Structured Lipids Research Articles

In-vitro and In-vivo Correlation [IVIVC] for Nanoparticulate Drug Delivery Systems

Background and Objective: Lipid based nanoformulations have attracted a lot of attention in recent market because of their nano size properties, high degree of biocompatibility and versatility. Various lipid based formulation are available in market like liposomes, niosomes, solid lipid nanoparticles, nano structured lipid carrier for oral, topical, pulmonary or parenteral route based on disease conditions. Discussion: Whenever lipid based formulation is developed then there is a necessity to check its oral bioavailability in human which is very time consuming process and cost of process is also increases. IVIVC is great tool for such a lipid based nanoformulations. IVIVC predictive mathematical model describes the relationship between an in-vitro property of a dosage form and it's in-vivo response. IVIVC can be used in the development of new pharmaceuticals to reduce the number of human studies during the formulation development and ultimately it decreases time which spent during in-vivo study. Conclusion: Thus, the main objective of an IVIVC is to serve as a surrogate for in-vivo bioavailability study and also support biowaivers. This article provides the information about levels of IVIVC, purpose of IVIVC, factors affecting IVIVC, various steps involved in IVIVC development, IVIVC study in lipid based formulation. Keywords: IVIVC, BCS class, liposomes, niosomes, SLN, transferosomes, lipid based formulations, bioavailability.

Structural difference of palm based Medium- and Long-Chain Triacylglycerol (MLCT) further reduces body fat accumulation in DIO C57BL/6J mice when consumed in low fat diet for a mid-term period

Medium-and-Long Chain Triacylglycerol (MLCT) is a type of structured lipid that is made up of medium chain, MCFA (C8-C12) and long chain, LCFA (C16-C22) fatty acid. Studies claimed that consumption of MLCT has the potential in reducing visceral fat accumulation as compared to long chain triacylglycerol, LCT. This is mainly attributed to the rapid metabolism of MCFA as compared to LCFA. Our study was designed to compare the anti-obesity effects of a enzymatically interesterified MLCT (E-MLCT) with physical blend of palm kernel and palm oil (B-PKOPO) having similar fatty acid composition and a commercial MLCT (C-MLCT) made of rapeseed/soybean oil on Diet Induced Obesity (DIO) C57BL/6J mice for a period of four months in low fat, LF (7%) and high fat, HF (30%) diet. The main aim was to determine if the anti-obesity effect of MLCT was contributed solely by its triacylglycerol structure alone or its fatty acid composition or both.Out of the three types of MLCT, mice fed with Low Fat, LF (7%) E-MLCT had significantly (P<0.05) lower body weight gain (by ~30%), body fat accumulation (by ~37%) and hormone leptin level as compared to both the LF B-PKOPO and LF C-MLCT. Histological examination further revealed that dietary intake of E-MLCT inhibited hepatic lipid accumulation. Besides, analysis of serum profile also demonstrated that consumption of E-MLCT was better in regulating blood glucose compared to B-PKOPO and C-MLCT. Nevertheless, both B-PKO-PO and E-MLCT which contained higher level of myristic acid was found to be hypercholesterolemic compared to C-MLCT.In summary, our finding showed that triacylglycerol structure, fatty acid composition and fat dosage play a pivotal role in regulating visceral fat accumulation. Consumption of E-MLCT in low fat diet led to a significantly lesser body fat accumulation. It was postulated that the MLM/MLL/LMM/MML/LLM types of triacylglycerol and C8-C12 medium chain fatty acids were the main factors that contributed to the visceral fat suppressing effect of MLCT. Despite being able to reduce body fat, the so called healthful functional oil E-MLCT when taken in high amount do resulted in fat accumulation. In summary, E-MLCT when taken in moderation can be used to manage obesity issue. However, consumption of E-MLCT may lead to higher total cholesterol and LDL level.

Design, Development and Invivo Pharmacokinetic Evaluation of Cardiovascular Drug Loaded Nano Structured Lipid Carrier System

