficiently co-deliver gadolinium and simvastatin (ST) to the plaque and release both cargos therein in response to the acidic microenvironment, enabling spontaneous magnetic resonance imaging and efficacious treatment of atherosclerosis. Moreover, this system is biocompatible, biodegradable, and does not exert obvious toxicity. image

Introduction:Over 75% of clinical microbiological infections are caused by bacterial biofilms that grow on wounds or implantable medical devices. This work describes the development of a new poly(diallyldimethylammonium chloride) (PDADMAC)/alginate-coated gold nanorod (GNR/Alg/PDADMAC) that effectively disintegrates the biofilms of Staphylococcus aureus (S. aureus), a prominent pathogen responsible for hospital-acquired infections.Methods:GNR was synthesised via seed-mediated growth method, and the resulting nanoparticles were coated first with Alg and then PDADMAC. FTIR, zeta potential, transmission electron microscopy, and UV-Vis spectrophotometry analysis were performed to characterise the nanoparticles. The efficacy and speed of the non-coated GNR and GNR/Alg/PDADMAC in disintegrating S. aureus-preformed biofilms, as well as their in vitro biocompatibility (L929 murine fibroblast) were then studied.Results:The synthesised GNR/Alg/PDADMAC (mean length: 55.71 ± 1.15 nm, mean width: 23.70 ± 1.13 nm, aspect ratio: 2.35) was biocompatible and potent in eradicating preformed biofilms of methicillin-resistant (MRSA) and methicillin-susceptible S. aureus (MSSA) when compared to triclosan, an antiseptic used for disinfecting S. aureus colonisation on abiotic surfaces in the hospital. The minimum biofilm eradication concentrations of GNR/Alg/PDADMAC (MBEC50 for MRSA biofilm = 0.029 nM; MBEC50 for MSSA biofilm = 0.032 nM) were significantly lower than those of triclosan (MBEC50 for MRSA biofilm = 10,784 nM; MBEC50 for MRSA biofilm 5967 nM). Moreover, GNR/Alg/PDADMAC was effective in eradicating 50% of MRSA and MSSA biofilms within 17 min when used at a low concentration (0.15 nM), similar to triclosan at a much higher concentration (50 µM). Disintegration of MRSA and MSSA biofilms was confirmed by field emission scanning electron microscopy and confocal laser scanning microscopy.Conclusion:These findings support the potential application of GNR/Alg/PDADMAC as an alternative agent to conventional antiseptics and antibiotics for the eradication of medically important MRSA and MSSA biofilms.

Abstract Background Simvastatin is commonly used for the treatment of atherosclerosis. However, it causes side effects in some patients and not always effective. Moreover, although its treatment effects are often attributed to its cholesterol-lowering activity, some in vitro studies suggest that simvastatin also exerts antioxidant and anti-inflammatory effects. Purpose To develop an atherosclerotic plaque microenvironment-responsive nanoparticular system to deliver and release Gadolinium (Gd3+) for magnetic resonance imaging (MRI) and simvastatin for treatment within the plaque. Methods A nanoparticle was synthesized with nanoscale coordination polymers (NCPs), a pH-responsive linker, Gd3+, and simvastatin (ST), i.e. a ST/NCP-PEG nanoparticle. The biological effects of ST/NCP-PEG nanoparticle were tested on RAW264.7 macrophages. Four weeks old ApoE-/- mice were fed on high fat diet for 4 weeks and then randomly divided into three groups for a 8-week treatment regime via weekly intravenous administration of: 1) NCP-PEG nanoparticle without ST (control); 2) ST/NCP-PEG nanoparticles; or 3) free ST. Results We have developed a pH-responsive ST/NCP-PEG nanoparticle carrying an MRI contrast agent Gd3+ and a drug molecule ST, with an 80 nm diameter in spherical shape. This nanoparticle collapsed into biodegradable components while releasing both Gd3+ and ST under the atherosclerotic plaque microenvironment with a pH 5.6. ST/NCP-PEG nanoparticles exerted strong anti-oxidant and anti-inflammatory effects in vitro, as determined by an intracellular ROS indicator DCF-DA and expression levels of cytokines (e.g. IL-6, TNF-α and MCP-1), respectively. Upon intravenous administration, ST/NCP-PEG nanoparticles, as demonstrated by IVIS imaging system, accumulated specifically in the plaques in a time-dependent manner, reaching the peak concentration at 24 hours post administration. The plaques were readily visualized by MR imaging (T1-weighted signal) due to the enrichment of Gd3+. After 8 weeks treatment, compared to the empty NCP-PEG nanoparticle, treatment with ST/NCP-PEG and free ST reduced plaque lesion size by 55% (p<0.001) and 29% (p<0.05), respectively. Moreover, ST/NCP-PEG nanoparticles showed a larger reduction in plaque lesions than free ST (p<0.05). Oil red O staining of both the whole aorta and the aortic roots generated similar results in terms of plaque lesion changes. Mechanistically, both ST/NCP-PEG and free ST reduced ROS production and increased the ratios of M2/M1 macrophages within the plaque, with ST/NCP-PEG nanoparticle treatment exerted larger therapeutic effects. Moreover, long-term treatment with ST/NCP-PEG nanoparticles did not show obvious toxicity in all major organs. Conclusions An atherosclerotic plaque microenvironment-responsive MR imaging functional nanoparticle for targeted delivery of ST to the plaque could diagnose atherosclerotic plaque and improve treatment efficacy of ST.

