Potential Of Nanostructured Lipid Carriers Research Articles

Copaifera langsdorffii Oleoresin-Loaded Nanostructured Lipid Carrier Emulgel Improves Cutaneous Healing by Anti-Inflammatory and Re-Epithelialization Mechanisms.

The skin is essential to the integrity of the organism. The disruption of this organ promotes a wound, and the organism starts the healing to reconstruct the skin. Copaifera langsdorffii is a tree used in folk medicine to treat skin affections, with antioxidant and anti-inflammatory properties. In our study, the oleoresin of the plant was associated with nanostructured lipid carriers, aiming to evaluate the healing potential of this formulation and compare the treatment with reference drugs used in wound healing. Male Wistar rats were used to perform the excision wound model, with the macroscopic analysis of wound retraction. Skin samples were used in histological, immunohistochemical, and biochemical analyses. The results showed the wound retraction in the oleoresin-treated group, mediated by α-smooth muscle actin (α-SMA). Biochemical assays revealed the anti-inflammatory mechanism of the oleoresin-treated group, increasing interleukin-10 (IL-10) concentration and decreasing pro-inflammatory cytokines. Histopathological and immunohistochemical results showed the improvement of re-epithelialization and tissue remodeling in the Copaifera langsdorffii group, with an increase in laminin-γ2, a decrease in desmoglein-3 and an increase in collagen remodeling. These findings indicate the wound healing potential of nanostructured lipid carriers associated with Copaifera langsdorffii oleoresin in skin wounds, which can be helpful as a future alternative treatment for skin wounds.

Potential of nanostructured lipid carriers in oral delivery of the poorly soluble drugs

Potential of nanostructured lipid carriers in oral delivery of the poorly soluble drugs

In vitro and in vivo characterization of Entacapone-loaded nanostructured lipid carriers developed by quality-by-design approach

Entacapone, a reversible catechol-o-methyl transferase inhibitor, is used to enhance the action of dopamine agonists by reducing their metabolism and the ‘Wearing-off’ effects associated with long-term use in the treatment of Parkinson's disease. It is used as an adjunct to levodopa/Carbidopa therapy. Due to limited dissolution and first-pass clearance, it suffers low and variable bioavailability issues. To overcome this problem, the present study aims to explore the potential of nanostructured lipid carriers (NLCs) for the delivery of Entacapone. The Quality by Design (QbD) approach was used for the systematic development of NLCs. The 23 full factorial designs were investigated using Design-Expert®11 software. The three independent variables namely content of total lipid (X1), surfactant (X2), and sonication time (X3) were optimized against two responses namely particle size and entrapment efficiency. The optimized NLCs were characterized for their size, surface morphology, entrapment efficiency, drug release, thermal and crystallographic studies. In-vivo pharmacokinetic studies in Entacapone-loaded NLCs showed an increase in t 1/2, AUC0–∞, MRT compared to free drug. The reduction in elimination (Kel) depicts the prolonged action of Entacapone by loading in NLCs. The results displayed Entacapone-loaded NLCs have promising potential for oral delivery and enhanced therapeutic effect which otherwise was a major issue.

Fabrication and characterization of nanostructured lipid carrier system for effective delivery of poorly water-soluble drug quetiapine fumarate

The aim of present study is to investigate the potential of nanostructured lipid carriers (NLCs) in improving the oral bioavailability of quetiapine fumarate, a second-generation antipsychotic drug. Quetiapine Fumarate (QF) loaded NLC were prepared by hot homogenization followed by an ultrasonication method. Response surface methodology - central composite design (CCD) was used to systemically examine the influence of concentration of capmul MCM EP, concentration of poloxamer 188 and concentration of egg lecithin on particle size (PS) and % entrapment efficiency (% EE) and to optimize the NLC formulation. The CCD consists of three factored design with five levels, plus and minus alpha (axial points), plus and minus 1 (factorial points) and the centre point. A mathematical relationship between variables was created by using Design Expert software Version 12. The statistical evaluations revealed that three independent variables were the important factors that affected the PS and % EE of QF loaded NLC. The best fitted mathematical model was linear and quadratic for PS and % EE respectively. The optimized formulations found with 218.1±0.14nm of PS and 93±0.16% of % EE. Results illustrated the superiority of developed QF loaded NLC formulation as a stable drug delivery system, providing better bioavailability with the possibility of better treatment for psychological disorders.