The Present investigation is to design, develop and optimize the Nanostructured Lipid Carrier (NLC) containing cardiovascular drug by various independent variables like formulation variables (concentration of phospholipids, surfactants, co-surfactant) and process variables (homogenization time and ultrasonication time). The objective of this research is to choose the suitable and reproducible technique for the formulation of NLC based on dependent variables like Particle Size (PS) in nm, Polydispersity Index (PDI), Zeta potential (ZP) in mV and Entrapment efficiency (EE%) by applying Box Behnken design utilizing - Multiple linear regression method (Design expert 9 software, stat-ease, Inc. USA) and another objective is to prove the enhancement of bioavailability of drug in developed NLC. Comparative invivo pharmacokinetic studies was carried out to determine AUC, Cmax, Tmax , Ke, Vd between drug loaded NLC and commercially available dosage form to prove the enhancement of bioavailability in NLC. From the results obtained, it was concluded that hot homogenization was an optimized technique for the preparation of NLC containing carvedilol and NLC shows better bioavailability when compare to commercial formulation. This data was related to the research project sanctioned by Department of science and technology (DST) entitled “Development of Lipid Nanoparticles Enriched Hydrogels for Transdermal Drug Delivery: Formulation, In vitro Characterization and Pharmacodynamic studies in Animals”

Cs2.5H0.5PW12O40 Encapsulated in Metal–Organic Framework UiO-66 as Heterogeneous Catalysts for Acidolysis of Soybean Oil

The Keggin-type Cs2.5H0.5PW12O40 was encapsulated in metal–organic framework UiO-66 by a simple solvothermal synthesis method. The characterization results showed that the morphology and crystal structure of UiO-66 were preserved, and the heteropolyanion structure remained unchanged after Cs2.5H0.5PW12O40 encapsulation. The prepared Cs2.5H0.5PW12O40@UiO-66 material has demonstrated to be an environmentally benign catalyst in the acidolysis of soybean oil with medium-chain fatty acids for the production of low-calorie structured lipids. The FT-IR CO adsorption experiments revealed that additional Lewis acid sites were formed in the solid acid catalyst, which were essential to catalyze the acidolysis reaction. The solid catalyst could be reused for several recycles without obvious deactivation due to hardly leaching of active components into the reaction mixture.

In Vitro Evidence of Anti-Inflammatory and Anti-Obesity Effects of Medium-Chain Fatty Acid-Diacylglycerols.

Dietary approaches using structured lipids, including medium-chain fatty acids and diacylglycerols, have been adopted for the prevention of obesity-induced chronic inflammation. In an extension to previous studies, medium-chain fatty acid-diacylglycerol enriched dietary oil (MCDG) was prepared by interesterification of canola oil and mediumchain fatty acid-triacylglycerols. The consequent MCDG product was applied to RAW264.7 macrophages followed by the assessment of multiple inflammatory responses. Compared with conventionally used canola and olive oil controls, MCDG suppressed macrophage phagocytosis, as assessed by the uptake of microsphere beads. Furthermore, the production of IL-6 and TNF-α, transcription of COX-2 and iNOS, and expression of CD80 on cell surfaces were downregulated by MCDG in LPS-stimulated macrophages. Subsequently, differentiated 3T3-L1 adipocytes were evaluated for proinflammatory cytokine production and lipid accumulation. IL-6 production was marginally affected and lipid accumulation was inhibited by MCDG. Taken together, these results suggest that MCDG has potential as an alternative oil for cooking in order to prevent obesity-induced inflammation.

Biotechnological and Novel Approaches for Designing Structured Lipids Intended for Infant Nutrition

AbstractHuman milk fat (HMF) is a perfect nutritional source that includes all the required ingredients which are necessary for the growth of infants up to 6 months. Although its composition may differ among mothers or during lactation stage, its unique triacylglycerol (TAG) structure remains constant which is characterized by the presence of palmitic acid (PA) at the sn‐2 position. Previous reports provided convincing information of higher PA and calcium absorption and efficient use of dietary energy when at this specific position in the TAG moiety than when PA is at the sn‐1,3 positions. During the design of structured lipids (SLs) intended for infant nutrition, this unique property is taken into consideration. Human milk fat substitutes (HMFS) enriched with important fatty acids such as omega‐3 and omega‐6 fatty acids are intended to better mimic the functions of HMF as well as provide associated health benefits. The use of microencapsulation technology and novel technologies such as ultrasound technology in conjunction with SL production and enzyme‐catalyzed reactions are evolving and ongoing issues in infant formula production. Therefore, further studies should be directed towards new process improvements in order to increase the functional properties and oxidative stabilities of HMFS. Novel technologies in lipid biotechnology related to HMFS preparation should also be explored.

