Advances in photoacoustic imaging aided by nano contrast agents: special focus on role of lymphatic system imaging for cancer theranostics.

Photoacoustic imaging (PAI) is a successful clinical imaging platform for management of cancer and other health conditions that has seen significant progress in the past decade. However, clinical translation of PAI based methods are still under scrutiny as the imaging quality and clinical information derived from PA images are not on par with other imaging methods. Hence, to improve PAI, exogenous contrast agents, in the form of nanomaterials, are being used to achieve better image with less side effects, lower accumulation, and improved target specificity. Nanomedicine has become inevitable in cancer management, as it contributes at every stage from diagnosis to therapy, surgery, and even in the postoperative care and surveillance for recurrence. Nanocontrast agents for PAI have been developed and are being explored for early and improved cancer diagnosis. The systemic stability and target specificity of the nanomaterials to render its theranostic property depends on various influencing factors such as the administration route and physico-chemical responsiveness. The recent focus in PAI is on targeting the lymphatic system and nodes for cancer diagnosis, as they play a vital role in cancer progression and metastasis. This review aims to discuss the clinical advancements of PAI using nanoparticles as exogenous contrast agents for cancer theranostics with emphasis on PAI of lymphatic system for diagnosis, cancer progression, metastasis, PAI guided tumor resection, and finally PAI guided drug delivery.

The Role of Ultrasound in Cancer and Cancer-Related Pain-A Bibliometric Analysis and Future Perspectives.

Ultrasound has a deep penetrating ability with minimal or no tissue injury, while cancer-mediated complications during diagnosis, therapy, and surgery have become a serious challenge for clinicians and lead to the severity of the primary condition (cancer). The current study highlights the importance of ultrasound imaging and focused ultrasound therapy during cancer diagnosis, pain reduction, guidance for surgical resection of cancer, and the effectiveness of chemotherapy. We performed the bibliometric analysis on research domains involving ultrasound, cancer management, pain, and other challenges (chemotherapy, surgical guidance, and postoperative care), to observe the trend by which the research field has grown over the years and propose a possible future trend. The data was obtained from the Web of Science, processed, and exported as plain text files for analysis in the Bibliometrix R web interface using the Biblioshiny package. A total of 3248 documents were identified from 1100 journal sources. A total of 390 articles were published in 2022, with almost a 100% growth rate from previous years. Based on the various network analysis, we conclude that the outcome of the constant research in this domain will result in better patient care during the management of various diseases, including cancer and other co-morbidities.

Combined effect of polyacrylic acid and vitamin E in preventing calcium oxalate crystal deposition in the kidneys of experimental hyperoxaluric rats.

Our study explores the combined effect of polyacrylic acid and vitamin E as prophylactic and curative agent against ethylene glycol (EG) induced calcium oxalate stone formation in Wistar rats. Male Wistar rats (54) were divided into nine groups, including control. The experimental groups were equally segregated into two for preventive study (4) and curative study (4). Urolithiasis was induced with 0.75% (v/v) EG in drinking water. Polyacrylic acid (10 mg/kg) and vitamin E (300 IU/day) were supplemented from day 1 for preventive and day 30 for curative studies. Restoration of urinary lithogenic factors (calcium, oxalate, phosphate, citrate and magnesium) and renal function (urea and creatinine in serum) by intervening agents were accomplished compared to urolithic rats (P < .001). Abnormal localization and increased expression of Tamm-Horsfall Protein, osteopontin and transferrin were observed in the kidneys of urolithic rats (P < .001) from immunohistochemistry and immunoblotting analysis. Polyacrylic acid and vitamin E supplementation have regulated the expression of the urinary macromolecules. Pro-inflammatory cytokines in kidney were significantly reduced (P < .001) by the intervening agents compared to urolithic rats. Therefore, polyacrylic acid and vitamin E in combination could be a potential formulation for better management of urolithiasis.

Beneficial effect of Citrus limon peel aqueous methanol extract on experimentally induced urolithic rats.

