Controlled Release Systems Research Articles

Development of a local controlled release system for therapeutic proteins in the treatment of skeletal muscle injuries and diseases

The present study aims to develop and characterize a controlled-release delivery system for protein therapeutics in skeletal muscle regeneration following an acute injury. The therapeutic protein, a membrane-GPI anchored protein called Cripto, was immobilized in an injectable hydrogel delivery vehicle for local administration and sustained release. The hydrogel was made of poly(ethylene glycol)-fibrinogen (PEG-Fibrinogen, PF), in the form of injectable microspheres. The PF microspheres exhibited a spherical morphology with an average diameter of approximately 100 micrometers, and the Cripto protein was uniformly entrapped within them. The release rate of Cripto from the PF microspheres was controlled by tuning the crosslinking density of the hydrogel, which was varied by changing the concentration of poly(ethylene glycol) diacrylate (PEG-DA) crosslinker. In vitro experiments confirmed a sustained-release profile of Cripto from the PF microspheres for up to 27 days. The released Cripto was biologically active and promoted the in vitro proliferation of mouse myoblasts. The therapeutic effect of PF-mediated delivery of Cripto in vivo was tested in a cardiotoxin (CTX)-induced muscle injury model in mice. The Cripto caused an increase in the in vivo expression of the myogenic markers Pax7, the differentiation makers eMHC and Desmin, higher numbers of centro-nucleated myofibers and greater areas of regenerated muscle tissue. Collectively, these results establish the PF microspheres as a potential delivery system for the localized, sustained release of therapeutic proteins toward the accelerated repair of damaged muscle tissue following acute injuries.

Green synthesized lignin nanoparticles for the sustainable delivery of pyraclostrobin to control strawberry diseases caused by Botrytis cinerea

Lignin, renowned for its renewable, biocompatible, and environmentally benign characteristics, holds immense potential as a sustainable feedstock for agrochemical formulations. In this study, raw dealkaline lignin (DAL) underwent a purification process involving two sequential solvent extractions. Subsequently, an enzyme-responsive nanodelivery system (Pyr@DAL-NPs), was fabricated through the solvent self-assembly method, with pyraclostrobin (Pyr) loaded into lignin nanoparticles. The Pyr@DAL-NPs shown an average particle size of 250.4 nm, demonstrating a remarkable loading capacity of up to 54.70 % and an encapsulation efficiency of 86.15 %. Notably, in the presence of cellulase and pectinase at a concentration of 2 mg/mL, the release of Pyr from the Pyr@DAL-NPs reached 92.66 % within 120 h. Furthermore, the photostability of Pyr@DAL-NPs was significantly improved, revealing a 2.92-fold enhancement compared to the commercially available fungicide suspension (Pyr SC). Bioassay results exhibited that the Pyr@DAL-NPs revealed superior fungicidal activity against Botrytis cinerea over Pyr SC, with an EC50 value of 0.951 mg/L. Additionally, biosafety assessments indicated that the Pyr@DAL-NPs effectively declined the acute toxicity of Pyr towards zebrafish and posed no negative effects on the healthy growth of strawberry plants. In conclusion, this study presents a viable and promising strategy for developing environmentally friendly controlled-release systems for pesticides, offering the unique properties of lignin.

Aptamer molecular gate functionalized mesoporous SiO2@MB controlled-release system for pollutant detection using Ti(Ⅲ) self-doped TiO2 NTs as active photoanode coupled with electrostatic modulation

Atrazine (ATZ) is a widely used herbicide that can cause serious harm to organisms and ecosystems. An immobilization-free photoelectrochemical (PEC) aptasensor has been herein developed for ATZ based on aptamer molecular gate functionalized mesoporous SiO2@MB controlled release system. Compared with traditional immobilization-based sensors, immobilization-free sensors (IFSs) avoid the modification of the recognition element on the electrode surface. Mesoporous SiO2 with large surface area and good biocompatibility can be used as nanocontainers to stably encapsulate the signal shuttle molecule methylene blue (MB). The bifunctional aptamer (APT) is used not only as the recognition element for ATZ but also as the signal switch to block or release MB. In the presence of ATZ, the specific recognition between ATZ and APT will cause the detachment of APT from the surface of SiO2, thus the molecular gate will open and release MB. Due to pH modulation, the positively charged MB can reach the surface of the negatively charged Ti(III) self-doped TiO2 NTs (Ti(III)–TiO2 NTs) electrode to act as an electron donor, which increases the photocurrent. The immobilization-free aptasensor has shown ultrasensitive detection of ATZ with a wide linear range from 1.0 pM to 100.0 nM and a low detection limit of 0.1 pM. In addition, the sensor has excellent selectivity, stability and anti-interference ability, and has been used in real water sample analysis successfully. This strategy has provided a new idea for the design of advanced immobilization-free PEC sensors for environmental pollutant detection.

