An Optimized Method for Microcomputed Tomography Analysis of Trabecular Parameters of Metal Scaffolds for Bone Ingrowth.

Owing to its superior mechanical and biological properties, titanium metal is widely used in dental implants, orthopedic devices, and bone regenerative materials. Advances in 3D printing technology have led to more and more metal-based scaffolds being used in orthopedic applications. Microcomputed tomography (µCT) is commonly applied to evaluate the newly formed bone tissues and scaffold integration in animal studies. However, the presence of metal artifacts dramatically hinders the accuracy of µCT analysis of new bone formation. To acquire reliable and accurate µCT results that reflect new bone formation in vivo, it is crucial to lessen the impact of metal artifacts. Herein, an optimized procedure for calibrating µCT parameters using histological data was developed. In this study, the porous titanium scaffolds were fabricated by powder bed fusion based on computer-aided design. These scaffolds were implanted in femur defects created in New Zealand rabbits. After 8 weeks, tissue samples were collected to assess new bone formation using µCT analysis. Resin-embedded tissue sections were then used for further histological analysis. A series of deartifact two-dimensional (2D) µCT images were obtained by setting the erosion radius and the dilation radius in the µCT analysis software (CTan) separately. To get the µCT results closer to the real value, the 2D µCT images and corresponding parameters were subsequently selected by matching the histological images in the particular region. After applying the optimized parameters, more accurate 3D images and more realistic statistical data were obtained. The results demonstrate that the newly established method of adjusting µCT parameters can effectively reduce the influence of metal artifacts on data analysis to some extent. For further validation, other metal materials should be analyzed using the process established in this study.

Iron Oxide Nanoparticles as Imaging and Therapeutic Agents for Atherosclerosis.

Atherosclerosis is the major cause of cardiovascular diseases and is the leading cause of mortality worldwide. Iron oxide nanoparticles have emerged as potential diagnostic and therapeutic agents for a wide range of conditions. To date, the theranostic applications of iron oxide nanoparticles have been studied mainly in cancer, but atherosclerosis has not received the same attention. Therefore, it appears appropriate to review the current and future applications of iron oxide nanoparticles for the diagnosis and therapy of atherosclerosis. This review will first discuss current imaging techniques for the diagnosis of atherosclerosis as well as their limitations. It will then discuss the role of nanotechnology for molecular imaging of atherosclerosis and the benefits of this approach as well as reviewing current developments in the field including single, bi-, and tri-modal imaging. Next, it will discuss the role of nanotechnology for therapies of atherosclerosis with a focus on nanotheranostics, concluding with a look at the challenges faced by nanoparticle-based imaging and therapy of atherosclerosis as well as a look at future prospects.

Gene expression profiling of tumor stroma interactions in retinoblastoma.

We aimed to identify the critical molecular pathways altered upon tumor stroma interactions in retinoblastoma (RB). In vitro 2 D cocultures of RB tumor cells (Weri-Rb-1 and NCC-RbC-51) with primary bone marrow stromal cells (BMSC) was established. Global gene expression patterns in coculture samples were assessed using Affymetrix Prime view human gene chip microarray and followed with bioinformatics analyses. Key upregulated genes from Weri-Rb-1 + BMSC and NCC-RbC-51 + BMSC coculture were validated using qRT-PCR to ascertain their role in RB progression. Whole genome microarray experiments identified significant (P ≤ 0.05, 1.1 log 2 FC) transcriptome level changes induced upon coculture of RB cells with BMSC. A total of 1155 genes were downregulated and 1083 upregulated in Weri-Rb-1 + BMSC coculture. Similarly, 1865 genes showed downregulation and 1644 genes were upregulation in NCC-RbC-51 + BMSC coculture. The upregulated genes were significantly associated with pathways of focal adhesion, PI3K-Akt signalling, ECM-receptor interaction, JAK-STAT, TGF-ß signalling thus contributing to RB progression. Validation of key genes by qRT-PCR revealed significant overexpression of IL8, IL6, MYC and SMAD3 in the case of Weri-Rb-1 + BMSC coculture and IL6 in the case of NCC-RbC-51 + BMSC coculture. The microarray expression study on in vitro RB coculture models revealed the pathways that could be involved in the progression of RB. The gene signature obtained in a stimulated model when a growing tumor interacts with its microenvironment may provide new horizons for potential targeted therapy in RB.

