Nano-sized Liposomes Research Articles

Exploring the influence of microstructure and phospholipid type of liposomes on their interaction with lung

Liposome is a promising carrier for pulmonary drug delivery and the nano-sized liposomes have been widely investigated in the treatment of lung diseases. However, there still lack the knowledge of micron-sized liposomes for lung delivery, which have more advantages in terms of drug loading and sustained drug release capacity. The micron-sized liposomes can be classified into multilamellar liposome (MLL) and multivesicular liposome (MVL) according to their microstructure, thus, this study focused on exploring how the micron-sized liposomes with different microstructure and phospholipid composition influence their interaction with the lung. The MLL and MVL were prepared from different types of phospholipids (including soya phosphatidylcholine (SPC), egg yolk phosphatidylcholine (EPC), and dipalmitoyl phosphatidylcholine (DPPC)) with geometric diameter around 5 μm, and their in vitro pulmonary cell uptake, in vivo lung retention and organ distribution were investigated. The results showed that the microstructure of liposomes didn’t affect pulmonary cellular uptake, in vivo lung retention and organ distribution. MLL and MVL prepared with the same phospholipid had similar cellular uptake in both NR8383 cells and A549 cells, and both of them possessed prolonged lung retention and limited distribution in other organs during 72 h. Notably, the phospholipid type presented remarkable influence on liposomes’ interaction with the lung. SPC-based liposomes exhibited higher cellular uptake than the DPPC-based ones in both NR8383 cells and A549 cells, also possessed a better lung retention behavior. In conclusion, this study might provide theoretical knowledge for designing micron-sized liposomes intended for lung delivery.

Synthesis of α,ω-bis-Mercaptoacyl Poly(alkyl oxide)s and Development of Thioether Cross-Linked Liposome Scaffolds for Sustained Release of Drugs.

With the aim to develop novel scaffolds for the sustained release of drugs, we initially developed an easy approach for the synthesis of α,ω-homobifunctional mercaptoacyl poly(alkyl oxide)s. This was based on the esterification of the terminal hydroxyl groups of poly(alkyl oxide)s with suitably S-4-methoxytrityl (Mmt)-protected mercapto acids, followed by the removal of the acid labile S-Mmt group. This method allowed for the efficient synthesis of the title compounds in high yield and purity, which were further used in the development of a thioether cross-linked liposome scaffold, by thia-Michael reaction of the terminal thiol groups with pre-formed nano-sized liposomes bearing maleimide groups on their surface. The reaction process was followed by 1H-NMR, using a Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion NMR experiment (1H-NMR CPMG), which allowed for real-time monitoring and optimization of the reaction process. The thioether cross-linked liposomal scaffold that was synthesized was proven to preserve the nano-sized characteristics of the initial liposomes and allowed for the sustained release of calcein (which was used as a hydrophilic dye and a hydrophilic drug model), providing evidence for the efficient synthesis of a novel drug release scaffold consisting of nanoliposome building blocks.

Determination of Protein Amount in Nanosized Synthetic Liposomes by Surface Effect Raman Spectroscopy (SERS)

Accurate characterization of synthetic liposomes is essential since they give information about the vesicu-lar structures in bodily fluids such as extracellular vesicles. The characterization tasks are generally the determination of the sizes of the liposomes and the profiling of the liposomes' content. Optical tweezers and Surface Enhanced Raman Spectroscopy (SERS) were used to profile the nanosized liposomes. The size distribution of the trapped liposomes (140 nm on average) was found by using Einstein's Brownian motion equation, consistent with the size distribution obtained from dynamic light scattering measure-ments. Besides, Gramicidin-encapsulated liposomes were measured using SERS, and statistically signifi-cant differentiation was found in Raman intensities between liposome populations with altering concentra-tions of proteins. This study uniquely measured size distributions of nano-sized liposomes with conven-tional optical tweezers (without plasmonics) and determined the chemical differences between empty and protein encapsulated liposomes with high accuracy using Raman spectroscopy

Expansion supercritical fluid into an aqueous solution (ESSAS), a new technique for creating nano-size cyanocobalamin-loaded liposomes, and optimization of involved parameters

Expansion supercritical fluid into an aqueous solution (ESSAS), a new technique for creating nano-size cyanocobalamin-loaded liposomes, and optimization of involved parameters

68 Ga-labeled, imatinib encapsulated, theranostic liposomes: Formulation, characterization, and in vitro evaluation of anticancer activity.