CONTEXT: Citrus limon (L.) Burm.f. (Rutaceace) is a commonly available fruit variety with high medicinal and industrial values. OBJECTIVE: Lemon peel (LP) extract was studied as a potent preventive and curative agent for experimentally induced hyperoxaluric rats. MATERIALS AND METHODS: Gas chromatography-mass spectrometry (GC-MS) analyses and toxicity study were performed for aqueous methanol LP extract. Twenty-four Wistar rats were segregated into four groups. Group 1: Control; Group 2: Urolithic (ethylene glycol (EG) - 0.75%); Group 3: Preventive study (EG + LP extract administration from 0th to 7th week); Group 4: Curative study (EG + LP extract administration from 4th to 7th week). Animals received LP extract daily by oral administration (100 mg/kg body weight) for 7 weeks. RESULTS AND DISCUSSION: GC-MS analyses revealed that compound 6 was abundant in the LP extract (32%) followed by compound 1 (∼21%). The LD50 value of LP extract was found to be >5000 mg/kg of body weight. Urolithic rats showed significantly higher urinary calcium and oxalate (4.47 ± 0.44 and 18.86 ± 0.55 mg/24 h, respectively) excretion compared with control and experimental rats. Renal function parameters like urea (84 ± 8.5 and 96.1 ± 3.6 mg/dL), creatinine (1.92 ± 0.27 and 1.52 ± 0.22 mg/dL), and urinary protein (2.03 ± 0.02 and 2.13 ± 0.16 mg/24 h) were also reduced by LP extract (p < 0.001) and corroborated with tissue analyses (SOD, catalase, and MDA levels) and histological studies in normal and experimental animals. Immunohistochemical staining of THP and NF-κB in urolithic animals showed elevated expression than the control, while LP extract suppressed the expression of these proteins. CONCLUSION: In conclusion, lemon peel is effective in curing kidney stone disease and also can be used to prevent the disease and its recurrence.

Exosomes and ultrasound: The future of theranostic applications.

Biomaterials and pertaining formulations have been very successful in various diagnostic and therapeutic applications because of its ability to overcome pharmacological limitations. Some of them have gained significant focus in the recent decade for their theranostic properties. Exosomes can be grouped as biomaterials, since they consist of various biological micro/macromolecules and possess all the properties of a stable biomaterial with size in nano range. Significant research has gone into isolation and exploitation of exosomes as potential theranostic agent. However, the limitations in terms of yield, efficacy, and target specificity are continuously being addressed. On the other hand, several nano/microformulations are responsive to physical or chemical alterations and were successfully stimulated by tweaking the physical characteristics of the surrounding environment they are in. Some of them are termed as photodynamic, sonodynamic or thermodynamic therapeutic systems. In this regard, ultrasound and acoustic systems were extensively studied for its ability towards altering the properties of the systems to which they were applied on. In this review, we have detailed about the diagnostic and therapeutic applications of exosomes and ultrasound separately, consisting of their conventional applications, drawbacks, and developments for addressing the challenges. The information were categorized into various sections that provide complete overview of the isolation strategies and theranostic applications of exosomes in various diseases. Then the ultrasound-based disease diagnosis and therapy were elaborated, with special interest towards the use of ultrasound in enhancing the efficacy of nanomedicines and nanodrug delivery systems, Finally, we discussed about the ability of ultrasound in enhancing the diagnostic and therapeutic properties of exosomes, which could be the future of theranostics.

Ketogenic Diet: A Promising Neuroprotective Composition for Managing Alzheimer's Diseases and its Pathological Mechanisms.

Ketogenic diet and ketone bodies gained significant attention in recent years due to their ability to influence the specific energy metabolism and restoration of mitochondrial homeostasis that can help in hindering the progression of many metabolic diseases, including diabetes and neurodegenerative diseases. A ketogenic diet consists of high fat and low carbohydrate contents, which makes the body glucose deprived and rely on alternative sources (ketone bodies) for energy. It has been initially designed and supplemented for the treatment of epilepsy, and, later, its influence on many energyderiving biochemical pathways made it a highly sorted food supplement for many metabolic diseases and even for bodybuilding and calorie restriction in healthy individuals. Among the reported therapeutic action over a range of diseases, neurodegenerative disorders, especially Alzheimer's disease, gained the attention of many researchers and clinicians because of the higher benefits of the ketogenic diet on this disease. Complex pathology and multiple influencing factors of Alzheimer's disease make exploration of its therapeutic strategies a demanding task. It was a common phenomenon that energy deprivation in neurological disorders, including Alzheimer's disease, progress rapidly. The ability of ketone bodies to stabilize the mitochondrial energy metabolism makes it a suitable intervening agent. In this review, we will discuss various research progress made with regards to ketone bodies/ketogenic diet for the management of Alzheimer's disease and elaborate in detail about the mechanisms that are influenced during their therapeutic action.