Evidence of Tumour Microenvironment and Stromal Cellular Components in Retinoblastoma.

BACKGROUND: The tumour microenvironment (TME) consisting of tumour cells and multiple stromal cell types regulate tumour growth, invasion and metastasis. While the concept of TME and presence of stromal cellular components is widely established in cancers, its significance in the paediatric intraocular malignancy, retinoblastoma (RB), remains unknown. METHODS: The study qualitatively identified the presence of multiple stromal cellular subtypes in RB TME by immunohistochemistry. RESULTS: Results of the study identified the presence of stromal cell types such as endothelial cells, tumour-associated macrophages, fibroblasts, cancer-associated fibroblasts, retinal astrocytes and glia in RB TME. The extent of stromal marker positivity, however, did not correlate with histopathological features of RB. CONCLUSIONS: The findings of the study convincingly suggest the presence of a stromal component in RB tumours. The interactions between stromal cells and tumour cells might be of profound importance in RB progression.

Synthesis of saporin-antibody conjugates for targeting EpCAM positive tumour cells.

Epithelial cell adhesion molecule (EpCAM) is a transmembrane glycoprotein involved in cell proliferation and differentiation. Ribosomal inactivating proteins derived from plants specifically target ribosomes and irreversibly inhibit protein synthesis. EpCAM antibody and saporin were conjugated using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/N-hydroxysuccinimide chemistry. The mass of the conjugates were characterised using matrix-assisted laser desorption ionisation (MALDI). The saporin-EpCAM (SAP-EpAB) conjugates were tested in-vitro against MCF-7 (breast cancer cells), WERI-Rb1 (retinoblastoma) cells. The flow cytometry and fluorescence microscopy were performed to show the binding efficiency of SAP-EpAB conjugate. Whole transcriptome changes of sap-conjugate treated cells were studied using affymetrix microarrays. MALDI-TOF analysis and polyacrylamide gel electrophoresis confirmed the conjugation of SAP with EpCAM antibody. Flow cytometry and fluorescent microscopy analysis revealed the binding of SAP-EpAB conjugates to the MCF-7, WERI-Rb1 cells. Apoptosis assay by annexin-V has shown an increased apoptotic and necrotic population in conjugate treated cells. MTT assay confirmed the tumour cell death and had shown the IC50 value of 0.8 µg for conjugate in MCF-7 (breast cancer cells), and 1 µg for WERI-Rb1 (retinoblastoma) cells. The microarray analysis revealed downregulation of the tumourigenic genes and upregulation of pro-apoptotic genes leading to apoptosis of tumour cells.

Utility of nanomedicine targeting scar-forming myofibroblasts to attenuate corneal scarring and haze.

Corneal scarring refers to the loss of normal corneal tissue, replaced by fibrotic tissue (during wound repair) thereby affecting corneal transparency and vision quality. The corneal wound healing process involves a complex series of physiological events resulting in the transformation of transparent keratocytes into opaque myofibroblasts; the prominent cause of irregular extracellular matrix synthesis leading to the development of corneal opacity/hazy vision. Globally, corneal scarring/haze is one of the most prevalent causes of blindness. Ocular trauma (physical and chemical) and microbial infections induce corneal tissue damage. Although great progress has been made in the clinical management of ocular diseases, the global rates of corneal blindness remain high, nonetheless. The topical conventional modalities treating corneal wounds/injuries have inherent limitations/side effects such as low bioavailability of a therapeutic agent, upregulation of the intraocular pressure and the toxicity/allergy of the drug. These limitations/side effects rather than treating the wound, often negatively affect the healing process, especially, when applied frequently for longer periods. Recently, there has been an increasing evidence provided by the preclinical studies that nanotechnology-based drug-delivery systems can improve drug bioavailability, through controlled drug release and targeted delivery. After reviewing the epidemiology, risk factors of corneal scarring/haze and the conventional ocular medicines, we review here the different nanodrug-delivery systems and potential drug candidates including nanoherbal formulations investigated for their efficacy to heal the damaged cornea. Finally, we discuss the challenges of using these nanomedicinal platforms.

Correction to: Engineered atherosclerosis-specific zinc ferrite nanocomplex-based MRI contrast agents.

After publication of the original article [1], the authors found that Figure 2e (Hsp70-Ch-Lf-ZF in Jurkat cells, 2 h), Figure 5b (HSP70-Lf-ZF) and Figure 5c (control) contained incorrect images. This does not affect the figure legends, results and conclusions of the article.