Hydrogels in Biomedicine: Granular Controlled Release Systems Based on 2-Hydroxyethyl Methacrylate Copolymers. A Review

The article analyzes and summarizes the latest achievements in the field of polymer systems for controlled release devices based on hydrogel materials. Possible directions of drug delivery are presented, including the use of granular hydrogels, which work on the principle of drug sorption − release in the body. The research on the synthesis regularities, structure, properties, and prospects for the use of granular hydrogels based on 2-hydroxyethyl methacrylate (HEMA) and its copolymers, in particular with polyvinylpyrrolidone (PVP), as systems for the controlled release of substances, in particular, drugs, is analyzed.

Influence of Novel Microcapsulates of Bee Products on Gut Microbiota Modulation and Their Prebiotic and Pro-Adhesive Properties.

With the aim to obtain controlled-release systems and to preserve the antioxidant, immunomodulatory, and prebiotic activity of the bioactive compounds, microencapsulation of both honeydew honey and royal jelly into biopolymeric microparticles based on rye bran heteropolysaccharides (HPS) was successfully performed. Honeydew honey and royal jelly microcapsules were prepared by spray-drying method and were characterized in terms of morphology and biological properties. Due to the resistance of the obtained encapsulates to the acidic pH in the stomach and digestive enzymes, the microcapsules showed prebiotic properties positively influencing both the growth, retardation of the dying phase, and the pro-adhesive properties of probiotic bacteria, i.e., Bifidobacterium spp. and lactic acid bacteria. Moreover, as a result of fermentation of the microcapsules of bee products in the lumen of the large intestine, an increased synthesis of short-chain fatty acids, i.e., butyric acid, was found on average by 39.2% in relation to the SCFA concentrations obtained as a result of fermentation of native bee products, thus opening new perspectives for the exploitation of honeydew honey and royal jelly loaded microcapsules for nutraceutical applications.

Integrating Graphene Oxide-Hydrogels and Electrical Stimulation for Controlled Neurotrophic Factor Encapsulation: A Promising Approach for Efficient Nerve Tissue Regeneration.

Nerve growth factor (NGF) plays a crucial role in cellular growth and neurodifferentiation. To achieve significant neuronal regeneration and repair using in vitro NGF delivery, spatiotemporal control that follows the natural neuronal processes must be developed. Notably, a challenge hindering this is the uncontrolled burst release from the growth factor delivery systems. The rapid depletion of NGF reduces treatment efficacy, leading to poor cellular response. To address this, we developed a highly controllable system using graphene oxygen (GO) and GelMA hydrogels modulated by electrical stimulation. Our system showed superior control over the release kinetics, reducing the burst up 30-fold. We demonstrate that the system is also able to sequester and retain NGF up to 10-times more efficiently than GelMA hydrogels alone. Our controlled release system enabled neurodifferentiation, as revealed by gene expression and immunostaining analysis. The increased retention and reduced burst release from our system show a promising pathway for nerve tissue engineering research toward effective regeneration.

Genetic Modulators of Prion-Like Protein Propagation in Alzheimer's Disease

Alzheimer's disease (AD) is a widespread neurodegenerative disorder with complex causes. Starting with a historical overview, this paper traces the foundational discoveries by Alois Alzheimer and subsequent research into the roles of Tau, beta-amyloid (Aß), and cellular prion protein (PrPc) in AD progression. The familial aspect of AD is discussed, highlighting genetic mutations in key genes such as APP, PSEN1, PSEN2, MAPT, and APOE. Additionally, it explores epigenetic factors in familial AD and examines the prion-like behavior of Tau and Aß proteins. Potential therapeutic targets arising from these insights are considered, including PRNP manipulation and modulation of Tau and Aß behavior. A proposed stem cell therapy aims to target and counteract the pathological activities of Tau and Aß proteins through strategic genetic modifications, precise delivery to affected brain regions, and controlled release systems for sustained therapeutic effects. Ethical considerations and limitations in AD research are also addressed, emphasizing the importance of responsible research practices and equitable access to treatments. Through this exploration, the text underscores the ongoing efforts to unravel the complexities of AD and develop effective strategies for its management.