Andrographolide Relieves Post-Operative Wound Pain but Affects Local Angiogenesis.

Andrographolide (Andro), the major constituent of Andrographis paniculata Nees (Acanthaceae), is was known to reduces inflammatory reaction. In the current study, the ability of Andro to reduce pain sensation in a rat post-operative wound model was explored. The hind paws of 18 Sprague-Dawley rats (SD) bearing post-operative wounds received the following three treatments: Saline, Andro via direct injection into the paw (Andro-injected) and Tablet containing Andro + poly (lactic-co-glycolic acid) (PLGA) (Andro-tablet). Von Frey tests assessed mechanical allodynia at 1, 3, 5 h and 1-, 2-, 3-, 4-, and 5-days post-operation. Behavioral analyses were performed to measure reaction threshold and reaction frequencies. Immunoreactivity of p-ERK and GluR1 was examined in the dorsal horn of the spinal cord. Histopathological and immunostaining studies were conducted on paw epidermis to observe the gross morphology and angiogenesis. The threshold for inducing allodynia increased and the reaction frequency reduced in the Andro-injected group compared to the saline-group, at 3 h post-surgery and the effect lasted between 3-4 days. The threshold for inducing pain and reaction frequency for the Andro-tablet group did not differ from the saline-treated group. The levels of p-ERK and GluR1 in the dorsal horn were reduced after Andro treatment. No significant difference in wound healing index was observed between saline and Andro-injected groups, but CD-31 staining showed less angiogenesis in the Andro-injected group. Andro significantly reduced mechanical allodynia compared to saline treatment, both in shorter and longer time frames. Furthermore, Andro influenced the expression of p-ERK and GluR1 in the dorsal horn, and the angiogenesis process in the wound healing area.

nCP:Fe Nanocontrast Agent for Magnetic Resonance Imaging-Based Early Detection of Liver Cirrhosis and Hepatocellular Carcinoma.

Early detection of liver tumors and cirrhotic lesions by magnetic resonance imaging (MRI) remains a great challenge. Here, we report a biomineral nanocontrast agent based on iron-doped nanocalcium phosphate (nCP:Fe-CA) for magnetic resonance imaging of early-stage liver cirrhotic and hepatocellular carcinoma nodules using rat models. We have optimized an intravenously injectable, aqueous suspension of nCP:Fe-CA having an average size of 137.6 nm, a spherical shape, magnetic relaxivity of 63 mM-1S-1, and colloidal stability for 48 h, post-resuspension in an aqueous phase. Compared to superparamagnetic iron oxide nanoparticles (SPIONs), the optimized nCP:Fe-CA could detect liver tumor lesions as small as ∼0.25 cm, whereas the current clinical detection limit is ∼1 cm. In addition, multiple cirrhotic nodules of size <0.2 cm could be detected by nCP:Fe-CA-assisted MRI. The number of nodules observed after injecting nCP:Fe-CA was ∼3 times higher than that without CA (5-10 nodules). A biocompatibility study on healthy rats injected with nCP:Fe-CA showed unaltered liver transaminases, blood urea nitrogen, serum creatinine, and insignificant hemolysis. Furthermore, hepatobiliary clearance of nCP:Fe-CA was observed in 72 h compared to prolonged retention of SPIONs for 30 days when tested under identical conditions. Overall, the nCP:Fe-CA nanoparticles showed promising results as a biocompatible, MR contrast (T2) agent for the early-stage imaging of liver cirrhosis and hepatocellular carcinoma.

Effects of Water Extract of Cynanchum paniculatum (Bge.) Kitag. on Different Breast Cancer Cell Lines.