Novel ß-Ti35Zr28Nb alloy scaffolds manufactured using selective laser melting for bone implant applications.

Titanium (Ti) based tissue engineering scaffolds can be used to repair damaged bone. However, successful orthopedic applications of these scaffolds rely on their ability to mimic the mechanical properties of trabecular bone. Selective laser melting (SLM) was used to manufacture scaffolds of a new ß-Ti35Zr28Nb alloy for biomedical applications. Porosity values of the scaffolds were 83% for the FCCZ structure (face centered cubic unit cell with longitudinal struts) and 50% for the FBCCZ structure (face and body centered cubic unit cell with longitudinal struts). The scaffolds had an elastic modulus of ∼1 GPa and a plateau strength of 8-58 MPa, which fall within the values of trabecular bone (0.2-5 GPa for elastic modulus and 4-70 MPa for compressive strength). The SLM-manufactured ß-Ti35Zr28Nb alloy showed good corrosion properties. MTS assay revealed that both the FCCZ and FBCCZ scaffolds had a cell viability similar to the control. SEM observation indicated that the osteoblast-like cells adhered, spread and grew healthily on the surface of both scaffolds after culture for 7, 14 and 28 d, demonstrating good biocompatibility. Overall, the SLM-manufactured Ti35Zr28Nb scaffolds possess promising potential as hard-tissue implant materials due to their appropriate mechanical properties, good corrosion behavior and biocompatibility. STATEMENT OF SIGNIFICANCE: Novel ß Ti35Zr28Nb alloy scaffolds with FCCZ and FBCCZ structures were successfully fabricated by selective laser melting (SLM) for biomedical applications. The scaffolds showed values of elastic modulus of ∼1 GPa and plateau strength of 8-58 MPa, which fall within the ranges of the mechanical properties of trabecular bone. The SLM-manufactured ß Ti35Zr28Nb alloy showed good corrosion properties. Both SLM-manufactured FCCZ and FBCCZ scaffolds exhibited good biocompatibility, with osteoblast-like cells attaching, growing, and spreading in a healthy way on their surfaces after culturing for different periods up to 28 d.

Neocarzinostatin, Aptamer Conjugates for Targeting EpCAM-positive Tumor Cells.

BACKGROUND/AIM: The aim of this study was to investigate the role of Neocarzinostatin (NCS) conjugated with epithelial cell adhesion molecule (EpCAM) aptamer in EpCAM-positive cancer cells. NCS is an antitumor antibiotic protein chromophore that has the ability to cleave double stranded DNA and can be used as a potential drug for the treatment of EpCAM-positive cancers. EpCAM aptamer is an oligonucleotide ligand that binds specifically to EpCAM, a protein overexpressed in tumor cells. MATERIALS AND METHODS: NCS was conjugated with EpCAM aptamer using Sulfo-Succinimidyl 6-(3-(2-pyridyldithio) - propionamide hexanoate) LC-(SPDP) cross-linker to deliver it to EpCAM-positive tumor cells. The conjugates were characterized using polyacrylamide gel electrophoresis (PAGE) and high-performance liquid chromatography (HPLC). Flow cytometry was used to study the binding efficiency of the aptamer and the conjugates in cancer cells. The effect of the conjugate on cancer cells was studied using propidium iodide (PI) to analyze the cell cycle phase changes. The apoptosis assay was performed using the IC50 concentration of NCS. Microarrays were performed to study the gene level changes in cancer cells upon treatment with NCS and the conjugate. RESULTS: Flow cytometry revealed significant binding of aptamer and conjugate in the MCF-7 and WERI-Rb1 cell lines. Briefly, 62% in MCF and 30% in WERI-Rb1 cells with conjugate treated cells (p<0.005). The cell-cycle analysis indicated G2 phase arrest in MCF-7 cells and S phase arrest in WERI-Rb1 cells (p<0.005). Microarray analysis showed differentially expressed genes involved in cell cycle, DNA damage, and apoptosis. The BrDU assay and the apoptosis assay showed that the expression of BrDU was reduced in conjugate-treated cells and the PARP levels were increased confirming the double stranded DNA breaks (p<0.005). In MCF-7 and WERI-Rb1 cells, most of the cells underwent necrosis (p<0.005). CONCLUSION: The EpCAM aptamer conjugated NCS showed specificity to EpCAM-positive cells. The effect of the conjugates on cancer cells were impressive as the conjugate arrested the cell cycle and promoted apoptosis and necrosis. The high levels of PARP expression confirmed the DNA breaks upon conjugate treatment. Our study demonstrates that the NCS conjugated with EpCAM can be targeted to cancer cells sparing normal cells.