One of the major weaknesses of therapeutic peptides is their sensitivity to degradation by proteolytic enzymes in vivo. Gold nanoparticles (GNPs) are a good carrier for therapeutic peptides to improve their stability and cellular uptake in vitro and in vivo. We conjugated the anticancer KT2 peptide as an anticancer peptide model to PEGylated GNPs (GNPs-PEG) and investigated the peptide stability, cellular uptake and ability of the GNPs-KT2-PEG conjugates to induce MDA-MB-231 human breast cancer cell death. We found that 11 nm GNPs protected the conjugated KT2 peptide from trypsin proteolysis, keeping it stable up to 0.128% trypsin, which is higher than the serum trypsin concentration (range 0.0000285 ± 0.0000125%) reported by Lake-Bakaar, G. et al., 1979. GNPs significantly enhanced the cellular uptake of KT2 peptides after conjugation. Free KT2 peptides pretreated with trypsin were not able to kill MDA-MB-231 cells due to proteolysis, while GNPs-KT2-PEG was still able to exert effective cancer cell killing after trypsin treatment at levels comparable to GNPs-KT2-PEG without enzyme pretreatment. The outcome of this study highlights the utility of conjugated anticancer peptides on nanoparticles to improve peptide stability and retain anticancer ability.

Studies have established that a serum protein corona pre-formed around gold nanorods (NRs) could be exploited for loading photosensitizers and chemotherapeutics to result in efficient cell kill in vitro with an extremely low dose. In this study, we further demonstrated that pre-forming a serum protein corona (PC) around citrate-capped NRs (NR-Cit) to form NR-PC conferred them stealth property and high hematocompatibility similar to the common strategy of PEGylating NRs, which would otherwise not be able to evade the immune system. Specifically, the NR-PC caused minimal complement activation with significantly lower formation of the terminal complement complex SC5b-9 measured in human serum containing NR-PC, and this resulted in low uptake by phagocytic U937 monocytes of 5.9% of the initial gold dose compared to 55.8% of NR-Cit. In addition, NR-PC exhibited very low hemolytic activity of less than 0.2% hemolysis with no observable effect on RBC morphology as opposed to 0.6% for NR-Cit at the same concentration of 1 nM NRs. Furthermore, we showed that the high hematocompatibility and stealth property of NR-PC were maintained even after the loading of small molecules, photosensitizer Chlorine e6 (Ce6), into the protein corona, thus further establishing the potential clinical relevance of exploiting the inevitably formed serum protein corona on nanoparticles as an effective delivery vector for small molecular therapeutics.

Understanding heat dissipation processes at nanoscale during cellular thermogenesis is essential to clarify the relationships between the heat and biological processes in cells and organisms. A key parameter determining the heat flux inside a cell is the local thermal conductivity, a factor poorly investigated both experimentally and theoretically. Here, using a nanoheater/nanothermometer hybrid made of a polydopamine encapsulating a fluorescent nanodiamond, we measured the intracellular thermal conductivities of HeLa and MCF-7 cells with a spatial resolution of about 200 nm. The mean values determined in these two cell lines are both 0.11 ± 0.04 W m-1 K-1, which is significantly smaller than that of water. Bayesian analysis of the data suggests there is a variation of the thermal conductivity within a cell. These results make the biological impact of transient temperature spikes in a cell much more feasible, and suggest that cells may use heat flux for short-distance thermal signaling.