Profiling of triacylglycerol composition in arachidonic acid single cell oil from Mortierella alpina by using ultra-performance liquid chromatography-electrospray ionization-quadrupole-time-of-flight mass spectrometry

Despite the increasing interest in arachidonic acid single-cell oil (ARA-SCO), the information about its triacylglycerol (TAG) composition still limited, keeping in mind that the profiling of TAG composition considered a challenging task due to the complexity of its structure. For tackling such a task, a comprehensive characterization of TAG molecular species in two different samples of ARA-SCO from Mortierella alpina was performed by using ultra-performance liquid chromatography with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). In the first sample, a total of 77 different TAGs were identified, while 73 TAGs were identified in the other sample. The most abundant TAG species in the first sample were (20:4-20:4-20:4), (20:3-20:4-16:0), (20:4-16:0-20:4), (18:1-16:0-20:3) and (24:0-20:3-18:1) with a relative content of 13.36, 8.43, 6.63, 4.52 and 4.27%, respectively. While (20:4-20:4-20:4), (20:4-24:0-20:4), (20:3-20:4-16:0), (20:4-16:0-20:4) and (18:1-16:0-20:3) were the major species in the second sample with a relative content of 11.47, 8.65, 7.84, 6.34 and 4.31%, respectively. Moreover, arachidonic acid, lignoceric acid, and palmitic acid were the predominant fatty acids in both ARA-SCO samples. This is the first work that reported an extensive characterization of TAG fractions of ARA-SCO, which might represent the key point for further studies and applications in the field of structured lipids and lipidomics.

Dietary Intake of Structured Lipids with Different Contents of Medium-Chain Fatty Acids on Obesity Prevention in C57BL/6J Mice.

Three medium- and long-chain triacylglycerols (MLCT) with different contents of medium-chain fatty acids (MCFA) (10% to 30%, w/w) were prepared and evaluated for their anti-obesity potential in C57BL/6J mice. The group fed with a high fat diet of MLCT containing 30% (w/w) MCFA showed significantly decreased body weight and fat mass (P < 0.05) relative to the control mice fed an obesity-inducing high fat rapeseed oil diet. In addition, serum parameters including triacylglycerols, total cholesterol, glucose, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, apolipoprotein A1 and apolipoprotein B in the treatment group fed with 30% (w/w) MCFA were close to those of mice fed with a low fat rapeseed oil diet, but significantly different (P < 0.05) from those of the obesity control group. Moreover, the intake of MLCT with high content of MCFA reduced the size of adipocytes. In addition, the visceral fat and liver weights, as well as the liver triacylglycerol for 3 treatment groups were lower than those of the obesity control group. These results demonstrate the great potential of MLCT with high content of MCFA in weight loss.

Synthesis of 1,3-dioleoyl-2-arachidonoylglycerol-rich structured lipids by lipase-catalyzed acidolysis of microbial oil from Mortierella alpina

Microbial oils (MOs) have gained widespread attention due to their functional lipids and health promoting properties. In this study, 1,3-dioleoyl-2-arachidonoylglycerol-rich structured lipids (SLs) were produced from MO and oleic acid (OA) via solvent-free acidolysis catalyzed by Lipozyme RM IM. Under the optimal conditions, the content of unsaturated fatty acids (UFAs) increased from 60.63 to 84.00%, while the saturated fatty acids (SFAs) content decreased from 39.37 to 16.00% at sn-1,3 positions in SLs. Compared with MO, arachidonic acid (ARA) content at the sn-2 position of SLs accounted for 49.71%, whereas OA was predominantly located at sn-1,3 positions (47.05%). Meanwhile, the most abundant triacylglycerol (TAG) species in SLs were (18:1-20:4-18:1), (20:4-20:4-18:1), (18:1-18:2-18:1), (18:1-18:2-18:0) and (24:0-20:4-18:1) with a relative content of 18.79%, 11.94%, 6.07%, 5.75% and 4.84%, respectively. Such novel SLs with improved functional properties enriched with UFAs are highly desirable and have the potential to be used in infant formula.

Production and characterization of structured lipids with antiobesity potential and as a source of essential fatty acids

The objective of this work was to produce structured lipids (SLs) from olive oil (O), soybean oil (S), and fully hydrogenated crambe oil – FHCO (C) mixtures by enzymatic interesterification, comparing Lipozyme TL IM and Rhizopus sp. performances as catalysts, and to evaluate their physical and chemical modifications. Among other blends (OC 90:10 w/w and SC 90:10 w/w), OSC (45:45:10, w/w), presented the most nutritionally interesting amounts of mono- and polyunsaturated fatty acids, as well as behenic acid. Interesterification caused an increase in crystallization time and a decrease in the solid fat content in all blends. The fatty acid redistribution in the TAGs caused a change in thermal behavior, leading to a decrease in the onset and end temperatures during crystallization, which indicates that new TAGs were formed. Regarding regiospecificity, Lipozyme TL IM lipase acted randomly, while Rhizopus sp. lipase was specific for the sn-1,3 position. Therefore, it was possible to synthesize SLs presenting different physical and chemical properties, compared to the original OSC blend, containing behenic acid at the sn-1,3 position and unsaturated fatty acids at the sn-2 position, by enzymatic interesterification catalyzed either by Lipozyme TL IM or by Rhizopus sp. lipases.