Cancer is still a global health problem. Among cancer types, breast cancer is the most frequently diagnosed one, and it causes a high mortality rate if not diagnosed in the early stages. In our study, imatinib encapsulated, nanosized, neutral/cationic liposome formulations were prepared as theranostic agents for breast cancer. After the characterization studies in which all liposomes exhibited proper profile owing to their particle size between 133 and 250 nm, polydispersity index valueslower than 0.4, neutral and cationic zeta potential values, and high drug encapsulation efficiency, controlled drug release behaviors with zero-order kinetic were obtained. The higher than 90% radiolabeling efficiency values were obtained thanks to the determination of optimum radiolabeling condition (80°C temperature, 5 mCi radioactivity, and 10 min incubation period). According to the resazurin assay evaluating the cytotoxic profile of liposomes on MCF7 cells, neutral empty liposome was found as biocompatible, while both cationic liposomes (empty and drug-loaded ones) exhibited high nonspecific cytotoxicity at even low drug concentration due to the existence of stearyl amine in the formulations. However, dose-dependent cytotoxic effect and the highest cellular binding capacity were obtained by imatinib loaded neutral liposomes. In conclusion, 68 Ga-radiolabeled, imatinib-loaded, neutral, nanosized liposome formulation is the most promising one as a theranostic agent among all formulations.

Preparation of Uniform Nano Liposomes Using Focused Ultrasonic Technology.

Liposomes are microspheres produced by placing phospholipids in aqueous solutions. Liposomes have the advantage of being able to encapsulate both hydrophilic and hydrophobic functional substances and are thus important mediators used in cosmetics and pharmaceuticals. It is important for liposomes to have small sizes, uniform particle size distribution, and long-term stability. Previously, liposomes have been prepared using a homo mixer, microfluidizer, and horn and bath types of sonicators. However, it is difficult to produce liposomes with small sizes and uniform particle size distribution using these methods. Therefore, we have developed a focused ultrasound method to produce nano-sized liposomes with better size control. In this study, the liposome solutions were prepared using the focused ultrasound method and conventional methods. The liposome solutions were characterized for their size distribution, stability, and morphology. Results showed that the liposome solution prepared using focused ultrasonic equipment had a uniform particle size distribution with an average size of 113.6 nm and a polydispersity index value of 0.124. Furthermore, the solution showed good stability in dynamic light scattering measurements for 4 d and Turbiscan measurements for 1 week.

M2 Macrophage Hybrid Membrane-Camouflaged Targeted Biomimetic Nanosomes to Reprogram Inflammatory Microenvironment for Enhanced Enzyme-Thermo-Immunotherapy.

Excessive inflammatory reactions caused by uric acid deposition are the key factor leading to gout. However, clinical medications cannot simultaneously remove uric acid and eliminate inflammation. An M2 macrophage-erythrocyte hybrid membrane-camouflaged biomimetic nanosized liposome (USM[H]L) is engineered to deliver targeted self-cascading bienzymes and immunomodulators to reprogram the inflammatory microenvironment in gouty rats. The cell-membrane-coating endow nanosomes with good immune escape and lysosomal escape to achieve long circulation time and intracellular retention times. After being uptaken by inflammatory cells, synergistic enzyme-thermo-immunotherapies are achieved: uricase and nanozyme degraded uric acid and hydrogen peroxide, respectively; bienzymes improved the catalytic abilities of each other; nanozyme produced photothermal effects; and methotrexate has immunomodulatory and anti-inflammatory effects. The uric acid levels markedly decrease, and ankle swelling and claw curling are effectively alleviated. The levels of inflammatory cytokines and ROS decrease, while the anti-inflammatory cytokine levels increase. Proinflammatory M1 macrophages are reprogrammed to the anti-inflammatory M2 phenotype. Notably, the IgG and IgM levels in USM[H]L-treated rats decrease substantially, while uricase-treated rats show high immunogenicity. Proteomic analysis show that there are 898 downregulated and 725 upregulated differentially expressed proteins in USM[H]L-treated rats. The protein-protein interaction network indicates that the signaling pathways include the spliceosome, ribosome, purine metabolism, etc.