The interaction between human serum albumin (HSA) and nanoparticles (NPs) to form HSA corona has widely been studied since endogenous functions of albumin are highly attractive for drug delivery. However, a full understanding of the molecular dynamics and factors behind the formation of HSA corona, including interactions between HSA and different surface ligands and between neighboring HSA molecules, resulting in conformational change of HSA is presently lacking. Here, we assembled 14 HSA molecules around gold nanorods (AuNRs) with different surface chemistries (bare gold surface, cetyltrimethylammonium bromide (CTAB), polystyrene sulfonate (PSS), and polydiallyldimethylammonium chloride (PDADMAC)) in silico and examined the dynamics of HSA corona formation using coarse-grained molecular dynamics for 300 ns of simulation. We observed that PDADMAC, being more flexible than PSS, resulted in all HSA molecules moving toward AuNR-PDADMAC, while the instability of CTAB on AuNR resulted in fewer HSA molecules moving toward AuNR-CTAB compared to AuNR-PSS. HSA molecules around AuNR-PDADMAC also exhibited the largest conformational change in terms of their radius of gyration (Rg) and root mean square deviation (RMSD). In the absence of surface ligands, HSA molecules around the bare AuNR were susceptible to steric hindrance with conformational change observed in terms of their RMSD but not their Rg unlike that of HSA molecules around AuNR-PDADMAC. The insights gained from the inclusion of neighboring HSA molecules in the simulation of corona formation could be more representative than examining a single adsorbed HSA molecule on AuNRs with different surface passivations.

Peptides and proteins have played an important role in many biological processes, functioning as enzymes, hormones, ligands, receptors, cell mediators, and structural components of cells. Being intrinsic molecules in signaling pathways, peptides allow for therapeutic intervention that closely mimic natural signaling cascades. However, the short chain of amino acids in free peptides is susceptible to proteolysis in vivo. Conjugation of peptides onto nanoparticles has been used as a strategy to extend peptide half-life through conferring steric hindrance and a high packing density that prevents proteolytic enzymes to degrade them. Here, we describe a method to conjugate the anticancer p53 peptides as our model peptide onto 12 nm gold nanoparticles (AuNPs) to form the AuNP-p53 peptide conjugate. Conjugation of the p53 short-chain peptide of 25 amino acids occurs through a combination of electrostatic interactions and covalent bonds between cysteine residues at the N-terminal of the peptide and the surface of the AuNPs. The AuNPs and AuNP-p53 are characterized by UV-Vis spectroscopy for its optical absorbance and zetasizer for their hydrodynamic diameter and zeta potential. The semiquantitative analysis of the amount of conjugated peptides on the AuNPs and peptide stability under trypsin treatment is performed on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

Understanding heat dissipation processes at the nanoscale during cellular thermogenesis requires the development of thermal sensing and heat releasing tools with nanometric resolution. Various approaches have been developed for nanoscale thermal sensing, among which fluorescent nanodiamond (FND) has emerged as a novel type of nanothermometer due to its biological compatibility, high sensitivity, extreme photostability and physicochemical inertness. On the other hand, the choice of methods that are able to release heat at nanoscale is somewhat limited. Usually, the released heat needs to be monitored on-site by adopting a separate approach to precisely measure the temperature. Here, by encapsulating fluorescent nanodiamond (FND) with a photothermal polymer, nanoheater and nanothermometer are integrated into a single nanoparticle. With the adoption of this dual-functionalized FND nanoparticle and optically detected magnetic resonance microscopy, repeatable heating while measuring the temperature with the accuracy approximately 1 degree Celsius in different environments, including intracellular milieu, is achieved in order to investigate their thermal conductivities. Our approach will contribute to clarifying the physical background how the heat released in a cell propagates through the intracellular space.

Serum albumin could potentially be exploited to form a protein corona on gold nanorods (AuNRs) for drug delivery because of its endogenous functionality as a small molecule carrier. However, the cetyltrimethylammonium bromide (CTAB) surfactant, which is a synthesis byproduct passivating AuNRs to confer colloidal stability, could also cause its conformational change upon interaction with serum albumin during the process of corona formation, thus altering its biological functions. Unfortunately, a clear understanding of how exactly human serum albumin (HSA) would change its conformation as it interacts with AuNR-CTAB is presently lacking. Here, we made use of coarse-grain molecular dynamics (CGMD) simulation to elucidate the interaction between HSA and AuNR-CTAB leading to its widely reported conformational change. We showed that HSA could sequester CTAB from the surface of AuNRs and form HSA-CTAB complexes, which could also interact with other adjacent complexes through "cross-linking" by the clusters of CTAB. Such a HSA-CTAB complex resulted in the observed conformational change of HSA, which we verified empirically with an esterase activity assay and by analyzing the root-mean-square-deviation of the HSA molecules from CGMD. The conformational change of HSA was not observed in AuNRs passivated with other negatively or positively charged surface ligands such as polystyrene sulfonate and polydiallyldimethylammonium chloride. Therefore, our study revealed that the conformational change experienced by HSA may not necessarily be attributed to protein unfolding on the surface of the AuNR due to charge interactions but rather to the instability of the surface ligands on the AuNRs which allows them to be sequestered by HSA to form HSA-CTAB complexes.

