Nanostructured Lipid Carrier

This study presents the formulation of nanostructured lipid carrier encapsulated Zingiber
zerumbet oil (NLC-ZZ) using ultrasonication technique. NLC is the blend of solid lipid, liquid lipid
and surfactant for encapsulation of poor water soluble actives. The NLC-ZZ formulation was
characterized with respect to particle size, polydispersity index (PDI), zeta-potential, encapsulation
efficiency and physical morphology. The NLC-ZZ formulation had an average diameter of 96.59 nm,
PDI of 0.192, zeta-potential of -39.88 mV, and encapsulation efficiency of 90%, respectively. The
NLC formulation for Zingiber zerumbet oil encapsulation has successfully developed and suitable for
transdermal delivery system due to their nano-size and stability.

A nanostructured lipid carrier (NLC) loaded with doxorubicin (DOX) has been shown to be cytotoxic against the human cancer cell lines A549 and MCF-7/Adr. In attempts to improve formulation characteristics, enhance pharmacokinetics and antitumor effects, we modified the surface of these NLC with an alternating layer-by-layer (LbL) assembly of polycation and polyanion polyelectrolytes and an additional coating with PEG using a simple method of core shell attachment. The formulation had a narrow size distribution, longer residence in the blood, lower accumulation in the liver, higher
accumulation in tumors and a significant tumor growth inhibition effect. Thus, NLC–DOX nanopreparations complexes modified by LbL coating have the potential to enhance the anticancer effects of
DOX against tumors.

The prevalence of non-communicable diseases (NCDs) is showing an increasing trend in most developed and developing nations. For diseases such as Type 2 diabetes mellitus and hypertension, the prevalence is already at a crisis level. Despite intensive efforts and billions of dollars spent in terms of research, health education and preventive strategies, there is still no solution in sight as the number of deaths attributable to the NCDs continues to rise. On the research front, there is still a lot more to be done and arguably the best approach would be setting up 'population laboratories' and studying individuals over time i.e. a cohort design. The Malaysian Cohort is a one of the latest cohorts amongst the developing nations. But having a population with 3 major ethnic groups gives the cohort an added advantage in terms of comparative analysis across ethnic groups. At recruitment written informed consent was taken and each participant went through a questionnaire-based interview, a biophysical examination, and gave blood and urine for baseline tests and also storage. The project has recruited a total 106,527 participants aged 35 years and above from the year 2007 to 2012. 53% were females and 47% males. A total of 71.4% or the participants are from the urban areas whilst 28.6% are from the rural communities. The prevalence of Type 2 diabetes mellitus, amongst the Cohort population is 16.6%, hypertension at 46.5%, obesity at 17.7% and hypercholesterolemia (>5.2 mmol/L) at 75.7%. The percentage of those with impaired fasting glucose is 10.4%. Clearly the trends are increasing when compared to the data from the National Health Morbidity Survey by the Ministry of Health Malaysia where in 2006, the prevalence of diabetes was at 14.6%. Clearly, the lack of physical activity and also poor diet practices have caused this surge in numbers of those with NCDs. There are already 1131 deaths so far and 226 are due to cancers. The majority of those who died of cancers were asymptomatic when they were recruited hence giving the opportunity to analyse the blood specimens for biomarkers for early detection. There is a lot of analysis which can be done on the baseline data to identify risk factors. The follow-up phase has just started and more than 2500 participants have been revisited so far and re-interviewed, re-measured plus also having another set of biospecimens taken. An intervention study on those with impaired fasting glucose is also being planned. In summary, the Malaysian Cohort study confirms the statistics which we already known but the data and biospecimens will provide endless possibilities in terms of research especially in terms of looking at gene-environment interaction and also comparison across ethnic groups. Identification of risk factors, whether lifestyle or genetic in origin, will be crucial in terms of applying the personalised, preventive and participatory approaches. Like any cohort study, the value of the Malaysian Cohort will certainly increase with time.