Evaluation of efficacy of tumor-specific nanoliposomal radiosensitizer in radiotherapy

Radiotherapy is one of the cancer treatment options in which ionizing radiation is used. While the ionizing radiation used here damages the tumor cells, it also damages the surrounding healthy cells and tissues. One of the approaches used to reduce the undesirable side effects and increase the therapeutic efficacy of radiotherapy is the use of radiosensitizers. Up to now, different radiosensitizers have been investigated for this purpose and recently, nanotechnology-based radiosensitizer research has attracted much attention. In this study, the therapeutic efficacy of tumor-targeted, nano-sized liposomal radiosensitizers containing quantum dots-photosensitizer conjugate was evaluated in vivo. It was aimed to destroy more tumor cells by using same the radiation doses routinely used in cancer treatment in radiation oncology clinics or to destroy about the same amount of tumor cells by using lower radiation doses than the routinely used clinical dose. For this purpose, nano-sized, PEG-coated, and folic acid-modified tumor-specific liposomes loaded with quantum dots (QD = CdSe/ZnS) - photosensitizer (Ce6 = Clorine-e6) conjugate was prepared and their therapeutic efficacy was evaluated in 4T1 murine breast cancer cell tumor-bearing mice. Following the single dose of 10Gy 6 MV X-ray irradiation after administration of liposomal radiosensitizer, changes in animals' weights and tumor volumes were measured and animal survival was monitored to evaluate the efficacy of the treatment. Histopathological studies were also performed to examine possible damage to the tumor and normal tissues. In control groups, only X-ray irradiation was applied or no treatment was applied to mice. The results showed that the liposomal radiosensitizer plus radiotherapy killed more cancer cells than radiotherapy alone. While tumor volumes increased in the control group, approximately a 39% reduction in tumor volumes was observed in mice treated with x-ray irradiation following administration of the liposomal radiosensitizer. The obtained results showed that nano-sized liposomal radiosensitizers could be a promising radiosensitizer to achieve better treatment efficacy in radiotherapy by applying fewer radiation doses compared to clinically used radiation doses.

Development of galangin-loaded nano-sized polyelectrolyte liposome: optimization and characterization

Development of galangin-loaded nano-sized polyelectrolyte liposome: optimization and characterization

Abstract 831: Selective targeting of pediatric rhabdoid tumor using G401-CLENs: Cytotoxicity studies

Abstract Background: Rhabdoid tumor is an aggressive soft tissue sarcoma that arises in the kidney, liver, peripheral nerves, and miscellaneous soft parts throughout the body. Rhabdoid tumors usually occur in infancy or during early childhood and the most common sites for metastasis are lungs, abdominal lymph nodes, liver, and brain. Current treatment options include surgery, radiation therapy, high-dose chemotherapy, and stem cell transplant but the overall survival rate is approximately 23%. Nano-sized liposomes are drug-delivery vehicles used to enhance the delivery of drugs to target cells. The composition, size, and overall charge of nanoliposomes determine targeting and influence the degree of drug uptake. The objective of this study was to develop G401-CLENs prepared from lipid extracts derived from rhabdoid tumor cells and conventional lipid components, and to evaluate the optimized drug loaded preparations of rhabdoid-CLENs in vitro. Methods: Rhabdoid tumor cell line, G401, was the chosen model for this study. G401 cells were cultured and expanded in vitro for cellular extraction purposes. The G401 cell membrane lipid extract (LE) was included in the preparation of G401-CLENs. Additional conventional lipid ingredients include DOPC, DOTAP, and DPPE-PEG5000. DPPE-rhodamine lipids were employed for fluorescence studies only. Chemo agents' doxorubicin and vinblastine were used in cell viability and cytotoxicity assays to evaluate formulation efficacy. Phase I of evaluation included physiochemical characterization and cellular uptake studies of preparations of CLENs, including 0 to 20 mol% of G401 LE content. Phase II of evaluation included the use of Hoechst33342/propidium iodide cell viability and cytotoxicity assays with varied amounts of doxorubicin and vinblastine loaded CLENs added to the G401 cells. The control cell line used in the study represents another solid tumor, SK-N-DZ (neuroblastoma). Results: The optimal composition for G401-CLENs was DOPC/DPPE-PEG5000/LE (93/2/5). The average size and zeta potential was 198 ± 16 nm (n=5) and -12 ± 1.2 mV (n=5), respectively. Cell selectivity studies revealed an increase in the uptake of the optimized G401-CLENs by G401 cells when compared to SK-N-DZ control cells. In general, the inclusion of DOTAP increased overall cellular uptake, but diminished selective uptake of G401-CLENs. Results support a significant cellular uptake of preparations consisting of 5 mol% LE compared to 0, 10, and 20 mol% G401 LE content. Doxorubicin loaded in G401-CLENs decreased rhabdoid tumor cell viability in a dose-dependent manner compared to doxorubicin loaded in conventional liposome preparations. Additional cytotoxicity studies are currently underway. Conclusion: To this date, G401-CLENs represents a promising selective drug-targeting vehicle for the treatment of rhabdoid tumors. Citation Format: Varun Kudchadker, Dongxiao Chen, Yeji Seo, Robert Campbell. Selective targeting of pediatric rhabdoid tumor using G401-CLENs: Cytotoxicity studies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 831.

