Crystalline Cholesterol Research Articles

Lyotropic Liquid Crystalline Phase Nanostructure and Cholesterol Enhance Lipid Nanoparticle Mediated mRNA Transfection in Macrophages

AbstractMacrophages are unique immune cells attracting growing attention as a potential candidate for cell‐based therapy for infectious diseases and cancer. Strategies that can reprogramme or gene‐edit macrophages hold potential across a spectrum of acute and chronic conditions. Herein, lipid nanoparticles (LNPs) are developed containing the ionizable lipid SM‐102, helper lipid monoolein which is known for self‐assembly in aqueous solutions into the inverse cubic lyotropic liquid crystalline mesophase, and cholesterol as an mRNA nanocarrier. The immortalized alveolar macrophage cell line (MH‐S cells) is utilized to investigate how cholesterol concentration impacts on mRNA delivery which is further validated using primary mouse alveolar macrophages isolated from the bronchoalveolar compartment and human monocyte derived macrophages. By using high‐throughput synchrotron small angle X‐ray scattering (SAXS), an acidification‐induced non‐ordered to ordered internal nanostructure transition of the formulated LNPs is observed, following the transition sequence of inverse micellar to hexagonal to cubic mesophase in the pH range from 7 to 4. Cholesterol is identified as another crucial component for superior mRNA transfection in macrophages, contributing to nanostructure transition and protein corona variation. Successful ex vivo mRNA transfection is also achieved in primary macrophages, highlighting the prospectivity of reprogramming macrophages as a cell therapy for lung diseases.

Chemical potential measurements constrain models of cholesterol-phosphatidylcholine interactions

Bilayer membranes composed of cholesterol and phospholipids exhibit diverse forms of nonideal mixing. In particular, many previous studies document macroscopic liquid-liquid phase separation as well as nanometer-scale heterogeneity in membranes of phosphatidylcholine (PC) lipids and cholesterol. Here, we present experimental measurements of cholesterol chemical potential (μc) in binary membranes containing dioleoyl PC (DOPC), 1-palmitoyl-2-oleoyl PC (POPC), or dipalmitoyl PC (DPPC), and in ternary membranes of DOPC and DPPC, referenced to crystalline cholesterol. μc is the thermodynamic quantity that dictates the availability of cholesterol to bind other factors, and notably must be equal between coexisting phases of a phase separated mixture. It is simply related to concentration under conditions of ideal mixing, but is far from ideal for the majority of lipid mixtures investigated here. Measurements of μc can vary with phospholipid composition by 1.5 kBT at constant cholesterol mole fraction implying a more than fivefold change in its availability for binding receptors and other reactions. Experimental measurements are fit to thermodynamic models including cholesterol-DPPC complexes or pairwise interactions between lipid species to provide intuition about the magnitude of interactions. These findings reinforce that μc depends on membrane composition overall, suggesting avenues for cells to alter the availability of cholesterol without varying cholesterol concentration.

Freestanding Helical Nanostructured Chiro-Photonic Crystal Film and Anticounterfeiting Label Enabled by a Cholesterol-Grafted Light-Driven Molecular Motor.

Design and fabrication of freestanding chiro-photonic crystal film with the ability to change color over the whole visible light spectrum is appealing for anticounterfeiting technology and smart labels. Utilizing a newly synthesized light-responsive molecular motor functionalized with cholesterol (chol-MM) on the rotor, novel light-controlled photonic crystal is prepared by doping the novel chol-MM into liquid crystals (LCs). Thanks to the liquid crystalline cholesterol substituent, the chol-MM can be triggered by visible light (420nm). At the same time, the miscibility of chol-MM in LC matrix is significantly enhanced. Integrating the chol-MM with thermochromic hydrogen-bonded LC matrix, thermal and light dual-responsive cholesteric LC (CLC) material is prepared, in which the nanoscale helical pitch is tunable by photo-induced molecular motions of chol-MM. More importantly, utilizing UV-initiated polymerization of the visible light-modulated CLC material, structural colored photonic crystal films with arbitrary colorful patterns are fabricated. Such freestanding helical nanostructured labels have potential in the application of encrypted communication and anticounterfeiting.