The prevalence of non-communicable diseases (NCDs) is showing an increasing trend in most developed and developing nations. For diseases such as Type 2 diabetes mellitus and hypertension, the prevalence is already at a crisis level. Despite intensive efforts and billions of dollars spent in terms of research, health education and preventive strategies, there is still no solution in sight as the number of deaths attributable to the NCDs continues to rise. On the research front, there is still a lot more to be done and arguably the best approach would be setting up 'population laboratories' and studying individuals over time i.e. a cohort design. The Malaysian Cohort is a one of the latest cohorts amongst the developing nations. But having a population with 3 major ethnic groups gives the cohort an added advantage in terms of comparative analysis across ethnic groups. At recruitment written informed consent was taken and each participant went through a questionnaire-based interview, a biophysical examination, and gave blood and urine for baseline tests and also storage. The project has recruited a total 106,527 participants aged 35 years and above from the year 2007 to 2012. 53% were females and 47% males. A total of 71.4% or the participants are from the urban areas whilst 28.6% are from the rural communities. The prevalence of Type 2 diabetes mellitus, amongst the Cohort population is 16.6%, hypertension at 46.5%, obesity at 17.7% and hypercholesterolemia (>5.2 mmol/L) at 75.7%. The percentage of those with impaired fasting glucose is 10.4%. Clearly the trends are increasing when compared to the data from the National Health Morbidity Survey by the Ministry of Health Malaysia where in 2006, the prevalence of diabetes was at 14.6%. Clearly, the lack of physical activity and also poor diet practices have caused this surge in numbers of those with NCDs. There are already 1131 deaths so far and 226 are due to cancers. The majority of those who died of cancers were asymptomatic when they were recruited hence giving the opportunity to analyse the blood specimens for biomarkers for early detection. There is a lot of analysis which can be done on the baseline data to identify risk factors. The follow-up phase has just started and more than 2500 participants have been revisited so far and re-interviewed, re-measured plus also having another set of biospecimens taken. An intervention study on those with impaired fasting glucose is also being planned. In summary, the Malaysian Cohort study confirms the statistics which we already known but the data and biospecimens will provide endless possibilities in terms of research especially in terms of looking at gene-environment interaction and also comparison across ethnic groups. Identification of risk factors, whether lifestyle or genetic in origin, will be crucial in terms of applying the personalised, preventive and participatory approaches. Like any cohort study, the value of the Malaysian Cohort will certainly increase with time.

Solid lipid nanoparticles (SLN) and nano- structured lipid carriers (NLC) have been intensively investigated for different applications, including their use as drug and gene delivery systems. Different techniques have been employed to produce lipid nanoparticles, of which high pressure homogenization is the standard technique that is adopted nowadays. Although this method has a high efficiency, does not require the use of organic solvents, and allows large- scale production, some limitations impede its applica- tion at laboratory scale: the equipment is expensive, there is a need of huge amounts of surfactants and co- surfactants during the preparation, and the operating conditions are energy intensive. Here, we present the microemulsion extrusion technique as an alternative method to prepare lipid nanoparticles. The parameters to produce lipid nanoparticles using microemulsion extru- sion were established, and the lipid particles produced (SLN, NLC, and liposomes) were characterized with regard to size (from 130 to 190 nm), zeta potential, and drug (mitoxantrone) and gene (pDNA) delivery prop- erties. In addition, the particles’ in vitro co-delivery capacity (to carry mitoxantrone plus pDNA encoding the phosphatase and tensin homologue, PTEN) was tested in normal (BALB 3T3 fibroblast) and cancer (PC3 prostate and MCF-7 breast) cell lines. The results show that the microemulsion extrusion technique is fast, inexpensive, reproducible, free of organic solvents, and suitable for small volume preparations of lipid nano- particles. Its application is particularly interesting when using rare and/or costly drugs or ingredients (e.g., cationic lipids for gene delivery or labeled lipids for nanoparticle tracking/diagnosis).