Precise control of liposome size using characteristic time depends on solvent type and membrane properties

Controlling the sizes of liposomes is critical in drug delivery systems because it directly influences their cellular uptake, transportation, and accumulation behavior. Although hydrodynamic focusing has frequently been employed when synthesizing nano-sized liposomes, little is known regarding how flow characteristics determine liposome formation. Here, various sizes of homogeneous liposomes (50–400 nm) were prepared according to flow rate ratios in two solvents, ethanol, and isopropyl alcohol (IPA). Relatively small liposomes formed in ethanol due to its low viscosity and high diffusivity, whereas larger, more poly-dispersed liposomes formed when using IPA as a solvent. This difference was investigated via numerical simulations using the characteristic time factor to predict the liposome size; this approach was also used to examine the flow characteristics inside the microfluidic channel. In case of the liposomes, the membrane rigidity also has a critical role in determining their size. The increased viscosity and packing density of the membrane by addition of cholesterol confirmed by fluorescence anisotropy and polarity lead to increase in liposome size (40–530 nm). However, the interposition of short-chain lipids de-aligned the bilayer membrane, leading to its degradation; this decreased the liposome size. Adding short-chain lipids linearly decreased the liposome size (130–230 nm), but at a shallower gradient than that of cholesterol. This analytical study expands the understanding of microfluidic environment in the liposome synthesis by offering design parameters and their relation to the size of liposomes.

Nucleotides Entrapped in Liposome Nanovesicles as Tools for Therapeutic and Diagnostic Use in Biomedical Applications.

The use of nucleotides for biomedical applications is an old desire in the scientific community. As we will present here, there are references published over the past 40 years with this intended use. The main problem is that, as unstable molecules, nucleotides require some additional protection to extend their shelf life in the biological environment. Among the different nucleotide carriers, the nano-sized liposomes proved to be an effective strategic tool to overcome all these drawbacks related to the nucleotide high instability. Moreover, due to their low immunogenicity and easy preparation, the liposomes were selected as the main strategy for delivery of the mRNA developed for COVID-19 immunization. For sure this is the most important and relevant example of nucleotide application for human biomedical conditions. In addition, the use of mRNA vaccines for COVID-19 has increased interest in the application of this type of technology to other health conditions. For this review article, we will present some of these examples, especially focused on the use of liposomes to protect and deliver nucleotides for cancer therapy, immunostimulatory activities, enzymatic diagnostic applications, some examples for veterinarian use, and the treatment of neglected tropical disease.

A delivery system into the dermis layer of skin using small unilamellar liposomes and targeted collagen regeneration stimulated by electric field

Collagen is one of the prominent scaffolding components of the extracellular matrix that can turn into collagen fibers through self-assembly mechanisms. Understanding that various factors can influence collagen assembly, we studied the effect of electric field exposure and visualized collagen fibrils' fibrillogenesis process under the confocal microscope. Collagen monomers started to aggregate within 10 seconds after applying an AC of 10 Hz-1V, and collagen fibrils with a 28- to 70-nm diameter appeared within 9 minutes. Further, we designed an experiment to form collagen fibers at a targeted region by encapsulating collagen molecules in nano-sized liposomes (Lip-Col) and have confirmed that the fibrils were successfully formed. We infer that the electric field could trigger the electrical interaction between hydronium ion and the hydroxyl group of hydroxyproline in the collagen peptide chains, resulting in the change of the surface charge of the collagen and pH solution. In vitro, Lip-Cols can interact with the cellular membrane of human dermal fibroblast and form collagen networks in extracellular matrix space. Our findings can potentially be utilized as a drug delivery system for anti-aging or wound healing agents.