Production of structured lipid with a low omega-6/omega-3 fatty acids ratio by enzymatic interesterification

A structured lipid (SL) constituting omega fatty acids was synthesized by using linseed and grape seed oils as substrates via a lipase-catalyzed reaction. Lipozyme® TL IM was used as a biocatalyst. Good quadratic models predicting the incorporation of omega fatty acids were achieved via the Response surface methodology (RSM). The optimal conditions for targeted omega-6/omega-3 fatty acid ratio (2:1) were obtained at a substrate molar ratio 1.4, time 8.4 h, and enzyme amount 6.4%. The SL contained linoleic acid (43 g 100g-1), which was mainly located in the sn-2 position (40 g 100g-1). ?-Linoleic acid, and α-linolenic acid at the sn-2 position were 22 g 100g-1, and 11 g 100g-1, respectively. The oxidative stability of the SL, and SL with antioxidants was also investigated. The produced SL may be proposed as a source of a balanced intake of omega fatty acids and an ingredient in functional food formulations.

An efficient upgrading approach to produce n-3 polyunsaturated fatty acids-rich edible grade oil from high-acid squid visceral oil

High-acid squid visceral oil is unsuitable for consumption owing to its high content of free fatty acids (FAs). In this study, an edible grade resource of n-3 polyunsaturated fatty acids (PUFA) was developed from high-acid squid visceral oil (HASVO) by converting free FAs and partial glycerides in HASVO to ethyl esters (EEs) using a partial glycerides-selective lipase from Malassezia globosa while leaving triacylglycerol (TAG) intact. FAs in HASVO decreased from 13.84% to 0.06% corresponding to a deacidification efficiency of 99.57% under the optimal conditions. After purification by molecular distillation at 130°C, a final product contained 99.41% TAG, 0.51% diacylglycerol, and 0.08% FA was obtained with a TAG recovery of 98.86%. The acid, peroxide and anisidine value of the final product fulfilled the demand of edible grade and the high n-3 PUFA content (36.34%) in the obtained glyceride mixtures was comparable to other kinds of marine fish oil, suggesting it could be used as an alternative resource of n-3 PUFA. Moreover, the resulting byproduct EEs with 46.29% n-3 PUFA could be used as substrates for the synthesis of n-3 PUFA-rich structured lipids. This is the first study for upgrading of HASVO to edible grade with the form of TAG.

Enzymatic incorporation of caffeoyl into castor oil to prepare the novel castor oil-based caffeoyl structured lipids

In this work, a novel castor oil-based caffeoyl structured lipids was successfully prepared by the enzymatic transesterification using castor oil (CO) as caffeoyl acceptor. During the structured lipids preparation, two competitive reactions, the hydrolysis of CO to form hydrophilic caffeoyl glycerols (CG)+dicaffeoyl glycerols (DCG) and the transesterification of CO with ethyl caffeate (EC) to form lipophilic caffeoyl mono- and di-acylglycerols (CMAG and CDAG), were found. Reaction progress was monitored using HPLC-ESI–MS and HPLC-UV. The effects of by-product ethanol removal and reaction variables on the transesterification and reaction selectivity were evaluated. Results showed that, the activation energies for the transesterification and for the selective formations of CMAG+CDAG and CG+DCG were 57.60kJ/mol, 58.86kJ/mol, and 60.53kJ/mol, respectively. Under the optimal reaction conditions (enzyme load 23%, 90°C, 1:3 molar ratio of EC to CO, and 46.5h), EC conversion and the yield of CMAG+CDAG were 93.68±2.52% and 78.11±1.35%, respectively.