Multi-organ targeting of HIV-1 viral reservoirs with etravirine loaded nanostructured lipid carrier: An in-vivo proof of concept

The inadequate bioavailability and toxicity potential of antiretroviral therapy limit their effectiveness in the complete eradication of HIV from viral reservoirs. The penetration of these drugs into the brain is challenging because of the unfavorable physicochemical properties required to cross the membranes, limiting the transport of the drugs. Thus, in the current study, the authors report a nanocarrier-based drug delivery of a highly hydrophobic drug to overcome the existing limitations of the conventional therapies. An explicitly simple approach was used to overcome the limitations of existing anti-HIV therapies. The monophasic hot homogenized solution of lipid, drug, and solubilizer was diluted with the predetermined hot surfactant solution followed by the ultrasonication to generate the polydisperse nanoparticles with the size range of 50-1000 nm. The anti-HIV1 potential of nanostructured lipid carriers of Etravirine on HIV-infected cell lines showed efficacy with an appreciable increase in the therapeutic index as compared with the plain drug. Further, the results obtained from confocal microscopy along with flow cytometry exhibited efficient uptake of the nanocarrier loaded with coumarin-6 in cells. The pharmacokinetics of Etravirine nanostructured carriers was significantly better in all aspects compared to the plain drug solution, which could be attributed to molecular dispersion in the lipid matrix of the nanocarrier. A significant enhancement of Etravirine concentration of several-fold was also observed in the liver, ovary, lymph node, and brain, respectively, as compared to plain drug solution when assessed by biodistribution studies in rats. In conclusion, ETR-NLC systems could serve as a promising approach for simultaneous multi-site targeting and could provide therapeutic benefits for the efficient eradication of HIV/AIDS infections.

Hydrodynamic diameter and zeta potential of nanostructured lipid carriers: Emphasizing some parameters for correct measurements

Particle size and zeta potential are basic information about nanoparticles intended for biomedical applications and other of scientific interest. Modern techniques for measuring these properties have become popular due to their simplicity and ease of use, but unfortunately, this has led to an inadequate understanding of the fundamental operational principles and physical meaning of the measured parameters. Consequently, the quality and interpretation of the reported data and the possibility to make comparisons between different studies are being questioned. As a contribution to this respect, this study stresses the importance of suitable experimental conditions to estimate the hydrodynamic diameter and the surface charge of lipid nanoparticles, specifically in nanostructured lipid carriers (NLC) stabilized by nonionic surfactants, by using dynamic light scattering and laser Doppler electrophoresis, respectively, to provide reliable information.

An Overview of Second Generation Nanoparticles− Nanostructure Lipid Carrier Drug Delivery System

Lipids are used as vehicles for the preparation of various formulations prescribed for administrations, including emulsions, ointments, suspension, tablets, and suppositories. The first parental nano-emulsion was discovered from the 1950s when it was added to the intravenous administration of lipid and lipid-soluble substances. Lipid-based drug delivery systems are important nowadays. Solid nanoparticles (SLN) and Nanostructured lipid carriers (NLC) are very proficient due to the ease of production process, scale-up capability, bio-compatibility, the biodegradability of formulation components and other specific features of the proposed route. The administration or nature of the materials must be loaded into these delivery systems. The main objectives of this review are to discuss an overview of second-generation nanoparticles, their limitations, structures, and route of administration, with emphasis on the effectiveness of such formulations. NLC is the second generation of lipid nanoparticles having a structure like nanoemulsion. The first generation of nanoparticles was SLN. The difference between both of them is at its core. Both of them are a colloidal carrier in submicron size in the range of 40-1000 nm. NLC is the most promising novel drug delivery system over the SLN due to solving the problem of drug loading and drug crystallinity. Solid and liquid lipids combination in NLC formation, improve its quality as compare to SLN. NLC has three types of structures: random, amorphous, and multiple. The random structure containing solid-liquid lipids and consisting crystal and the liquid lipid irregular in shape; thereby enhance the ability of the lipid layer to pass through the membrane. The second is the amorphous structure. It is less crystalline in nature and can prevent the leakage of the loaded drug. The third type is multiple structures, which have higher liquid lipid concentrations than other types. The excipients used to form the NLC are bio-compatible, biodegradable and non-irritating, most of which can be detected using GRAS. NLC is a promising delivery system to deliver the drug through pulmonary, ocular, CNS, and oral route of administration. Various methods of preparation and composition of NLC influence its stability Parameters. In recent years at the educational level, the potential of NLC as a delivery mechanism targeting various organs has been investigated in detail.