Targeted Theranostics Against Solid Cancer Using Metal Bond Milk Protein and Aptamers.

Targeted therapeutics is a new generation therapy that can increase the efficacy of chemotherapeutic drugs, by facilitating site-specific delivery with minimum off-target effects. Amongst the targeted therapy, nucleic-acid based aptamers are increasingly gaining interest due to their small size, long shelf-life and ease in synthesis. Further, lactoferrin, a milk protein belonging to the transferrin family is now an established multi-functional iron-binding protein. Its applicability as an immunomodulator, antimicrobial and an anti-cancer agent has made this protein highly valuable. Recent research has been focusing towards increasing the efficacy of lactoferrin by encapsulating them in novel nanoparticles, that facilities in providing their controlled and sustained release. This review focuses on the application of aptamers against solid tumors, specifically colorectal cancer (CRC) indicating the different anti-cancer targeted strategies to target anti-angiogenic vascular endothelial growth factor -A (VEGF-A) and epidermal growth factor receptor (EGFR) signalling. Additionally, it highlights the synergistic approach of functionalising aptamers with drug loaded nanoparticles to facilitate enhanced uptake, stability and increase in the retention time. Special emphasis is given on lactoferrin loaded aptamer functionalised nanoparticles as anticancer drug delivery systems. Apart from highlighting the role of these aptamernanocarriers in tumor specific targeting and induction of apoptosis, there applicability in nanotheranostics, involving detection, diagnosis and treatment is also discussed here.

In Vitro and In Vivo Assessment of Docetaxel Formulation Developed for Esophageal Stents.

Esophageal cancer (EC) mostly affects the elderly population and is frequently diagnosed at an advanced stage. Self-expanding metal stents (SEMS) are the most popular mode of palliation, but they are associated with reocclusion caused by tumor growth. To overcome this problem, docetaxel (DTX)-loaded polyurethane formulations were prepared for stent application. The films were evaluated against the cancer cell lines, OE-19 and OE-21, and normal esophageal cell line Het-1A. The DTX and the formulations were evaluated in vitro for the cytotoxicity and in vivo in nude mice. It was found that DTX and the formulations have a weak activity against the EC cell lines and an even weaker activity against Het-1A cell line. Preliminary in vivo studies showed skin toxicity in nude mice necessitating modification of the formulation. Reevaluation in a mouse xenograft model resulted in toxicity at high dose formulations while the low dose formulation exhibited modest advantage over commercial IV formulation; however, there was no significant difference between the commercial IV and blank formulation. DTX combination with an anti-cancer agent having complementary mode of action and non-overlapping toxicity could yield better outcome in future.

SurR9C84A protects and recovers human cardiomyocytes from hypoxia induced apoptosis.

Survivin, as an anti-apoptotic protein and a cell cycle regulator, is recently gaining importance for its regenerative potential in salvaging injured hypoxic cells of vital organs such as heart. Different strategies are being employed to upregulate survivin expression in dying hypoxic cardiomyocytes. We investigated the cardioprotective potential of a cell permeable survivin mutant protein SurR9C84A, for the management of hypoxia mediated cardiomyocyte apoptosis, in a novel and clinically relevant model employing primary human cardiomyocytes (HCM). The aim of this research work was to study the efficacy and mechanism of SurR9C84A facilitated cardioprotection and regeneration in hypoxic HCM. To mimic hypoxic microenvironment in vitro, well characterized HCM were treated with 100µm (48h) cobalt chloride to induce hypoxia. Hypoxia induced (HI) HCM were further treated with SurR9C84A (1µg/mL) in order to analyse its cardioprotective efficacy. Confocal microscopy showed rapid internalization of SurR9C84A and scanning electron microscopy revealed the reinstatement of cytoskeleton projections in HI HCM. SurR9C84A treatment increased cell viability, reduced cell death via, apoptosis (Annexin-V assay), and downregulated free cardiac troponin T and MMP-9 expression. SurR9C84A also upregulated the expression of proliferation markers (PCNA and Ki-67) and downregulated mitochondrial depolarization and ROS levels thereby, impeding cell death. Human Apoptosis Array further revealed that SurR9C84A downregulated expression of pro-apoptotic markers and augmented expression of HSPs and HTRA2/Omi. SurR9C84A treatment led to enhanced levels of survivin, VEGF, PI3K and pAkt. SurR9C84A proved non-toxic to normoxic HCM, as validated through unaltered cell proliferation and other marker levels. Its pre-treatment exhibited lesser susceptibility to hypoxia/damage. SurR9C84A holds a promising clinical potential for human cardiomyocyte survival and proliferation following hypoxic injury.