Injectable Thermosensitive Gel CH-BPNs-NBP for Effective Periodontitis Treatment through ROS-Scavenging and Jaw Vascular Unit Protection.

Periodontitis, a prevalent inflammatory condition in the oral cavity, is closely associated with oxidative stress-induced tissue damage mediated by excessive reactive oxygen species (ROS) production. The jaw vascular unit (JVU), encompassing both vascular and lymphatic vessels, plays a crucial role in maintaining tissue fluid homeostasis and contributes to the pathological process in inflammatory diseases of the jaw. This study presents a novel approach for treating periodontitis through the development of an injectable thermosensitive gel (CH-BPNs-NBP). The gel formulation incorporates black phosphorus nanosheets (BPNs), which are notable for their ROS-scavenging properties, and dl-3-n-butylphthalide (NBP), a vasodilator that promotes lymphatic vessel function within the JVU. These results demonstrate that the designed thermosensitive gel serve as a controlled release system, delivering BPNs and NBP to the site of inflammation. CH-BPNs-NBP not only protects macrophages and human lymphatic endothelial cells from ROS attack but also promotes M2 polarization and lymphatic function. In in vivo studies, this work observes a significant reduction in inflammation and tissue damage, accompanied by a notable promotion of alveolar bone regeneration. This research introduces a promising therapeutic strategy for periodontitis, leveraging the unique properties of BPNs and NBP within an injectable thermosensitive gel.

Synthesis and characterization of ion-induced sodium alginate/soy protein isolate microgels for the controlled release

In this study, sodium alginate/ soy protein isolate (SPI) microgels cross-linked by various divalent cations including Cu2+, Ba2+, Ca2+, and Zn2+ were fabricated. Cryo-scanning electron microscopy observations revealed distinctive structural variations among the microgels. In the context of gastric pH conditions, the degree of shrinkage of the microgels followed the sequence of Ca2+ > Ba2+ > Cu2+ > Zn2+. Meanwhile, under intestinal pH conditions, the degree of swelling was ranked as Zn2+ > Ca2+ > Ba2+ > Cu2+. The impact of these variations was investigated through in vitro digestion studies, revealing that all microgels successfully delayed the release of β-carotene within the stomach. Within the simulated intestinal fluid, the microgel cross-linked with Zn2+ exhibited an initial burst release, while those cross-linked with Cu2+, Ba2+, or Ca2+ displayed a sustained release pattern. This research underscores the potential of sodium alginate/SPI microgels cross-linked with different divalent cations as efficient controlled-release delivery systems.

A Color-Detectable Vitamin C Controlled-Release System Fabricated Using Electrospinning.

This study develops a vitamin C controlled-release system, trackable via color changes as a function of vitamin C release. The system is composed of coaxial microfibers prepared via coaxial electrospinning, with a core of poly(ethylene oxide) (PEO) incorporating vitamin C, and a shell composed of polycaprolactone (PCL) containing polydiacetylene (PDA) as the color-changing material. The shell thickness is controlled by adjusting the amount of PCL ejected during electrospinning, allowing regulation of the release rate of vitamin C. When vitamin C added to PEO penetrates the PCL layer, the color of PDA changes from blue to red, indicating a color change. The results of this study can be applied to devices that require immediate detection of vitamin C release levels.

Portable T-2 toxin biosensor based on target-responsive DNA hydrogel using water column height as readout

T-2 toxin, a hazardous mycotoxin often present in cereals and products based on cereals, poses a substantial risk to humans and animals due to its high toxicity. The development of uncomplicated, quick and highly sensitive methods for detecting T-2 toxin is imperative. In this work, a portable sensing system was constructed using water column height as a readout device in combination with a controlled release system, which allows for an accurate quantitative analysis of T-2 toxin without the need for expensive instrumentation or skilled technicians. Hyaluronic acid (HA) hydrogel was constructed by double cross-linked DNA/aptamer hybrids with polyethyleneimine (PEI) and embedded with platinum nanoparticles (Pt NPs). The aptamer specifically bound to T-2 toxin in its presence, resulting in the disruption of the hydrogel and subsequent release of the Pt NPs. These Pt NPs were later mixed with a solution of H2O2 in a confined reaction flask, leading to the decomposition of H2O2 into O2. A glass capillary tube containing a column of red water had been inserted into the cap of the reaction flask, and the low solubility of O2 led to an increase in pressure within the reaction unit, causing the red water column to rise. There is a good linear correlation between the height of the capillary liquid level and the T-2 toxin concentration in the range of 20 ng/mL to 6 μg/mL. The system has been successfully used to detect T-2 toxin in samples of barley tea and corn.