Cynanchum paniculatum (Bge.) Kitag. (CP) is an important medicinal herb used in Chinese herbal medicine, with a variety of biological activities including anticancer property. In this study, we explored the water extract of CP, for its anticancer effects against breast cancer cells with different mutation types. Cells were grouped as untreated (Control); CP direct treatment (dir-CP); Conditioned medium from CP treated (sup-CP), and untreated cells (sup-Control). Effects of dir-CP and sup-CP were compared to corresponding untreated cells on cytotoxicity, cell migration, and protein expression (cleaved caspase-3, caspase-9, and MMP-2 and 9). CP treatment showed time-dependent decrease in cell number of MDA-MB-231 and SK-Br-3 (both ER(-) PR(-)), while the decrease in cell number was not as significant in MCF-7 and ZR-75-1 cells (both ER(+) PR(+)). sup-CP treatment inhibited the cell migration of MDA-MB-231 and MCF-7 (Her2(-)) in a 24 h scratch assay. Our data suggested that ER(-) PR(-) cells are more sensitive to the CP in terms of direct cytotoxicity, which is not regulated by caspase-3. CP inhibited the migration of the two Her2(-) cells, and this correlated with MMP-2 regulation. The migration of ER(-) PR(-) cells was more sensitive to conditioned medium with CP treatment than to direct CP, and this is not regulated by MMP-2. Our data suggested that CP has anticancer potential on various breast cancer cells through different mechanisms and is specifically effective in inhibiting the migration of the triple negative MDA-MB-231. Our data provide insight into the mechanism of CP against breast cancer progression and would benefit the medical practitioners in better management with CP usage.

Kaempferitrin-Treated HepG2 Differentially Expressed Exosomal Markers and Affect Extracellular Vesicle Sizes in the Secretome.

Kaempferitrin is extracted in significantly high quantities from the leaves of Cinnamomum osmophloeum, which belongs to a group of plant species that comes under the genus Cinnamomum, well-known for its established anti-diabetic property in Chinese medicine. Oral administration of kaempferitrin and Cinnamomum osmophloeum extract reduced blood sugar in alloxan-induced diabetic rats and improved the lipid profile in hamsters respectively. In this paper we studied the differential protein expression profile using mass spectrometry approach in the kaempferitrin-treated conditioned medium of liver cancer cell line HepG2. We discovered that 33 genes were up/down-regulated consistently between two biological samples. A slightly different version of the analysis software selected 28 genes, and the final 18 genes that appeared in both lists were selected. Interestingly, 5 proteins out of 18 were either exosomal markers or reported in high frequency of occurrence in exosome/secreted vesicles. We also examined the extracellular particles with atomic force microscopy (AFM), which showed that the conditioned medium of kaempferitrin treated had larger vesicles and fewer small vesicles. Expression of some lipid-regulating genes were also altered. Our data suggested that extracellular vesicle secretions may be regulated by kaempferitrin, and regulation of lipid profile by kampeferitrin involves multiple mechanisms.

Evaluation of garlic oil in nano-emulsified form: Optimization and its efficacy in high-fat diet induced dyslipidemia in Wistar rats.

Garlic oil nanoemulsion was formulated using ultrasonic emulsification and the optimized garlic oil nanoemulsion ratio (1:2) of oil: surfactant showed spherical, with tiny droplet size 24.9 ± 1.11 nm. It was observed that the prepared nanoemulsion has the zeta potential of -42.63 ± 1.58 mV and a low polydispersity index of 0.2 ± 0.09 with excellent stability. The formulation was subjected to in vivo acute and sub-acute toxicity. In acute toxicity study, single oral administration of 18.63 ml of garlic oil nanoemulsion/kg resulted in immediate mortality. However, garlic oil nanoemulsion (0.46 ml/kg) and tween 80 (0.5 ml/kg) administered rats did not exhibit any toxicity and showed no changes in hematological and histological parameters. Further, both preventive and curative studies of garlic oil nanoemulsion were evaluated in high-fat diet fed dyslipidemic Wistar rats. Garlic oil nanoemulsion administered groups showed a significant effect in reducing the levels of lipid profiles (p < 0.001) compared to atorvastatin and garlic oil. Evaluation of lipid deposits in hepatic tissues was analyzed by Oil Red O staining, which revealed that garlic oil nanoemulsion administered rats markedly reduced the fat depots. Our findings suggest that garlic oil nano-emulsified form reduced toxicity and improved efficacy in preventing and treating dyslipidemia.

Biomineral Nano-Theranostic agent for Magnetic Resonance Image Guided, Augmented Radiofrequency Ablation of Liver Tumor.