Charge and surface chemistry of gold nanorods (AuNRs) are often considered the predictive factors for cell membrane damage. Unfortunately, extensive research on AuNR passivated with polyelectrolyte (PE) ligand shell (AuNR-PE) has hitherto left a vital knowledge gap between the mechanical stability of the ligand shell and the cytotoxicity of AuNR-PEs. Here, the agreement between unbiased coarse-grained molecular dynamics (CGMD) simulation and empirical outcomes on hemolysis of red blood cells by AuNR-PEs demonstrates for the first time, a direct impact of the mechanical stability of the PE shell passivating the AuNRs on the lipid membrane rupture. Such mechanical stability is ultimately modulated by the rigidity of the PE components. The CGMD simulation results also reveal the mechanism where the PE chain adsorbs near the surface of the lipid bilayer without penetrating the hydrophobic core of the bilayer, which allows the hydrophobic AuNR core to be in direct contact with the hydrophobic interior of the lipid bilayer, thereby perforating the lipid membrane to induce membrane damage.

Aim:To establish a light-independent functionality of gold nanorods (AuNRs) with a human serum (HS) protein corona loaded with photosensitizer Chlorin e6 (AuNR-HS-Ce6) in M1 polarization of macrophages.Methods:RT-qPCR and ELISA were used to determine gene and protein expression, respectively. Uptake of AuNR-HS-Ce6 was determined via flow cytometry, inductively coupled plasma mass spectrometry and fluorescence microscopy. Cell viability was determined using PrestoBlue(R)cell viability assay.Results:An increase in M1 gene and protein expression was observed in AuNR-HS-Ce6-treated macrophages. Delivery of high Ce6 payload via AuNR-HS-Ce6 was the primary contributor toward M1 polarization. Finally, DLD-1 cells treated with conditioned media from AuNR-HS-Ce6-treated macrophages showed significantly reduced proliferation.Conclusion:Our study suggests an immunomodulatory potential of Ce6 in inducing light-independent M1 polarization outside of its role as a photosensitizer.

The emergence and rapid spread of antibiotic resistance poses a serious threat to healthcare systems across the globe. The existence of carbapenemase-producing Enterobacteriaceae (CPE) such as Klebsiella pneumoniae renders the use of carbapenems, the last-resort class of β-lactam antibiotics, ineffective against bacterial infections, often leading to CPE-associated mortalities. Current methods of detection such as the Carba NP test and modified Hodge's test require hours to days to detect, which delays the response to isolate patients for rapid intervention. Here, we developed a surface-enhanced Raman scattering (SERS)-based detection scheme which utilizes gold nanostars conjugated to a β-lactam antibiotic ceftriaxone (CRO) as a beacon for rapid detection of bacterial β-lactamase secreted by Delhi metalloproteinase (NDM)-producing Escherichia coli as our CPE model with carbapenemase activity. The cleavage of β-lactam ring in CRO by NDM (Class B β-lactamase) caused a detectable reduction in SERS intensities at 722, 1358, and 1495 cm-1 within 25 min. Ratiometric analysis of the SERS peaks at 722, 1358, and 1495 cm-1 normalized against the Raman peak of polystyrene cuvette at 620 cm-1 showed the peak at 1358 cm-1 having the most significant change in intensity upon CPE detection. This reduced detection time has not been reported to date for CPE detection, and our novel approach using SERS could be extended to detect the activity of other classes of β-lactamases to broaden its clinical utility.

Nanoparticle-based phototherapy has evolved to include immunotherapy as an effective treatment combination for cancers through inducing anti-cancer immune activation leading to downstream adaptive responses and immune protection. However, most cancer phototherapy studies that claimed anti-cancer immunogenic effects often included exogenous immunostimulants to potentiate immune responses and did not clearly establish their effects on immune cells. In this study, we showed that combined photodynamic (PDT) and photothermal therapy (PTT) using gold nanorods (NRs) loaded with the photosensitizer chlorin e6 (Ce6) on endogenously formed mouse serum (MS) protein coronas (i.e., NR-MS-Ce6) on EMT6 murine mammary carcinoma cells could potentiate the activation of both J774A.1 macrophages and DC2.4 dendritic cells. The activation of these innate immune cells by the conditioned media from cancer cells treated with combined PDT + PTT was cell-type and number dependent. While treated B16-OVA murine melanoma cells induced lower activation levels for both immune cell types compared to EMT6, they caused higher pro-inflammatory cytokine secretion levels. Our study suggests the importance of immunological investigations to complement any nanoparticle-based therapeutic interventions to better evaluate their efficacy. This could be achieved through a simple approach to screen for the first line of immune responses arising from these therapies prior to in vivo studies.