Advancements in microsponges for the management of vaginal and colorectal diseases: A comprehensive review

The controlled-release drug delivery systems have risen dramatically allowing various factors such as the prohibitive cost of developing new entities, the expiration of existing international patents, and the discovery of new polymeric materials suitable for prolonged drug release and improvement in therapeutic efficacy. Microsponges are the porous microspheres-based polymeric delivery system that allows controlled drug release at a specific site. Microsponges are developed for the efficient delivery of active ingredients at a low dose. They help in improving stability by modifying drug release kinetics, reducing side effects, and enhancing the retention of drug entities. Microsponge compositions are stable throughout a wide pH and temperature range, making them more compatible with numerous vehicles, and ingredients. Several studies have shown that microsponges are non-irritant, non-toxic, non-mutagenic, and non-allergic with self-sterilizing properties. They are typically used for topical administration but have lately been used for oral, vaginal, and colorectal administration as well. The current review contains basic information about microsponges, their method of preparation, and various characterization parameters. The review also discusses the application of microsponges in vaginal and colorectal diseases. The latter portion of the script includes various patents and preclinical trials.

Natural and Synthetic Polymers for Biomedical and Environmental Applications.

Natural and synthetic polymers are a versatile platform for developing biomaterials in the biomedical and environmental fields. Natural polymers are organic compounds that are found in nature. The most common natural polymers include polysaccharides, such as alginate, hyaluronic acid, and starch, proteins, e.g., collagen, silk, and fibrin, and bacterial polyesters. Natural polymers have already been applied in numerous sectors, such as carriers for drug delivery, tissue engineering, stem cell morphogenesis, wound healing, regenerative medicine, food packaging, etc. Various synthetic polymers, including poly(lactic acid), poly(acrylic acid), poly(vinyl alcohol), polyethylene glycol, etc., are biocompatible and biodegradable; therefore, they are studied and applied in controlled drug release systems, nano-carriers, tissue engineering, dispersion of bacterial biofilms, gene delivery systems, bio-ink in 3D-printing, textiles in medicine, agriculture, heavy metals removal, and food packaging. In the following review, recent advancements in polymer chemistry, which enable the imparting of specific biomedical functions of polymers, will be discussed in detail, including antiviral, anticancer, and antimicrobial activities. This work contains the authors' experimental contributions to biomedical and environmental polymer applications. This review is a vast overview of natural and synthetic polymers used in biomedical and environmental fields, polymer synthesis, and isolation methods, critically assessessing their advantages, limitations, and prospects.

Local amphotericin B therapy for Cutaneous Leishmaniasis: A systematic review.

Cutaneous leishmaniasis (CL) is characterized by potentially disfiguring skin ulcers carrying significant social stigma. To mitigate systemic drug exposure and reduce the toxicity from available treatments, studies addressing new local therapeutic strategies using available medications are coming up. This review systematically compiles preclinical and clinical data on the efficacy of amphotericin B (AmB) administered locally for cutaneous leishmaniasis. Structured searches were conducted in major databases. Clinical studies reporting cure rates and preclinical studies presenting any efficacy outcome were included. Exclusion criteria comprised nonoriginal studies, in vitro investigations, studies with fewer than 10 treated patients, and those evaluating AmB in combination with other antileishmanial drug components. A total of 21 studies were identified, encompassing 16 preclinical and five clinical studies. Preclinical assessments generally involved the topical use of commercial AmB formulations, often in conjunction with carriers or controlled release systems. However, the variation in the treatment schedules hindered direct comparisons. In clinical studies, topical AmB achieved a pooled cure rate of 45.6% [CI: 27.5-64.8%; I2 = 79.7; p = 0.002), while intralesional (IL) administration resulted in a 69.8% cure rate [CI: 52.3-82.9%; I2 = 63.9; p = 0.06). In the direct comparison available, no significant difference was noted between AmB-IL and meglumine antimoniate-IL administration (OR:1.7; CI:0.34-9.15, I2 = 79.1; p = 0.00), however a very low certainty of evidence was verified. Different AmB formulations and administration routes have been explored in preclinical and clinical studies. Developing therapeutic technologies is evident. Current findings might be interpreted as a favorable proof of concept for the local AmB administration which makes this intervention eligible to be explored in future well-designed studies towards less toxic treatments for leishmaniasis.