The present research aimed at developing an injection-free nanoparticulated formulation in multiple emulsion
form, for oral delivery of insulin, which otherwise undergoes degradation in the gastric environment if administered
orally. Insulin-polymeric nanoparticles were prepared using layer by layer (LbL) adsorption method and incorporated into
an emulsion to form a nanoparticulated multiple emulsion. Using 0.6 M sodium chloride, the insulin nanoaggregates of
300-400 nm size were obtained about a yield of 94%. The characteristics of a representative nanoparticle were as follows: particle size - 391.9±0.41 nm, polydispersity index -0.425, zeta potential- +20.6 mv, encapsulation efficiency- 86.7±1.42%
and percentage entrapment efficiency of the insulin-polymeric nanoparticles in the inner aqueous phase of emulsion was
84.6 %. The FT-IR analysis confirms that there were no drug interactions with the polymers. Stability analysis carried out
for 3 months at 8-40 °C, showed only minor changes at the end period. The release kinetics of the nanoparticulated multiple
emulsion at pH 7.4 followed first order kinetics and obeyed the Fickian law. However, at pH 2.0 the release kinetics
from nanoparticulated multiple emulsion followed zero order kinetics without obeying to the Fickian law. In conclusion,
our data demonstrate that the nanoparticulated multiple emulsion formulation has good release characteristics and imparted a tolerable protection for insulin at different pH conditions, which may be exploited for oral administration.

Nanoparticles are one part of nanotechnology that is widely developed on Nano medicine and Nanodermatology. Nanomaterial is a system that has sizes ranging from 1-1000 nm (or 10-9-10-7 m). Besides being used for drug delivery, nanoparticles are now also widely used in cosmetic preparations to enhance the performance of active ingredients, protect active ingredients from degradation, alert local action, and prevent active ingredients from entering the bloodstream. Nanoparticles used in cosmetic dosage formulations include Nano-emulsion, Nano-crystal, micelles, polymeric Nano-capsules, noisome, liposomes, nanostructured lipid carriers, solid lipid nanoparticles. Nano Lipid Carrier (NLC) which is the second generation of Solid Lipid Nano particle (SLN) delivery system. NLC consists of a certain amount of lipid matrix and liquid lipids, providing enough space to accommodate drug molecules and consequently enhancing drug trapping capacity, preventing drug release, improving drug stability and increasing flexibility to modulate drug release.

Paclitaxel (PTX) is an anticancer drug having poor aqueous solubility and low bioavailability. Formulation of PTX into Nanostructure lipid carriers (NLC) could be a potential way to enhance PTX aqueous solubility and bioavailability hence increases efficacy and decreases side effects. Eight PTX-NLC formulae were prepared using homogenization-ultrasonication technique. Characterization of the nanoparticles was done by transmission electron microscopy and by measurement of particle size, poly dispersibility index and zeta potential. Encapsulation efficiency, drug loading, and In Vitro release were measured. Particle size ranged between 172.8 ± 0.8 to 378.2 ± 1.8 nm and zeta potential between-18.6 ± 0.4 to-28.1 ± 1.2 mV. High EE and DL were obtained due to incorporation of liquid lipid and the In Vitro release showed prolonged time dependent release compared to Taxol ®. NLC-3 had the best results among the eight prepared formulae. In Vitro cytotoxicity of NLC-3 was evaluated on MCF-7 cell line and compared to pure PTX powder and Taxol ®. These findings show that NLC is a potential carrier to improve efficacy and enhance PTX delivery.