Theranostic nanoparticles based on biocompatible mineral compositions can significantly improve the translational potential of image guided cancer nano-therapy. Here, we report development of a single-phase calcium phosphate biomineral nanoparticle (nCP) with dual-mode magnetic resonance contrast (T1-T2) together with radiofrequency (RF) mediated thermal response suitable for image-guided RF ablation of cancer. The nanoparticles (NP) are engineered to provide dual MR contrast by an optimized doping concentration (4.1 at%) of paramagnetic ion, Fe3+, which also renders lossy dielectric character for nCP leading to thermal response under RF exposure. In vivo compatibility and dual-mode MR contrast are demonstrated in healthy rat models. MRI and T2 mapping suggest hepatobiliary clearance by ~96 hours. MRI guided intratumoral injection in subcutaneous rat glioma and orthotopic liver tumor models provide clear visualization of NP in MRI which also helps in quantifying NP distribution within tumor. Furthermore, by utilising RF mediated thermal response, NP treated tumor could be ablated using clinically approved RF ablation system (10 W,13.3 GHz). Real-time in vivo thermal imaging exhibits 119 ± 10% increase in temperature change (ΔT) for NP treated orthotopic liver tumor (ΔT = 51.5 ± 2 °C), compared to untreated healthy liver control (ΔT = 21.5 ± 2 °C). In effect, we demonstrate a promising nano-biomineral theranostic agent for dual-mode MRI combined with radiofrequency ablation of solid tumors.

Polyacrylic acid attenuates ethylene glycol induced hyperoxaluric damage and prevents crystal aggregation in vitro and in vivo.

The study explores calcium oxalate crystal inhibiting characteristic of polyacrylic acid (pAA), an anionic polymer in in vitro and in vivo. Animals were divided into 5 groups where group 1 served as control, group 2 were made hyperoxaluric by supplementing with Ethylene glycol (EG) 0.75% (v/v) for 30 days. Group 3, 4 & 5 were also given with EG and treated simultaneously with 2.5, 5 & 10 mg of pAA/kg of body weight, respectively. Urine, serum and tissue analyses along with histological studies were performed at the end of the 30 days study. In vitro crystallization was significantly inhibited by pAA and further it was supported by particle size analyses, XRD and FT-IR studies. Toxicological analyses showed that pAA was safe to use in animals at concentrations below 100 mg/kg BW. In vivo anti-urolithic study showed significant improvement in urinary lithogenic factors (calcium, oxalate, phosphate, citrate & magnesium) and renal function parameters (creatinine, urea and protein). Tissue analyses on anti-oxidant enzyme activity and lipid peroxides showed maintenance of tissue antioxidant status in the pAA supplemented rats and histological studies demonstrated the nephroprotection offered by pAA and were concurrent to the biochemical analyses. Supplementation of pAA not only reduces the crystal aggregation but also regulates the expression and localization of crystal inhibiting proteins and gene expression of inflammatory cytokines in experimental animals. In summary, pAA is a potent anti-urolithic agent in rats and we can propose that 10 mg/kg body weight is the effective dosage of pAA and this concentration can be used for further studies.

Regulation of urinary crystal inhibiting proteins and inflammatory genes by lemon peel extract and formulated citrus bioflavonoids on ethylene glycol induced urolithic rats.

The objective of this study is to check the regulation of crystal matrix proteins and inflammatory mediators by citrus bioflavonoids (CB) and Lemon peel (LP) extract in hyperoxaluric rats. The animals were divided into six groups with 6 animals each. Group 1: Control, Group 2: Urolithic (Ethylene glycol (EG)-0.75%); Group 3 & 5: Preventive study (EG + CB (20 mg/kg body weight) and LP (100 mg/kg body weight) extract administration from 0th-7th week) respectively; Group 4 & 6: Curative study (EG + CB and LP extract administration from 4th-7th week) respectively by oral administration. Urinary lithogenic factors (Calcium, oxalate, phosphate and citrate) were normalized in CB & LP supplemented rats, while serum parameters revealed the nephroprotective nature of the intervening agents compared to urolithic rats (p < 0.001). Immunoblotting studies showed significantly increased expression of THP, osteopontin and transferrin in kidneys of urolithic rats (p < 0.001), while preventive and curative study showed near normal expression of these proteins. Expression of NF-κB, TNF-α and IL-6 were raised significantly (p < 0.001), while a very minimal increase in MCP-1 expression was observed in urolithic rats compared to control. Hence, supplementation of CB and LP reduced the crystal promoting factors and provides protection from crystal induced renal damage.