Multifunctional organohydrogels for on-demand controlled drug release

Controlled drug release has sparked much attention in the field of drug delivery by prolonging the duration of drug effectiveness and reducing drug toxicity. However, the development of controlled-release drug delivery systems remains a significant challenge, primarily due to inadequate drug compatibility, low biocompatibility, and low response sensitivity of the drug carriers. Herein, an innovative strategy for fabricating multifunctional organohydrogels as drug carrier systems is designed. 1-Tetradecanol/water emulsions, stabilized by cellulose acetoacetate (CAA), are initially prepared. The organohydrogels are fabricated by the ternary enzyme-mediated polymerization of the monomer- containing emulsion. The amphipathy of CAA ensures the stability of emulsions during polymerization, and CAA participates in polymerization as an initiator. Through the reversible crystallization-melting transition of 1-tetradecanol, the drug can be encapsulated in/released from the organohydrogel. Moreover, 1-tetradecanol enhances the bioavailability of water-insoluble drugs without any additional cosolvent. Therefore, biofriendly organohydrogels can serve as ideal dressings with adjustable mechanical properties, effective temperature regulation, and temperature-triggered drug release ability.

Acetazolamide encapsulation in elastin like recombinamers using a supercritical antisolvent (SAS) process for glaucoma treatment

Glaucoma, the second most common cause of blindness worldwide, requires the development of new and effective treatments. This study introduces a novel controlled-release system utilizing elastin-like recombinamers (ELR) and the Supercritical Antisolvent (SAS) technique with supercritical CO2. Acetazolamide (AZM), a class IV drug with limited solubility and permeability, is successfully encapsulated in an amphiphilic ELR at three different ELR:AZM ratios, yielding up to 62 %. Scanning electron microscopy (SEM) reveals spherical microparticles that disintegrate into monodisperse nanoparticles measuring approximately 42 nm under physiological conditions. The nanoparticles, as observed via Transmission Electron Microscopy (TEM) and Atomic Force Microscopy (AFM), do not exhibit aggregates, a fact confirmed by the zeta potential displaying a value of –33 mV over a period of 30 days. Transcorneal permeation tests demonstrate a 10 % higher permeation level compared to the control solution, which increases to 30 % after 2 h. Ocular irritation tests demonstrate no adverse effects or damage. Intraocular pressure (IOP) tests conducted on hypertensive rabbits indicate greater effectiveness for all three analyzed formulations, suggesting enhanced drug bioavailability during treatment. Consequently, the combination of recombinant biopolymers and high-pressure techniques represents a promising approach for advancing glaucoma therapy, emphasizing its potential clinical significance.

Sustained adenosine release: Revealing its impact on osteogenic signalling pathways of human mesenchymal stromal cells

Non-healing fractures, a global health concern arising from trauma, osteoporosis, and tumours, can lead to severe disabilities. Adenosine, integral to cellular energy metabolism, gains prominence in bone regeneration via adenosine A2B receptor activation. This study introduces a controlled-release system for localized adenosine delivery, fostering human mesenchymal stromal cell (hMSC) differentiation into functional bone cells. The study investigates how the ratio of lactic acid to glycolic acid in microparticles can influence adenosine release and explores the downstream effects on gene expression and metabolic profiles of osteogenic differentiation in hMSCs cultured in growth and osteoinductive media. Insights into adenosine-modulated signalling pathways during MSC differentiation, with osteogenic factors, provide a comprehensive understanding of the pathways involved. Analysing gene expression and metabolic profiles unravels adenosine's regulatory mechanisms in MSC differentiation. Sustained adenosine release from microparticles induces mineralization, synergizing with osteogenic media supplements, showcasing the potential of adenosine for treating critical bone defects and metabolic disorders. This study highlights the efficacy of a polymeric microparticle-based delivery system, offering novel strategies for bone repair. Unveiling adenosine's roles and associated signalling pathways advances our comprehension of molecular mechanisms steering bone regeneration, propelling innovative biomaterial, combined with metabolites, approaches for clinical use.