The prevalence of non-communicable diseases (NCDs) is showing an increasing trend in most developed and developing nations. For diseases such as Type 2 diabetes mellitus and hypertension, the prevalence is already at a crisis level. Despite intensive efforts and billions of dollars spent in terms of research, health education and preventive strategies, there is still no solution in sight as the number of deaths attributable to the NCDs continues to rise. On the research front, there is still a lot more to be done and arguably the best approach would be setting up 'population laboratories' and studying individuals over time i.e. a cohort design. The Malaysian Cohort is a one of the latest cohorts amongst the developing nations. But having a population with 3 major ethnic groups gives the cohort an added advantage in terms of comparative analysis across ethnic groups. At recruitment written informed consent was taken and each participant went through a questionnaire-based interview, a biophysical examination, and gave blood and urine for baseline tests and also storage. The project has recruited a total 106,527 participants aged 35 years and above from the year 2007 to 2012. 53% were females and 47% males. A total of 71.4% or the participants are from the urban areas whilst 28.6% are from the rural communities. The prevalence of Type 2 diabetes mellitus, amongst the Cohort population is 16.6%, hypertension at 46.5%, obesity at 17.7% and hypercholesterolemia (>5.2 mmol/L) at 75.7%. The percentage of those with impaired fasting glucose is 10.4%. Clearly the trends are increasing when compared to the data from the National Health Morbidity Survey by the Ministry of Health Malaysia where in 2006, the prevalence of diabetes was at 14.6%. Clearly, the lack of physical activity and also poor diet practices have caused this surge in numbers of those with NCDs. There are already 1131 deaths so far and 226 are due to cancers. The majority of those who died of cancers were asymptomatic when they were recruited hence giving the opportunity to analyse the blood specimens for biomarkers for early detection. There is a lot of analysis which can be done on the baseline data to identify risk factors. The follow-up phase has just started and more than 2500 participants have been revisited so far and re-interviewed, re-measured plus also having another set of biospecimens taken. An intervention study on those with impaired fasting glucose is also being planned. In summary, the Malaysian Cohort study confirms the statistics which we already known but the data and biospecimens will provide endless possibilities in terms of research especially in terms of looking at gene-environment interaction and also comparison across ethnic groups. Identification of risk factors, whether lifestyle or genetic in origin, will be crucial in terms of applying the personalised, preventive and participatory approaches. Like any cohort study, the value of the Malaysian Cohort will certainly increase with time.

Paclitaxel (PTX) is an anticancer drug having poor aqueous solubility and low bioavailability. Formulation of PTX into Nanostructure lipid carriers (NLC) could be a potential way to enhance PTX aqueous solubility and bioavailability hence increases efficacy and decreases side effects. Eight PTX-NLC formulae were prepared using homogenization-ultrasonication technique. Characterization of the nanoparticles was done by transmission electron microscopy and by measurement of particle size, poly dispersibility index and zeta potential. Encapsulation efficiency, drug loading, and In Vitro release were measured. Particle size ranged between 172.8 ± 0.8 to 378.2 ± 1.8 nm and zeta potential between-18.6 ± 0.4 to-28.1 ± 1.2 mV. High EE and DL were obtained due to incorporation of liquid lipid and the In Vitro release showed prolonged time dependent release compared to Taxol ®. NLC-3 had the best results among the eight prepared formulae. In Vitro cytotoxicity of NLC-3 was evaluated on MCF-7 cell line and compared to pure PTX powder and Taxol ®. These findings show that NLC is a potential carrier to improve efficacy and enhance PTX delivery.

Nanotechnology can produce materials with unique features compared to their bulk counterparts, which can be useful for medical applications (i.e. nanomedicine). Among the therapeutic agents used in nanomedicine, small molecules or biomacromolecules, such as proteins or genetic materials, can be designed for disease diagnostics and treatment. To transport these biomacromolecules to the target cells, nanomedicine requires nanocarriers. Solid lipid nanoparticles (SLNs) are among the promising nanocarriers available, because they can be made from biocompatible materials and present high stability (over one year). In addition, upon the binding genetic material, SLNs form SLNplexes. However, little is yet known about how cells process these SLNplexes-in particular, how internalization and endosome acidification affects the transfection mediated by SLNplexes. Therefore, we aim to investigate how these processes affect SLNplex transfection in HEK293T cells. We find that the SLNplex is mainly internalized by clathrin-mediated endocytosis, which is a fast and reliable pathway to transfection, leading to approximately 60% transfection efficiency. Interestingly, upon acidification (below pH 5.0), the SLN seems to release its DNA content, which can be an essential step for SLNplex transfection. The underlying mechanisms described in this work may help improve SLNplex formulations and transfection efficiency. Moreover, these advances can improve the field of nanomedical research and bring new ways to cure diseases. Supplementary material for this article is available online