Evaluation of structured lipids with behenic acid in the prevention of obesity

Obesity affects all social classes, making it necessary to develop effective products that aid weight loss or help prevent weight gain. The objective of this work was to study the anti-obesity effects of structured lipids (SL) obtained by enzymatic interesterification, based on olive oil, soy oil and fully hydrogenated crambe oil. Twenty-four C57Bl/6 mice were distributed into four experimental groups according to the diet consumed: Control Diet (CD), Structured Lipids Diet (SLD), High-fat Control Diet (HCD), High-fat Structured Lipids Diet (HSLD). The animals that were fed SLs presented a smaller weight gain, despite a larger intake of the diet. The lowest weight gain was reflected in reduced amounts of adipose tissue and lower liver weight. A significant increase in lipids excreted by the animals in the feces was observed, despite there being no sign of toxicity or presence of diarrhea. The animals that consumed the HSLD presented lower total and LDL-cholesterol, increased HDL-cholesterol and increased hepatic arachidonic acid and docosahexaenoic acid levels. In addition, they did not develop hepatic steatosis. The study therefore showed that SLs could play a major role in combating or preventing obesity and other resultant diseases, without producing side effects.

Enzymatic Interesterification of Coconut and High Oleic Sunflower Oils for Edible Film Application

AbstractBlends [60:40, 70:30, and 80:20 (w/w)] of coconut oil (CO) and high oleic sunflower oil (HOSO) were interesterified using immobilized enzyme, Lipozyme® TL IM (Novozymes North America Inc., Franklinton, NC, USA). The structured lipids (SLs), referred to as interesterified products (IPs) IP60:40, IP70:30, and IP80:20, were compared to CO and HOSO for application in edible films. IPs were compared based on fatty acid profile, TAG molecular species, melting profile, moisture vapor permeability, mechanical properties, film transparency, density, and thickness. Interesterification increased oleic acid content at the sn‐2 position of IPs. CO had 5.50 ± 1.67 mol% oleic acid at the sn‐2 position, and when interesterified with HOSO (92.81 ± 1.10 mol% oleic acid) the amount of oleic acid significantly increased (p &lt; 0.05) at the sn‐2 position for IP60:40, IP70:30, and IP80:20 (33.86 ± 1.55, 27.34 ± 1.20, 20.61 ± 1.50 mol%), respectively. There was no significant difference between SLs, HOSO, and CO for water vapor permeability and density when applied to emulsion edible films. The HOSO film was significantly different (1.43 ± 0.27 AUmm−1) from the rest of the SLs and CO for film transparency. IP60:40 (2.20 ± 0.22 AUmm−1) decreased the opacity and was significantly different from HOSO and IP80:20 (2.88 ± 0.08 AUmm−1). Tensile strength of IP60:40 was 0.39 ± 0.17 MPa which was significantly different from IP70:30, IP80:20, and HOSO. The elongation at break was significantly different for HOSO and IP60:40. IP60:40 could be used to further investigate the use of SL in edible film for sports nutrition products.

Synthesis of structured lipids containing behenic acid from fully hydrogenated Crambe abyssinica oil by enzymatic interesterification.

Low-calorie structured lipids (SLs) rich in behenic and oleic acids were produced by enzymatic (EI) and chemical interesterification (CI) of high oleic sunflower oil and fully hydrogenated Crambe abyssinica oil in proportions of 60:40, 50:50, 40:60 and 30:70 (w/w), respectively. The immobilized lipase from Thermomyces lanuginosus (Lipozyme TL IM) was used for 3h at 70°C at a dosage of 7% (w/w) free of organic solvents. The original blend (BE) and the reaction products were evaluated for their fatty acid (FA) and triacylglycerols (TAG) compositions, solid fat contents, thermal analysis, regiospecific distribution of the FA, microstructure and polymorphism. The tendency was towards a reduction in the C52, C54, C60, C62 and C64 TAG contents and an increase in the C56, C58 e C66 TAG contents, therefore the solid fat content was reduced. There was lower acyl migration at the sn-2 position of the TAGs in EI as compared to CI in all the blends ratios. Needle-like crystals were predominant in the IE while large symmetrical spherulites were observed in the BE. Reduction in the mean crystal diameter was observed in all the blends, in addition to modifications of the crystal morphology. X-ray diffraction analysis showed a predominance of the β' form crystals in both the IE and CI. The SLs were produced for application in the food industry as bakery/confectionery fats in the proportions of 60:40/50:50 and as additive in the crystallization of lipids in the proportions of 40:60/30:70.

Development of an up-grading process to produce MLM structured lipids from sardine discards.