Reactive Oxygen Species-Enhanced Low-Dose Chemo-Photodynamic Therapy for Gastric Cancer

A photosensitizer can react with oxygen or a substrate based on energy or electron transfer processes in order to generate free radicals, singlet oxygen (SOG), or other reactive oxygen species (ROS). SOG can damage cellular components and eventually cause cell death. However, SOG generation requires sufficient oxygen. Therefore, the effect of photodynamic therapy (PDT) via SOG may be decreased in oxygen-deficient tumor tissues, including gastric cancer. Here, we synthesized a nanosized cationic liposome integrated with a photosensitizer and low-dose chemotherapeutic drug related to ROS to improve the ROS-mediated PDT effect and lower the risk of drug resistance and side effects induced by chemotherapy. The cationic liposome was used as a drug carrier to increase the efficiency of drug delivery, prolong drug circulation time, and achieve the simultaneous delivery of the photosensitizer and chemotherapeutic drug. The low-dose chemotherapeutic agent related to ROS was adopted for increasing the permeability of the mitochondrial membrane and inducing the entry of high levels of the photosensitizer into the cell so as to generate toxic ROS. Generally, the ROS-mediated enhanced effect could quickly induce long-lasting apoptosis. This integrated nanosized cationic liposome system is a potential agent for gastric cancer therapy and is based on enhanced ROS-mediated synergistic therapy.

A comparative study on micelles, liposomes and solid lipid nanoparticles for paclitaxel delivery

The purpose of this work was to compare the in vitro and in vivo characteristics of LDV-targeted lipid-based micelles, liposomes and solid lipid nanoparticles (SLN) to provide further insights into their therapeutic potential for clinical development. Micelles, liposomes and SLN were prepared using LDV peptide amphiphiles and palmitic acid-derived lipids using solvent evaporation, thin-film hydration and microfluidic mixing respectively. Nanocarriers were characterized for their physicochemical properties, paclitaxel loading efficiency, in vitro release behavior, stability in biological media as well as in vivo antitumor efficacy in melanoma xenograft model. TEM and DLS results confirmed the presence of paclitaxel-loaded nanosized micelles (6 to 12 nm), liposomes (123.31 ± 5.87 nm) and SLN (80.53 ± 5.37 nm). SLN demonstrated the slowest paclitaxel release rate and the highest stability in biological media compared to micelles and liposomes. Paclitaxel-loaded SLN demonstrated a statistically significant delay in tumor growth compared to mice treated with paclitaxel-loaded liposomes and paclitaxel-loaded micelles (p < 0.05). The results obtained in this study indicate the potential of SLN as drug delivery vehicles for anticancer therapy.

Recent Approaches and Success of Liposome-Based Nano Drug Carriers for the Treatment of Brain Tumor.

Brain tumors are nothing but a collection of neoplasms that originate either from areas within the brain or from systemic metastasized tumors of other organs spread to the brain. It is a leading cause of death worldwide. The presence of the blood-brain barrier (BBB), blood-brain tumor barrier (BBTB), and some other factors may limit the entry of many potential therapeutics into the brain tissues in the tumor area at the therapeutic concentration required for satisfying effectiveness. Liposomes play an active role in delivering many drugs through BBB into the tumor due to their nanosize and physiological compatibility. The surface of the liposomes can be modified with various ligands that are very specific to the numerous receptors overexpressed onto the BBB as well as onto the diseased tumor surface site (i.e., BBTB) to deliver selective drugs into the tumor site. Further, this colloidal carrier can encapsulate both lipophilic and hydrophilic drugs due to its unique structure. Moreover, the enhanced permeability and retention (EPR) effect can be an added advantage for nanosize liposomes to concentrate into the tumor microenvironment through relatively leaky vasculature of solid tumor in the brain where no penetration restriction applies compared to normal BBB. Here in this review, we have tried to compile the recent advancement along with the associated challenges of liposomes containing different anti-cancer chemotherapeutics across the BBB/BBTB for the treatment of gliomas that will be very helpful for the readers for better understanding of different trends of brain tumor targeted liposomes-based drug delivery and for pursuing fruitful research on the similar research domain.