13 14 Aims: the objective of the current study was to formulate nystatin (Nyst) into nanostructured lipid carriers (NLCs) to enhance its antifungal activity. Place and Duration of Study: Department of pharmaceutical technology, national research centre, Egypt, between mars 2011 to april 2013 Methodology: Nyst loaded NLCs (NYST-NLCs) were prepared by the hot homogenization and ultrasonication method followed by evaluation of its topical effect on the cutaneous candidiasis. The prepared Nyst-NLCs were characterized for entrapment efficiency, particle size, zeta potential, morphology (transmission electron microscopy), thermal characterisation (differential scanning calorimetry) and in vitro drug release. The study design involves the investigation of the effect of three independent variables namely liquid lipid type (Miglyol 812 and Squalene), liquid lipid concentration (30 and 50%) and Nyst concentration (0.125 and 0.25%). A stability study for 6 months was performed. A microbiological study was conducted in male rats infected with Candida albicans. Results: NLC dispersions were spherical in shape with particle size ranging from 68.06±6.56 to 141.8±3.33 nm. The entrapment efficiencies ranged from 45.50±2.34 to 92.73±0.33% with zeta potential ranging from-26.2 to-39.2 mV. The stability studies done for 6 months indicated that Nyst-NLCs were stable for more than 6 monthes.the microbiological studies showed a least number of colonies forming units (cfu/ml) were recorded for the selected Nyst-NLCs compared to the drug solution and the Nystatin® cream present in the market. Conclusion: It can be fulfilled from this work that NLCs represent promising carrier for topical delivery of Nyst offering good physical stability, high entrapment efficiency and controlled drug release. Nyst-NLCs are a good candidate for cutaneous treatment of fungal infection Note: Review paper may have different types of subsections. 15

Dacarbazine (Dac) is one of the most commonly used chemotherapy drugs for treating various cancers. However, its poor water solubility, short half-life in blood circulation, low response rate and high side effect limit its application. This study aimed to improve the drug solubility and prolong drug release by developing nanostructured lipid carriers (NLCs) for Dac delivery. The NLC and Dac-encapsulated NLC were synthesized with precirol ATO 5 and isopropyl myristate as lipids, tocopheryl polyethylene glycol succinate, soybean lecithin and Kolliphor P 188 as co-surfactants. The NLCs with controlled size were achieved using high shear dispersion following solidification of oil-in-water emulsion. For Dac encapsulation, the smallest NLC with 155 ± 10 nm in size, 0.2 ± 0.01 polydispersion index and -43.4 ± 2 mV zeta potential was selected. The resultant DLC-Dac possessed size, polydispersion index and zeta potential of 190 ± 10, 0.2 ± 0.01, and -43.5 ± 1.2, respectively. The drug encapsulation efficiency and drug loading were 98.5 % and 14 %, respectively. In vitro drug release study showed a biphasic pattern, with 50 % released in the first 2 h, and the remaining released sustainably for up to 30 h. This is the first report on the development of NLC for Dac delivery, implying that NLC could be a new potential candidate as drug carrier to improve the therapeutic profile of Dac.

Zerumbone, a natural dietary lipophilic compound with low water solubility (1.296 mg/L at 25°C) was used in this investigation. The zerumbone was loaded into nanostructured lipid carriers using a hot, high-pressure homogenization technique. The physicochemical properties of the zerumbone-loaded nanostructured lipid carriers (ZER-NLC) were determined. The ZER-NLC particles had an average size of 52.68 ± 0.1 nm and a polydispersity index of 0.29 ± 0.004 µm. Transmission electron microscopy showed that the particles were spherical in shape. The zeta potential of the ZER-NLC was −25.03 ± 1.24 mV, entrapment efficiency was 99.03%, and drug loading was 7.92%. In vitro drug release of zerumbone from ZER-NLC was 46.7%, and for a pure zerumbone dispersion was 90.5% over 48 hours, following a zero equation. Using the MTT assay in human T-cell acute lymphoblastic leukemia (Jurkat) cells, the half maximal inhibitory concentration (IC 50 ) of ZER-NLC was 5.64 ± 0.38 µg/mL, and for free zerumbone was 5.39 ± 0.43 µg/mL after 72 hours of treatment. This study strongly suggests that ZER-NLC have potential as a sustained-release drug carrier system for the treatment of leukemia.