Crystalline Cholesterol: The Material and Its Assembly Lines

Cholesterol is an essential component of animal cell membranes because it influences and controls cell membrane fluidity. Cholesterol is also responsible for the most frequent lethal pathologies in developed countries because of its intimate association with atherosclerotic plaques, the rupture of which may cause heart attacks or strokes. The question is under which conditions cholesterol activity manifests itself, whether in physiology or in pathology. The answer is complex, and there is probably not one certain answer. This review article has its foundations in abundant published knowledge and evidence, but it cannot possibly be comprehensive, because the extent of cholesterol's involvement in chemistry, biology, biophysics, and medicine is so vast that we cannot embrace it all. We review cholesterol as a molecule and in its various crystalline polymorphs. We then examine cholesterol assembly pathways and, finally, cholesterol in biology and in pathology. We propose that cholesterol activity depends on its assembly states in cholesterol crystals or with other lipids in the form of more-or-less organized crystalline domains. In other words, we analyze cholesterol material properties because the assembly state of the cholesterol molecules profoundly affects the properties of the environment in which they reside.

Cholesterol sulfate fluidizes stratum corneum lipid models and induces higher permeability

The stratum corneum (SC) is the outermost layer of the epidermis and protects the body from excessive water loss and xenobiotics. It is composed of corneocytes embedded in a continuous lipid matrix. This matrix is mandatory for the barrier function of the skin and consists of mainly crystalline free fatty acids, ceramides and cholesterol/-esters in a 1:1:1 molar ratio. Cholesterol sulfate (CSO4) as the most abundant ester of cholesterol (Chol) with up to 15 mol% of the total amount of sterols presents a lipid with a big polar headgroup in contrast to most of the other lipids in the SC.

The role of structural order in heterogeneous ice nucleation.

The freezing of water into ice is a key process that is still not fully understood. It generally requires an impurity of some description to initiate the heterogeneous nucleation of the ice crystals. The molecular structure, as well as the extent of structural order within the impurity in question, both play an essential role in determining its effectiveness. However, disentangling these two contributions is a challenge for both experiments and simulations. In this work, we have systematically investigated the ice-nucleating ability of the very same compound, cholesterol, from the crystalline (and thus ordered) form to disordered self-assembled monolayers. Leveraging a combination of experiments and simulations, we identify a “sweet spot” in terms of the surface coverage of the monolayers, whereby cholesterol maximises its ability to nucleate ice (which remains inferior to that of crystalline cholesterol) by enhancing the structural order of the interfacial water molecules. These findings have practical implications for the rational design of synthetic ice-nucleating agents.

The Initial Human Atherosclerotic Lesion and Lipoprotein Modification-A Deep Connection.

Atherosclerosis research typically focuses on the evolution of intermediate or advanced atherosclerotic lesions rather than on prelesional stages of atherogenesis. Yet these early events may provide decisive leads on the triggers of the pathologic process, before lesions become clinically overt. Thereby, it is mandatory to consider extracellular lipoprotein deposition at this stage as the prerequisite of foam cell formation leading to a remarkable accumulation of LDL (Low Density Lipoproteins). As progression of atherosclerosis displays the characteristic features of a chronic inflammatory process on the one hand and native LDL lacks inflammatory properties on the other hand, the lipoprotein must undergo biochemical modification to become atherogenic. During the last 25 years, evidence was accumulated in support of a different concept on atherogenesis proposing that modification of native LDL occurs through the action of ubiquitous hydrolytic enzymes (enzymatically modified LDL or eLDL) rather than oxidation and contending that the physiological events leading to macrophage uptake and reverse transport of eLDL first occur without inflammation (initiation with reversion). Preventing or reversing initial atherosclerotic lesions would avoid the later stages and therefore prevent clinical manifestations. This concept is in accordance with the response to retention hypothesis directly supporting the strategy of lowering plasma levels of atherogenic lipoproteins as the most successful therapy for atherosclerosis and its sequelae. Apart from but unquestionable closely related to this concept, there are several other hypotheses on atherosclerotic lesion initiation favoring an initiating role of the immune system (‘vascular-associated lymphoid tissue’ (VALT)), defining foam cell formation as a variant of lysosomal storage disease, relating to the concept of the inflammasome with crystalline cholesterol and/or mitochondrial DAMPs (damage-associated molecular patterns) being mandatory in driving arterial inflammation and, last but not least, pointing to miRNAs (micro RNAs) as pivotal players. However, direct anti-inflammatory therapies may prove successful as adjuvant components but will likely never be used in the absence of strategies to lower plasma levels of atherogenic lipoproteins, the key point of the perception that atherosclerosis is not simply an inevitable result of senescence. In particular, given the importance of chemical modifications for lipoprotein atherogenicity, regulation of the enzymes involved might be a tempting target for pharmacological research.