The aim of the work was to produce MLM structured lipids with caprylic acid (M) as medium chain fatty acid located at the external bonds of the glycerol backbone and concentrated polyunsaturated fatty acids (L) from sardine discards (Sardine pilchardus) in the central bond of the glycerol. To that end, the following steps were conducted: (i) fish oil extraction, (ii) Omega-3 free fatty acids (FFA) concentration (low temperature winterization), (iii) two-steps enzymatic esterification and (iv) triacylglycerols (TAG) purification (liquid column chromatography). The resultant purified triacylglycerols accomplished with the oxidative state (peroxide and anisidine value, PV and AV) required for refined oils. As enzymatic treatment, Omega-3 concentrate FFA (Omega-3>600mg Omega-3 per g oil) were esterified with dicaprylic glycerol employing Novozyme 435. This process presented high regioselectivity, with ∼80mol% of concentrated fatty acids esterified at the sn-2 position.

Correction: Analysis of moisture diffusion mechanism in structured lipids using magnetic resonance imaging

Correction for ‘Analysis of moisture diffusion mechanism in structured lipids using magnetic resonance imaging’ by Sravanti Paluri et al., RSC Adv., 2015, 5, 76904–76911.

Stability and Bioaccessibility of Fucoxanthin in Nanoemulsions Prepared from Pinolenic Acid-contained Structured Lipid

Abstract: Fucoxanthin intake has been correlated with the functions of anti-obesity and anti-oxidation, but applications of it in functional food or dietary supplements are still challenging due to its poor water-solubility, chemical instability, and low bioavailability. In this work, to study physicochemical and biological properties of fucoxanthin nanoemulsions, we investigated the influence of emulsion particle diameter on the stability of fucoxanthin during storage time and bioaccessibility in-vitro digestion. The structured lipid that enriched pinolenic acid at sn-2 position was chosen as the oil phase and the fucoxanthin oil-in-water nanoemulsions with droplet diameters of 344, 173, and 98 nm were prepared through a high-pressure microfluidizer. Then fucoxanthin emulsions were stored for 28 days at 4, 37, and 55 °C. Results showed that the physical stabilities of droplets were decreased with increases in the initial size and storage temperature, while the change of fucoxanthin retention indicated that fucoxanthin chemical stability was improved with increasing emulsion particle size. The augmentation of lipolysis and the value of free fatty acids (FFA) released in vitro digestion proved that digestion stability of fucoxanthin emulsion reduced with decreasing initial particle diameter, which was probably attributed to the increased surface area interacting with pancreatic lipase with decreasing droplet size. In addition, the concentrations of fucoxanthin in micelle phase were appreciable increased as droplet size decreased. Therefore, the bioaccessibility of fucoxanthin was improved. These results may benefit the optimization of an emulsion-based delivery system for fucoxanthin in food applications.

Quality by Design (QbD)-enabled development of aceclofenac loaded-nano structured lipid carriers (NLCs): An improved dermatokinetic profile for inflammatory disorder(s)

Present study was designed to prepare and characterize aceclofenac loaded nanostructured lipid carriers (NLCs) employing Quality by Design (QbD)-oriented approach. The NLCs were evaluated for their transdermal penetration potential and stability. Aceclofenac loaded nanostructured lipid carriers (NLCs) were prepared & characterized, by employing Quality by Design (QbD)-oriented approach and further evaluated for transdermal penetration potential and stability. Different lipids and surfactants were chosen to prepare NLCs using microemulsion method as critical material attributes (CMAs). A 33 factorial design was used for optimization of NLCs, and evaluating them for different critical quality attributes (CQAs), viz. particle size, polydispersity index (PDI), zeta potential, in vitro drug release, entrapment efficiency. The effect of CMAs such as lipids, oil: lipid ratio and concentration of surfactants on CQAs viz. drug entrapment efficiency and particle size were systematically evaluated to optimize NLCs. The optimized NLCs were further incorporated into carbopol gel and characterized for texture and rheology profile followed by in vitro and in vivo evaluations. The optimized ACE-NLCs were found to be spherical, nanometric in size with higher drug loading and entrapment efficiency. Results of the in vitro drug release study showed that the developed formulation followed Korsmeyer-Peppas model showing Fickian diffusion. The release was biphasic i.e., initial burst release followed by sustained drug release upto 48h. The optimized NLCs-based gel formulation showed superior texture, rheological profile and showed better cell uptake efficiency on hyperkeratinocytic cells (HaCaT cell lines) with higher ex vivo skin permeability efficiency vis-à-vis marketed formulation. In conclusion, dermatokinetic modeling and pharmacodynamic study using carrageenan induced edema mice suggests that aceclofenac loaded NLCs hydrogel may provide a better delivery alternative to target various skin layers.