Aptamer-conjugated nano-liposome for immunogenic chemotherapy with reversal of immunosuppression.

Cancer cells have various immune evasion mechanisms that resist the immune cells by reprogramming the tumor microenvironment (TME), such as programmed death-ligand 1 (PD-L1) and indoleamine 2,3-dioxygenase-1 (IDO1) overexpression. One of the approaches to restore antitumor immune response by T-cells is through induction of immunogenic cell death (ICD). Thus, drug carrier containing IDO1 siRNA and ICD inducer would be effective anticancer regimen to modulate the immunosuppressive TME by reversing the IDO1-mediated immunosuppression in a synergistic combination with ICD induction. However, numerous nanocarrier platforms for co-delivery of multiple drugs mostly depend on the enhanced permeation and retention (EPR), which is insufficient to achieve selectivity in tumor sites harboring various types of cells. We designed a targeted drug delivery system using nano-sized liposomes functionalized with anti-CD44 and anti-PD-L1 DNA aptamers, which target breast cancer cells and inhibit PD-1/PD-L1 interaction between cancer cells and T-cells. To reverse immunosuppressive TME and reactivate immune response, cancer-targeting nano-liposomes were prepared to contain immunogenic cell death inducer (Doxorubicin, DOX) and IDO1 siRNA, namely Aptm[DOX/IDO1]. The Aptm[DOX/IDO1] specifically delivered the loaded DOX and IDO1 siRNA into target breast cancer cells through aptamer-mediated endocytosis. Cancer-targeted DOX/IDO1 siRNA delivery enhanced ICD and suppressed IDO1 expression with significantly high toxicity in cancer cells. We demonstrated that Aptm[DOX/IDO1] could achieve synergistic antitumor effects by facilitating ICD response and simultaneous reversal of the immunosuppressive TME with IDO1 knockdown in the subcutaneous breast cancer model mice, thus reducing tumor size. These antitumor effects were exerted with intratumoral infiltration of CD8+ cytotoxic T lymphocyte as well as attenuation of regulatory T-cell recruitment in the tumor sites. We further proved that our Aptm[DOX/IDO1] strategy significantly reduced tumor metastasis in tumor-xenograft mice through a synergistic combination of cancer cell-targeted ICD induction and reversal of the IDO1-mediated immunosuppressive TME. Our nanocarrier platform based on cationic liposomes containing DOX and IDO1 siRNA, which are conjugated with two DNA aptamers targeting the cancer cell surface, accomplished synergistic chemoimmunotherapy through tumor-specific immune modulation into immune-favorable TME in vivo.

Composite Norland Optical Adhesive (NOA)/silicon flow focusing devices for colloidal particle manipulation and synthesis

Microfluidic flow focusing devices are widely used to generate steep chemical concentration gradients at the interface between miscible or partially miscible streams. In this study, first we present an optimised protocol for the manufacturing of composite flow focusing devices, consisting of a micropatterned layer of Norland Optical Adhesive (NOA) glue bound to flat or microgrooved silicon substrates. Then, three different applications of these devices are demonstrated, namely (i) particle spreading and focusing in continuous flows past flat substrates, (ii) particle accumulation within the dead-end cavities of microgrooved substrates and (iii) synthesis of nano-sized liposomes. Colloidal particle spreading, focusing and accumulation were achieved through diffusiophoresis transport induced by salt concentration gradients at the interface between electrolyte streams. Epi-fluorescence microscopy was adopted to characterise the spatio-temporal distribution of silica and polystyrene nanoparticles in the devices with flat or microgrooved surfaces. The effects of particle zeta potential and groove thickness on particle dynamics were investigated. 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine (DOPC) liposomes were generated by hydrodynamic focusing and characterised via dynamic light scattering. Liposome populations with controlled narrow size distributions could be achieved by adjusting the flow rate conditions in the devices. This work demonstrates that composite NOA/silicon flow junction devices offer a competitive alternative to conventional PDMS chips and can support a wide range of microfluidic applications, including nanoparticle synthesis, characterisation and filtration, drug encapsulation and biochemical analysis.