Protozoa lectins and their role in host-pathogen interactions.

Lectins are proteins/glycoproteins of non-immune origin that agglutinate red blood cells, lymphocytes, fibroblasts, etc., and bind reversibly to carbohydrates present on the apposing cells. They have at least two carbohydrate binding sites and their binding can be inhibited by one or more carbohydrates. Owing to carbohydrate binding specificity of lectins, they mediate cell-cell interactions and play role in protozoan adhesion and host cell cytotoxicity, thus are central to the pathogenic property of the parasite. Several parasitic protozoa possess lectins which mediate parasite adherence to host cells based on their carbohydrate specificities. These interactions could be exploited for development of novel therapeutics, targeting the adherence and thus helpful in eradicating wide spread of protozoan diseases. The current review highlights the present state knowledge with regard to protozoal lectins with an emphasis on their haemagglutination activity, carbohydrate specificity, characteristics and also their role in pathogenesis notably as adhesion molecules, thereby aiding the pathogen in disease establishment.

Amoebiasis vaccine development: A snapshot on E. histolytica with emphasis on perspectives of Gal/GalNAc lectin.

Amoebiasis/amebiasis is a gastrointestinal infection caused by an enteric dwelling protozoan, Entamoeba histolytica. The disease is endemic in the developing world and is transmitted mainly via the faecal-oral route (e.g., in water or food) and may or may not be symptomatic. This disease of socio-economic importance worldwide involves parasite adherence and cytolysis of human cells followed by invasion that is mediated by galactose-binding (Gal/GalNAc) surface lectin. Disruption of the mucus layer leads to invasive intestinal and extraintestinal infection. Gal-lectin based vaccinations have conferred protection in various animal models against E. histolytica infections. Keeping in view the pivotal role of Gal/GalNAc lectin in amoebiasis vaccine development, its regulation, genomic view of the parasite involving gene conversion in lectin gene families, current knowledge about involvement of Gal/GalNAc lectin in adherence, pathogenicity, signalling, encystment, generating host immune response, and in turn protozoa escape strategies, and finally its role as effective vaccine candidate has been described. This review will help researchers to explore pathogenesis mechanism along with genomic studies and will also provide a framework for future amoebiasis vaccine development studies.

Mushroom Lectins as Promising Anticancer Substances.

Lectins are proteins/glycoproteins of non-immune origin, which are widely distributed in nature. They have at least one non-catalytic domain, which binds reversibly to specific monosaccharides or oligosaccharides. Lectins recognizing sugar moieties in cell walls or cell membranes alter the membrane physiology and trigger biochemical changes in the cell. Thus, various applications of lectins have been described, for example as tools to identify aberrant glycans expressed by neoplastic cells and as antitumor agents by inducing apoptosis by various mechanisms. In order to widen applications of anti-tumor lectins, a detailed investigation of their action mechanism is required. Mushrooms are a valuable source of novel lectins with unique specificities and potentials for biotechnological and biomedical applications. This article reviews information on anti-proliferative activity of mushroom lectins obtained in-vitro and in-vivo. The possible role of lectins as cancer therapeutics is discussed together with the mechanisms underlying the anti-proliferative activity, which may help to exploit these biomolecules as potential novel antitumor drugs in near future.

Effect of polymer microstructure on the docetaxel release and stability of polyurethane formulation.