Nanostructured lipid carrier-based smart gel: a delivery platform for intra-articular therapeutics

The promising potential of nano-structured lipid carrier (NLC) polymeric gel of CUR as an effective treatment for rheumatoid arthritis by intra-articular route of administration was investigated. NLC composed of cetylpalmitate, Labrafac PG & Captex 200, Tween 80 and Labrasol. The hot homogenization method employed by melt ultrasonication was used. The formulated NLC dispersions were characterized and were suitably dispersed into the matrix of pluronic F-127(PLF-127) and pluronic F-68 (PLF-68). A two-factor three-level full factorial design was employed to deduce the optimal concentrations of PLF-127 and PLF-68. The optimized formulations were sterilized by gamma radiation. The formulated NLC smart gels were characterized and evaluated for various parameters. The efficacy evaluation by antigen-induced monoarthritis model and biocompatibility testing by histopathological studies was performed. Formulated NLCs exhibited an average particle size of 165.12 nm, entrapment efficiency of 72.15%, and zeta potential of −21.67 mV. The optimized CUR-NLC smart gel was demonstrated to have a sol–gel transformation at 33.21 °C and 94.32% drug release at 84 h. NLC’s which were sterile and easily syringeable, continued to remain within the colloidal range. CUR-NLC smart gels were found to be biocompatible and showed a significant reduction in rat knee joint inflammation compared to free drug.

Lidocaine loaded nanostructured lipid carriers for prolonged local anesthesia: in vitro and in vivo studies

Lidocaine is widely used to manage local pain. The marketed dosage forms including gels, injections and ointments showed short duration of action (< 2 h). In this paper, the efforts have being made to develop lidocaine loaded nanostructured lipid carrier (NLCs), which on penetration in the skin layer showed prolong local anesthesia. The NLCs was formulated using gelucire (solid-lipid), capryol (liquid-lipid) and Tween 80. The scanning electron microscopy showed smooth and spherical NLCs. The particle size data indicate the size in the range of 98-221 nm. The ex vivo permeation data of lidocaine from the NLCs showed sustain release up to 10 h in comparison to the marketed ointment (up to 4 h). The high lidocaine retention in the skin tissue was observed with NLCs batches. The NLCs was safe in the skin irritation study. The radiant heat tail flick and sciatic nerve block model showed prolong local anesthesia in comparison to the marketed ointment. The study demonstrate the potential of nanostructured lipid carriers for prolong the local anesthesia for the effective management of local pain.

β-Sitosterol Loaded Nanostructured Lipid Carrier: Physical and Oxidative Stability, In Vitro Simulated Digestion and Hypocholesterolemic Activity.

The objective of the present study was to explore the potential of nanostructured lipid carriers (NLCs) for improving the oral delivery of β-sitosterol, a poorly water-soluble bioactive component with hypocholesterolemic activity. Two β-sitosterol formulations with different solid lipid compositions were prepared by melt emulsification, followed by the sonication technique, and the effect of storage conditions and simulated digestion on the physical, chemical and oxidative stability, bioaccessibility and release were extensively studied. Both NLC preparations remained relatively stable during the four weeks of storage at different conditions (4, 25 and 40 °C), with more superior stability at lower temperatures. The in vitro digestion experiment indicated a high physical stability after exposure to the simulated mouth and stomach stages and an improved overall β-sitosterol bioaccessibility at the end of the digestion. The NLCs presented an increased solubility and gradual release which could be justified by the remarkable affinity of β-sitosterol to the complex lipid mixture. An in vivo study demonstrated an improved reduction in the total cholesterol and low-density lipoprotein cholesterol plasma levels in mice compared with the drug suspension. These investigations evidenced the potential of the developed NLC formulations for the enhancement of solubility and in vivo performance of β-sitosterol.

Enzyme-Loaded Gel Core Nanostructured Lipid Carriers to Improve Treatment of Lysosomal Storage Diseases: Formulation and In Vitro Cellular Studies of Elosulfase Alfa-Loaded Systems.