Sustained-release nitrogen fertilizer delivery systems based on carboxymethyl cellulose-grafted polyacrylamide: Swelling and release kinetics

Sustained or controlled-release delivery systems can enhance functions such as nutrient usage; minimize soil contamination, and reduce the required fertilizer dose. This paper reports the development of a carboxymethyl cellulose-g-polyacrylamide copolymer (CMC-g-PAM) as a sustained and slow-release fertilizer carrier for urea. The developed copolymer was characterized by Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Scanning Electron Microscopy (SEM), and thermo gravimetric analysis (TG). The grafting process increased the activation energy of CMC from 0.1521 to 0.5952 J/mol with a higher loading percentage of 72.140.85% using a 15% urea solution. The swelling ratio is significantly dependent on the pH. The maximum swelling ratio of 1199.58% at pH 9. However, Swelling follows a pseudo-first-order reaction with the maximum swelling ratio in a saline of 349.76%.The CMC-g-PAM copolymer loaded with urea exhibited sustained and slow release, with the maximum cumulative percentage of 69.12% at pH 9 and 38.94% in saline. Urea release from the CMC-g-PAM copolymer followed the first-order, Fickian, and biexponential biphasic release mechanisms. The release of the CMC-g-PAM copolymer loaded with urea is a complicated process governed by diffusion and a biphasic releasing profile.

Light-induced Delivery of Charged Species using Ion-selective Core-Shell Nanoparticles.

Controlled release systems have gained considerable attention owing to their potential to deliver molecules, including ions and drugs, in a customized manner. We present a light-induced ion-transfer platform consisting of a dispersion of nanoparticles (NPs, ~300 nm) with the conductive polymer poly(3-octylthiophene-2,5-diyl) (POT) in the core and a potassium (K+)-selective membrane in the shell. Owing to the photoactive nature of POT, POT NPs can be used for a dual purpose: as a host for positively charged species and as an actuator to trigger the subsequent release. POT0 and doped POT+ coexist in the core, allowing K+ encapsulation in the shell. As POT0 is photo-oxidized to POT+, K+ is released to the (aqueous) dispersion phase to preserve the neutrality of the NPs. This process is reversible and can be simultaneously assessed using the native fluorescence of POT0 and via potentiometric measurements. The NP structure and its mechanism of action were thoroughly studied with a series of control experiments and complementary techniques. Understanding the NP and its surrounding interactions will pave the way for other nanostructured systems, facilitating sophisticated applications. The delivery of ionic drugs and interference/pollutant catching for advanced sensing/restoration will be considered in future research.

Isoliquiritigenin-infused electrospun nanofiber inhibits breast cancer proliferation and invasion through downregulation of PI3K/Akt/mTOR and MMP2/9 pathways

Mastectomy followed by chemotherapy and radiotherapy is the current treatment regimen for breast cancer (BC). However, these approaches have systemic side effects and often lead to ineffective treatment outcomes. Therefore, a novel controlled release system is required to prevent tumor recurrence. Isoliquiritigenin (ISL), a dietary flavonoid, was shown to be effective against BC in our laboratory. However, its hydrophobicity and low bioavailability limit its anti-BC efficacy. Thus, the present study was designed to fabricate electrospun ISL-nanofiber scaffolds (ISL-NF) that can inhibit breast cancer growth and migratory capacity of BC cells in vitro and in vivo. The ISL-NF was prepared using an advanced coaxial electrospinning-cryocutting method, and characterized. Various assays, including cell viability, transwell, would healing, western blotting, and staining, were employed to explore the underlying mechanisms. In vitro studies showed that ISL-NF treatment for 24 h significantly inhibited cell proliferation, and migration and improved apoptosis in various BC cell lines. ISL-NF also decreased the protein expression of PI3K, Akt, mTOR, matrix metalloproteinase (MMP)-2, and MMP-9, thereby inhibiting BC cell invasion. Consistent with the in vitro study, ISL-NF treatment decreased tumor size and weight and inhibited tumor migration in 4T1 tumor-bearing mice. Based on the study, we suggest that ISL-NF may be useful in the treatment of BC.