Implications of surfactant hydrophobic chain architecture on the Surfactant-Skin lipid model interaction

Although surfactants have been widely used in skin care and other related applications, our knowledge about how surfactants interact with stratum corneum (SC) lipids remains limited. This work reports how surfactants interact with a lipid SC model by neutron diffraction and molecular dynamics (MD) simulations, focusing on examining the impact of surfactant molecular architecture. The surfactant-SC mixed membrane was constructed by an equimolar mixture of ceramide/cholesterol/fatty acids and surfactant at 1% molar ratio of total lipids. The arrangements of water and surfactant molecules in the membrane were obtained through neutron scattering length density (NSLD) profiles via contrast variation method, meanwhile, MD simulation clearly demonstrated the mechanism of hydration change in the surfactant-model SC mixed membrane. No drastic difference was detected in the repeating distance of the short periodicity phase (SPP) upon adding surfactants, however, it significantly enhanced the membrane hydration and reduced the amount of phase separated crystalline cholesterol, showing a strong dependence on surfactant chain length, branching and double bond. This work clearly demonstrates how surfactant architecture affects its interaction with the SC membrane, providing useful guidance for either choosing an existing surfactant or designing a new one for surfactant-based transdermal application.

Non-linear optical imaging of atherosclerotic plaques in the context of SIV and HIV infection prominently detects crystalline cholesterol esters.

Chronic HIV infection may exacerbate atherosclerotic vascular disease, which at advanced stages presents as necrotic plaques rich in crystalline cholesterol. Such lesions can catastrophically rupture precipitating myocardial infarct and stroke, now important causes of mortality in those living with HIV. However, in this population little is known about plaque structure relative to crystalline content and its chemical composition. Here, we first interrogated plaque crystal structure and composition in atherosclerotic SIV-infected macaques using non-linear optical microscopy. By stimulated Raman scattering and second harmonic generation approaches both amorphous and crystalline plaque lipid was detected and the crystal spectral profile indicated a cholesterol ester (CE) dominated composition. Versus controls, SIV+ samples had a greater number of cholesterol crystals (CCs), with the difference, in part, accounted for by crystals of a smaller length. Given the ester finding, we profiled HIV+ plaques and also observed a CE crystalline spectral signature. We further profiled plaques from Ldlr-/- mice fed a high fat diet, and likewise, found CE-dominate crystals. Finally, macrophage exposure to CCs or AcLDL induced auto-fluorescent puncta that co-stained with the LC3B autophagy sensor. In aggregate, we show that atheromatous plaques from mice, macaques and humans, display necrotic cores dominated by esterified CCs, and that plaque macrophages may induce autophagic vesicle formation upon encountering CCs. These findings help inform our knowledge of plaque core lipid evolution and how the process may incite systemic inflammation.

Flexible pulse shaping for sum frequency microspectroscopies

The complexity of experimental schemes still challenges the broad application of multimodal microscopy beyond fundamental research. Pulse shaping has been a promising candidate to assist in the implementation of flexible and compact schemes. In this work, its application is demonstrated for vibrational resonant sum frequency (SF) spectroscopy in a nonlinear Raman microscope. The mixing of an ultrabroadband NIR laser pulse with a difference-frequency-generated MIR spectrum provides spectral access from 1750 c m − 1 to beyond 3000 c m − 1 via two shaping methods: (i) homodyne MIR scanning and (ii) heterodyne multiplex. The versatility of shaper-based SF is demonstrated in the spectroscopy and microscopy of crystalline cholesterol and cysteine.