Mucopolysaccharidosis IVA (Morquio A) is a rare inherited metabolic disease caused by deficiency of the lysosomal enzyme N-acetylgalatosamine-6-sulfate-sulfatase (GALNS). Until now, treatments employed included hematopoietic stem cell transplantation and enzyme replacement therapy (ERT); the latter being the most commonly used to treat mucopolysaccharidoses, but with serious disadvantages due to rapid degradation and clearance. The purpose of this study was to develop and evaluate the potential of nanostructured lipid carriers (NLCs) by encapsulating elosulfase alfa and preserving its enzyme activity, leading to enhancement of its biological effect in chondrocyte cells. A pegylated elosulfase alfa-loaded NLC was characterized in terms of size, ζ potential, structural lipid composition (DSC and XRD), morphology (TEM microscopy), and stability in human plasma. The final formulation was freeze-dried by selecting the appropriate cryoprotective agent. Viability assays confirmed that NLCs were non-cytotoxic to human fibroblasts. Imaging techniques (confocal and TEM) were used to assess the cellular uptake of NLCs loaded with elosulfase alfa. This study provides evidence that the encapsulated drug exhibits enzyme activity inside the cells. Overall, this study provides a new approach regarding NLCs as a promising delivery system for the encapsulation of elosulfase alfa or other enzymes and the preservation of its activity and stability to be used in enzymatic replacement therapy (ERT).

Solidified nanostructured lipid carrier (S-NLC) for enhancing the oral bioavailability of ezetimibe

Current study explores the potential of nanostructured lipid carriers (NLCs) for bioavailability enhancement of ezetimibe on oral administration. Microemulsion technique was employed for the development of NLCs. Along with NLC, a nanoemulsion of ezetimibe was also prepared and compared with optimized NLC for morphology and in vitro release. An aqueous dispersion of the optimized NLC was transformed into a stable free-flowing solidified-NLC (S-NLC) by adsorbing it on Neusilin® US2. The S-NLC was evaluated for differential scanning calorimetry (DSC), X-ray diffraction (XRD), scanning electron microscopy (SEM), drug content, dissolution profile, pharmacodynamics and pharmacokinetics in rats. DSC and XRD studies showed reduced crystallinity of ezetimibe in the form of S-NLC. SEM study supports these results, as no drug precipitation was observed on the carrier surface. Dissolution study exhibited a notable rise in the dissolution of the drug from S-NLC as compared to pure drug and marketed tablet. In vivo evaluation in rats revealed a significant reduction in the level of total cholesterol as compared to positive control and drug suspension group, whereas for high-density lipoprotein, non-significant change was observed. Pharmacokinetics study showed a significant rise in the bioavailability of ezetimibe using NLC formulation as compared to the suspension and marketed tablet.

Fabrication and Characterization of Pluronic F68 and Phospholipon 90g Embedded Nanoformulation for Sertraline Delivery: An Optimized Factorial Design Approach and In Vivo Study

Objective: The objective of present work was to utilize the potential of nanostructured lipid carriers (NLCs) form improvement in bioavailability of Sertraline as antidepressant drug formulated by emulsification- solvent evaporation technique with some modification. NLC is the blend of solid lipid, liquid lipid and surfactant for encapsulation of poor water soluble actives. Design: A full 32 factorial design was utilized to study the effect of two independent parameters namely solid lipid to liquid lipid concentration and stabilizer concentration on the entrapment efficiency of the prepared NLCs. The sertraline NLC formulation was characterized with respect to particle size, polydispersity index (PDI), zeta-potential, encapsulation efficiency and physical morphology. Result: The NLC formulation had an average diameter of 96.59 nm, PDI of 0.192, zeta-potential of -39.88 mV, and encapsulation efficiency of 97%, respectively. Conclusion: The NLC formulation for sertraline encapsulation has been successfully developed and is suitable for nose to brain delivery system due to their nano-size and sta

Quercetin Loaded Nanostructured Lipid Carriers for Nose to Brain Delivery: In Vitro and In Vivo Studies

The main objective of the study was to utilize potential of nanostructured lipid carriers of the quercetin for direct nose to brain delivery of drug as tool for the targeted delivery. The aim of this study was to prepare and characterize quercetin loaded NLC and to study its brain distribution. Novel QUE-NLCs were formulated. NLC Formulation was evaluated for various physicochemical properties such as particle size, zeta potential, drug loading, percent entrapment efficiency, morphology, in vitro drug release, and histopathology. The size of the QUE-NLC was about 118.2nm with poly dispersity index of 0.220 and zeta potential of -20.1 mV. Nose to brain distribution studies were performed using wistar rats.QUE-NLC exhibited sustained delivery of drug. Significant targeting to brain was achieved when compare to quercetin.The result showed that NLCs might be the promising approach for the nose to brain delivery of quercetin.