Mucus is an endogenous viscoelastic biopolymer barrier that limits the entry of foreign pathogens and therapeutic carriers to the underlying mucosal cells. This could be overcome with a hydrophilic and nonpositively charged carrier surface that minimizes interactions with the mucin glycoprotein fibers. Although PEGylation remains an attractive surface strategy to enhance mucopenetration, cell uptake of PEGylated nanoparticles (NPs) often remains poor. Here, we demonstrated polydopamine (PDA) coating to enhance both mucopenetration and cell uptake of NPs. PDA was polymerized on carboxylated polystyrene (PS) NPs to form a PDA coating, and the resulting PS-PDA achieved a similar level of mucopenetration as our PEGylated PS (PS-PEG) positive control in three separate studies: NP-mucin interaction test, transwell assay, and multiple particle tracking. Compared to water, the diffusions of PS-PDA and PS-PEG in reconstituted mucus solution were only 3.5 and 2.4 times slower, respectively, whereas the diffusion of bare PS was slowed by up to 250 times. However, the uptake of PS-PDA (61.2 ± 6.1%) was almost three times higher than PS-PEG (24.6 ± 5.4%) in T24 cells, which were used as a model for underlying mucosal cells. Our results showed a novel unreported functionality of PDA coating in enhancing both mucopenetration and cell uptake of NPs for mucosal drug delivery applications, not possible with conventional PEGylation strategies.

Urine voiding and the presence of a mucus layer on the apical surface of the urothelium are two major challenges towards an effective intravesical drug delivery for bladder malignancies. Improved bioavailability to the underlying bladder tissue could be achieved with delivery vectors that diffuse efficiently through the bladder mucus. Pegylation of delivery vectors remains the existing "gold standard" to enhance mucosal delivery despite known poor cell uptake and reported PEG sensitivity. Here, we showed improved mucopenetration of carboxylated polystyrene (PS) nanoparticles (NPs) passivated with a polydopamine (PDA) surface, at similar level as PEG. While the diffusion of PS NPs in mucus was retarded by ~1000-fold, PS-PDA diffused only 6-fold slower in mucus than water. This enabled faster and deeper penetration of PS-PDA into porcine bladder tissue beneath the mucus layer. The same PDA surface also conferred biocompatibility and enabled photothermal therapy (PTT) with significant surface disruption on an ex vivo porcine bladder model upon localized laser irradiation, which was not possible with PEG. Our outcomes suggested the facile and versatile PDA surface passivation of nanoparticles as an enabler for dual purposes of enhancing mucopenetration and allowing photothermal therapy on bladder tissue, which has not been demonstrated to date.

Therapeutic peptides suffer from major drawbacks such as peptide degradation in vivo due to proteolysis. Gold nanoparticles (AuNPs) are an effective carrier for therapeutic peptides that improve their stability in vivo, while also enabling nonspecific adsorption of complementary proteins to enhance their effectiveness. Using p53 peptide as a model known to disrupt the intracellular MDM2-p53 protein-protein interaction which tags the endogenous p53 proteins for degradation, we conjugated p53 peptides to AuNPs (AuNP-p53) and examined the functionality of AuNP-p53 to release the endogenous p53 proteins from being tagged for degradation, thereby increasing the level of stable p53 proteins in acute myeloid leukemia 2 (AML2) cells. We found that AuNPs did not just protect conjugated p53 peptides from trypsin degradation, but also helped to recruit 56.5% and 26.4% of total MDM2 and p53 proteins in the cells to form a protein corona around AuNP-p53. The proximity of MDM2/p53 complexes and p53 peptide on the surface of AuNP-p53 facilitated the action of p53 peptides to cause a sustained elevation of the p53 level in AML2 cells up to 6 h, which was not possible with free p53 peptide alone at the same concentration. Even a 20-fold higher concentration of free p53 peptide caused only a short-lived elevated p53 level of 1 h. The outcome of this study highlights the utility of combining conjugated ligands and complementary protein adsorption on nanoparticles to improve the biological functionality of the therapeutic ligands.