Quality-by-design-based engineered liposomal nanomedicines to treat cancer: an in-depth analysis.

Engineered nano-sized liposomes have attained the highest success rate in commercialization among the reported nanomedicines. However, developing industrially acceptable nanoliposomes is still challenging because the process, formulation factors and even their properties may critically influence the desired attributes of the final nanoliposomal product. Implementation of quality-by-design (QbD) in nanoliposomal fabrication has led to revolutionary advancement int better analysis of the interacting factors (drug and lipid ratio, hydration, sonication, etc), which, in turn, leads to better product performance with predefined attributes (entrapment efficiency percentage, drug release time and pattern, vesicles size, polydispersity index, surface chargeand surface morphology). This review provides a summary of decade of researchand an in-depth analysis of QbD-based nanoliposomes developed to address different cancers. The review aims to provide complete details of QbD-inspired nanoliposomal development from process to application.

Abstract 306: Selective targeting of pediatric acute lymphoblastic leukemia using CCRF-CEM CLENs: Development and characterization

Abstract Background: Acute lymphoblastic leukemia (ALL) is a hematological malignancy commonly observed in children under 14 years old. A defect in the hematopoietic pathway results in the uncontrolled proliferation of B-cell and T-cell lymphocyte precursors. Treatments for ALL are patient-specific and are determined based on prognostic factors such as age, minimal residue disease, and genetic alterations. Nano-sized liposomes are drug-delivery vehicles used to enhance the delivery of drugs to target cells. The composition, size, and overall charge of nanoliposomes are determinants of targeting and influence the degree of drug uptake selectivity. The objective of this study was to design, optimize, and characterize novel cell membrane lipid-extracted nanoliposomes (CLENs) for pediatric ALL and to evaluate the degree of selectivity in comparison to relevant controls. Methods: The ALL-cell line, CCRF-CEM, was the chosen model for ALL disease and drug selectivity studies. CCRF-CEM cells were cultured and expanded in vitro for cellular extraction purposes. The CCRF-CEM cell lipid extract (LE) was included to prepare CCRF-CEM CLENs. Phase I of evaluation included physiochemical characterization and cellular uptake studies of preparations of CLENs, including 0 to 40 mol% of CCRF-CEM LE content. Phase II of evaluation fixed the optimal LE content determined from the Phase I studies and varied the cholesterol (Chol) content (0 to 40 mol%). The control cell lines used in the study represent other hematological diseases, RPMI 8226 (multiple myeloma), K-562-GFP (chronic myeloid leukemia), and U937 (lymphoma). Conventional lipid ingredients used in preparing CCRF-CEM-CLENs include DOPC, Chol, and DPPE-rhodamine for fluorescence studies using a fluorescence microplate reader. Results: The optimal composition (and ratio of components) for CCRF-CEM CLENs was DOPC/Chol/LE (85/10/5). Results support a significant uptake of preparation types consisting of 0% LE compared to 5% LE content (p=0.01). A similar effect was observed between 0% and 10% Chol content (p&amp;lt;0.001). The steady inclusion of Chol (2, 5, 10%) increased the cellular uptake of CCRF-CEM CLENs by CCRF-CEM cells, above which the cellular uptake decreased. The average mean diameter for the optimized preparation type was 182 ± 44 nm (n=5). The average zeta potential was -9 ± 6 mV (n=5). Cell selectivity studies revealed an increase in the uptake of the optimized CCRF-CEM CLENs by CCRF-CEM cells when compared to U937 (p=0.002) and RPMI 8226 (p=0.002) control cells. No statistical significance was observed in uptake of the same preparations by CCRF-CEM, when compared to K-562-GFP cells (p=0.09). Conclusion: ALL-CLENs has demonstrated promising preliminary results. Future studies will investigate mechanisms underlying formulation and cellular properties. Citation Format: Andre Yonan, Christopher Jacques, Thanaphorn Feinberg, Tafaswa Fletcher, Robert Campbell. Selective targeting of pediatric acute lymphoblastic leukemia using CCRF-CEM CLENs: Development and characterization [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 306.