Role of surface capping on the cytotoxicity of silica nanoparticles in rat alveolar epithelial cells

The aim of the present work was to develop and evaluate the potential of nanostructured lipid carriers associated with caffeine extracted from Coffee Silverskin (NLC-CS), a food by-product, as a new possible topical therapy of cellulitis. Caffeine gain increasing research interest due to their cosmetic potential, particularly in gynoid lipodystrophy, commonly known as cellulite. NLC-CS were prepared via double emulsion technique using polysorbate 60 as surfactant and characterized for their morphology, particle size, zeta potential, association encapsulation and stability. The in vitro skin permeation studies were performed on Franz diffusion cells using pig skin ear as permeation membrane and the optimized formulation was compared with a hydroalcoholic solution of Coffee silverskin (CS) extract.NLC-CS were within the nanosized range (≈200 nm), with a low polydispersity index (<0.25) and zeta potential values around −30 Mv, presenting storage stability up to 180 days at 25 °C/65% relative humidity (RH) and 40 °C/75% RH. The association efficiency (AE) of caffeine was about 30% at production time and after storage period. Cryo-SEM images confirmed the spherical shape of NLC-CS.The in vitro skin permeation study demonstrated that NLC-CS had a similar skin permeation profile when compared to caffeine in CS extract.

Nose to Brain Delivery of Rivastigmine by In Situ Gelling Cationic Nanostructured Lipid Carriers: Enhanced Brain Distribution and Pharmacodynamics

Present investigation explores the potential of nanostructured lipid carriers (NLCs) for nose to brain delivery of rivastigmine (RV), which is further enhanced by incorporating into an in situ gelling system, increasing retention in nasal cavity. NLCs having particle size of 123.2 ± 2.3 nm, entrapment efficiency of 68.3 ± 3.4%, and zeta potential of 32 ± 1.2 mV was fabricated by a scalable method. Pharmacokinetics showed sustained release of intranasal (IN) and intravenous (IV) NLCs compared with RV solution by same route, with significantly higher AUC and Thalf. Biodistribution indicated blood brain barrier penetrating potential of IV NLCs (457.24 ± 38.41.12 ng/mL) and IN NLCs (736.42 ± 34.54 ng/mL) with 4.6 and 5.3-fold enhancement in brain concentrations compared with IV and IN solution of RV. Similar results reflected in pharmacodynamics, indicating faster regain of memory loss in amnesic mice with 5-fold decrease in escape latency with NLCs compared with plain RV solution by IV and IN routes respectively. Sub-acute toxicity studies demonstrated safety of developed formulations to vital organs without any hematological and biochemical changes compared with control group. Moreover, nasal toxicity studies of NLCs showed no signs of inflammation, maintaining the integrity of ciliary epithelial cells, thus confirming safety of the formulation for its intended nasal application.

Pharmacokinetic studies and anticancer activity of curcumin-loaded nanostructured lipid carriers

In order to investigate the potential of nanostructured lipid carriers for efficient and targeted delivery of curcumin, the pharmacokinetic parameters of curcumin-loaded nanostructured lipid carriers (Cur-NLC) were evaluated in rats after a single intraperitoneal dose of Cur-NLC. In addition, the anticancer activity of Cur-NLC against human lung adenocarcinoma A549 cells was verified by a cellular uptake study, and a cytotoxicity and apoptosis assay. Bioavailability of Cur-NLC was better than that of native curcumin (p > 0.01), as seen from the area under the plasma concentration-time curve (AUC), maximum plasma concentration (Cmax), mean residence time (MRT) and total plasma clearance (CLz/F). Cur-NLC has a more obvious lung-targeting property in comparison with native curcumin. Cur-NLC showed higher anticancer activity in vitro against A549 cells than native curcumin (IC50 value of 5.66 vs. 9.81 mg L-1, respectively). Meanwhile, Cur-NLC treated A549 cells showed a higher apoptosis rate compared to that of native curcumin. These results indicate that NLC is a promising system for the delivery of curcumin in the treatment of lung adenocarcinoma.