PurSil®AL20 (PUS), a copolymer of 4,4'-dicyclohexylmethane diisocyanate (HMDI), 1,4-butane diol (BD), poly-tetramethylene oxide (PTMO) and poly-dimethyl siloxane (PDMS) was investigated for stability as a vehicle for Docetaxel (DTX) delivery through oesophageal drug eluting stent (DES). On exposure to stability test conditions, it was found that DTX release rate declined at 4 and 40 °C. In order to divulge reasons underlying this, changes in DTX solid state as well as PUS microstructure were followed. It was found that re-crystallization of DTX in PDMS rich regions was reducing the drug release at both 4 °C and 40 °C samples. So far microstructural features have not been correlated with stability and drug release, and in this study we found that at 40 °C increase in microstructural domain sizes and the inter-domain distances (from ∼85 Å to 129 Å) were responsible for hindering the DTX release in addition to DTX re-crystallization.

Engineered atherosclerosis-specific zinc ferrite nanocomplex-based MRI contrast agents.

BACKGROUND: Cardiovascular diseases are the most prevalent cause of morbidity and mortality affecting millions of people globally. The most effective way to counter cardiovascular complications is early diagnosis and the safest non-invasive diagnostic approach is magnetic resonance imaging (MRI). In this study, superparamagnetic ferrite nanoparticles doped with zinc, exhibiting highly enhanced saturation magnetization and T2 and computed tomography (CT) contrast were synthesized. These nanoparticles have been strategically engineered using bovine lactoferrin (Lf), polyethylene glycol (PEG), and heat shock protein (Hsp)-70 antibody specifically targeting atherosclerosis with potential therapeutic value. The nanocomplexes were further validated in vitro to assess their cytotoxicity, internalization efficiency, effects on cellular proliferation and were assessed for MRI as well as X-ray CT in ex vivo Psammomys obesus rat model. RESULTS: Optimized zinc doped ferrite nanoparticles (Zn0.4Fe2.6O4) with enhanced value of maximum saturation magnetization value on 108.4 emu/g and an average diameter of 24 ± 2 nm were successfully synthesized. Successfully incorporation with bovine lactoferrin, PEG and Hsp-70 (70 kDa) antibody led to synthesis of spherical nanocomplexes (size 224.8 nm, PDI 0.398). A significantly higher enhancement in T2 (p < 0.05, 1.22-fold) and slightly higher T1 (1.09-fold) and CT (1.08-fold) contrast compared to commercial ferrite nanoparticles was observed. The nanocomplexes exhibited effective cellular internalization within 2 h in both THP-1 and Jurkat cells. MRI scans of contrast agent injected animal revealed significant arterial narrowing and a significantly higher T2 (p < 0.05, 1.71-fold) contrast in adult animals when compared to juvenile and control animals. The excised heart and aorta agar phantoms exhibited weak MRI contrast enhancement in juvenile animal but significant contrast enhancement in adult animal specifically at the aortic arch, descending thoracic aorta and iliac bifurcation region with X-ray CT scan. Histological investigation of the contrast agent injected aorta and heart confirmed site target-specific accumulation at the atherosclerotic aortic arch and descending thoracic aorta of the adult animal with severely damaged intima full of ruptured microatheromas. CONCLUSION: Overall, the study demonstrates the strategic development of nanocomplex based bimodal MRI and CT contrast agents and its validation on Psammomys obesus for atherosclerosis diagnostics.

Ultra-small algal chitosan ocular nanoparticles with iron-binding milk protein prevents the toxic effects of carbendazim pesticide.

AIM: To fabricate ultra-small algal chitosan nanoparticles (US CS NPs) for efficient delivery of bovine lactoferrin (bLf) to ocular tissues through topical administration to prevent carbendazim-induced toxicity. MATERIALS & METHODS: Rat eye model was used to evaluate the in vivo biodistribution the US CS NPs and bovine eye model was used for evaluating ex vivo biodistribution. Human lens epithelial cell line (HLEB-3) model was used to evaluate the in vitro toxicity, uptake mechanism and in vitro efficacy of the synthesized bLf-US CS NPs over carbendazim-induced ocular toxicity. RESULTS: The in vivo and ex vivo biodistribution results suggest that the ultra-small CS NPs efficiently internalize into the ocular tissues within 1 h after administering topically. Ultra-small algal nanocarriers to encapsulate bioactive antioxidant bLf protein and evaluated its potential in inhibiting carbendazim-induced human lens cell apoptosis and oxidative stress. bLf-encapsulated ultra-small algal US CS NPs prevented carbendazim-induced human lens cell apoptosis and oxidative stress. CONCLUSION: US CS NPs could be explored for their potential for delivering various ocular drugs through topical administration for other eye diseases including cataract, glaucoma and age-related macular degeneration.