Fabrication, optimization and characterization of Triamcinolone acetonide loaded nanostructured lipid carriers for topical treatment of psoriasis: Application of Box Behnken design, in vitro and ex vivo studies

Psoriasis is a highly inflamed, chronic, autoimmune skin disorder affecting 2–5% of the world population. Complete cure for psoriasis is still lacking and there remains a substantial challenge for world health systems to explore a new drug moiety or delivery system which could safely and effectively manage psoriasis without compromising patient compliance. Present work was aimed to develop, optimize and investigate the potential of nanostructured lipid carriers (NLC) for secure and efficient delivery of Triamcinolone acetonide (TA). TA loaded NLCs were effectively fabricated by modified microemulsion method and examined for particle size, zeta potential, polydispersity index, drug entrapment efficiency, drug loading, transmission electron microscopy, X ray diffraction and Differential scanning calorimetry study. Release study demonstrated prolonged TA release from NLCs TA suspension following Higuchi release kinetics with r2 = 0.995, while plain TA suspension showed quicker drug release obeying Zero order kinetics with R2 value of 0.993. In vitro skin distribution studies demonstrated the presence of significant quantity of TA on the epidermis when treated with NLCs TA suspension. Adverse side effects linked with systemic exposure might be removed by selective drug accumulation in the epidermis. Conclusively TA loaded NLCs might be a efficient carrier for effective management of psoriasis.

Nanostructured lipid carriers of olmesartan medoxomil with enhanced oral bioavailability

The current study explores the potential of nanostructured lipid carriers (NLCs) for oral bioavailability enhancement of olmesartan medoxomil (OLM) by systemic design approach. OLM-NLC was successfully prepared with optimized process parameters (i.e. amount of liquid lipid, total amount of lipid, drug content and surfactant concentration) using the Box-Behnken design of experiments for different response parameters (i.e. particle size, Polydispersity index and entrapment efficiency). Further, optimized formulation was validated which depicted nano size, homogenous distribution with optimum entrapment efficiency. OLM-NLC was characterized by different techniques viz. differential scanning calorimetry (DSC), powder X-Ray diffraction (PXRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM), which showed reduced crystallinity of the drug with smooth spherical appearance of nanoparticles. Formulation was found to be stable in simulated gastric fluids as no significant changes were found in size, PDI and entrapment efficiency. In vitro release showed extended release of OLM from OLM-NLC. In vitro cellular uptake study revealed 5.2 folds higher uptake of nanoparticles as compare to the free drug, when incubated with Caco-2 cells. In vivo performance showed that AUCtotal and Cmax of OLM-NLC were found significantly (P<0.01) higher as compare to the free drug. Overall, the present study successfully reports the improvement of oral bioavailability of olmesartan medoxomil.

Preparation and Characterisation of Nobiletin-Loaded Nanostructured Lipid Carriers

The objective of this manuscript was to investigate and optimise the potential of nanostructured lipid carriers (NLCs) as a carrier system for nobiletin (NOB), which was prepared by high-pressure homogenisation method. Additionally, this study was focused on the application of NOB-loaded NLC (NOB-NLC) in functional food. Response surface method with a three-level Box–Behnken design was validated through analysis of variance, and the robustness of the design was confirmed through the correspondence between the values measured in the experiments and the predicted ones. Properties of the prepared NOB-NLC, such as Z-average, polydispersity, entrapment efficiency, zeta potential, morphology, and crystallinity, were investigated. NOB-NLC exhibited a spherical shape with a diameter of 112.27 ± 5.33 nm, zeta potential of −35.1 ± 2.94 mV, a polydispersity index of 0.251 ± 0.058, and an EE of 81.06% ± 6.02%. Results from X-ray diffraction and differential scanning calorimetry of NOB-NLC reviewed that the NOB crystal might be converted to an amorphous state. Fourier transform infrared spectroscopic analysis demonstrated that chemical interaction was absent between the compound and lipid mixture in NOB-NLC.