PERFIL LIPÍDICO E GLICÊMICO DE RATOS SUBMETIDOS A DIETAS COM FIBRA DA CASCA E TALO DE BANANA

Objetivo: O estudo teve por objetivo avaliar os efeitos da ingestão de dietas contendo a fibra presente na casca e talo da banana, no perfil glicêmico e lipídico de ratos da linhagem Wistar. Metodologia: Estudou-se, durante 65 dias, a influência de dietas contendo fibras provenientes da casca (GFC), talo da banana (GFT) e ambas as fibras (GFCT), em relação à dieta controle (GA), no perfil glicêmico e lipídico de 24 ratos machos da linhagem Wistar, com peso médio inicial de 54,06g, distribuídos de forma equitativa em quatro grupos. O grupo controle (GA) recebeu dieta AIN acrescida de 1% de colesterol cristalino e 0,1% de ácido cólico, e as dietas experimentais foram semelhantes ao grupo controle, com substituição da fonte de fibra. Resultados: Nos grupos de tratamento (GFC, GFT e GFCT), houve aumento dos níveis glicêmicos, de triglicerídeos e redução do colesterol total e LDL-C séricos, em relação ao grupo controle (GA). Os níveis de VLDL-C mostraram-se reduzidos apenas para o grupo contendo fibras provenientes da casca. Conclusões: A inserção de fibra na dieta, proveniente da casca e talo da banana, bem como da mistura de ambos, mostrou efetividade de uso sobre o perfil lipídico e capacidade de manutenção dos níveis de glicemia dos ratos alimentados com dietas contendo fibra oriunda da casca e do talo.DOI: 10.12957/demetra.2018.29559 DOI: 10.12957/demetra.2018.29559

Placental inflammation in pre-eclampsia by Nod-like receptor protein (NLRP)3 inflammasome activation in trophoblasts.

Pre-eclampsia is associated with increased levels of cholesterol and uric acid and an inflamed placenta expressing danger-sensing pattern recognition receptors (PRRs). Crystalline cholesterol and uric acid activate the PRR Nod-like receptor protein (NLRP)3 inflammasome to release interleukin (IL)-1β and result in vigorous inflammation. We aimed to characterize crystal-induced NLRP3 activation in placental inflammation and examine its role in pre-eclampsia. We confirmed that serum total cholesterol and uric acid were elevated in pre-eclamptic compared to healthy pregnancies and correlated positively to high sensitivity C-reactive protein (hsCRP) and the pre-eclampsia marker soluble fms-like tyrosine kinase-1 (sFlt-1). The NLRP3 inflammasome pathway components (NLRP3, caspase-1, IL-1β) and priming factors [complement component 5a (C5a) and terminal complement complex (TCC)] were co-expressed by the syncytiotrophoblast layer which covers the placental surface and interacts with maternal blood. The expression of IL-1β and TCC was increased significantly and C5a-positive regions in the syncytiotrophoblast layer appeared more frequent in pre-eclamptic compared to normal pregnancies. In-vitro activation of placental explants and trophoblasts confirmed NLRP3 inflammasome pathway functionality by complement-primed crystal-induced release of IL-1β. This study confirms crystal-induced NLRP3 inflammasome activation located at the syncytiotrophoblast layer as a mechanism of placental inflammation and suggests contribution of enhanced NLRP3 activation to the harmful placental inflammation in pre-eclampsia.

OP 28 Inflammation in preeclampsia by nod-like receptor protein (nlrp)3 inflammasome activation in trophoblasts

Introduction Preeclampsia (PE) is characterized by an aberrant inflammatory response to pregnancy. A stressed and inflamed placenta will induce maternal systemic inflammation, leading to hypertension with proteinuria (i.e. PE). The harmful placental inflammation involves abnormal placental expression of danger-sensing pattern recognition receptors (PRRs). Increased maternal cholesterol and uric acid levels are also associated with PE development. Crystalline cholesterol and uric acid may activate the PRR named nod-like receptor protein (NLRP)3 inflammasome to release the potent inflammatory cytokine IL-1 β , resulting in vigorous inflammation. Objective We aimed to characterize crystal-induced NLRP3 activation in placental inflammation, and examine its role in PE development. Materials and methods Total cholesterol, uric acid, soluble fms-like tyrosine kinase-1 (sFlt-1) and high sensitivity C-reactive protein (hsCRP) were measured in serum from pregnant women with (n = 34) or without (n = 43) PE and non-pregnant women (n = 28). Specific cell markers, central NLRP3 inflammasome pathway components (NLRP3, caspase 1 and IL-1 β ) , and complement factors (C5a and TCC) were detected by immunohistochemistry in third trimester placental samples from healthy (n = 13) and PE pregnancies (n = 23). Placental explants and the trophoblast cell line SGHPL-5 were primed with lipopolysaccharide (LPS) or C5a/tumor necrosis factor (TNF)- α and stimulated with cholesterol or uric acid crystals. Resulting IL-1 β response was quantified by ELISA. Results Cholesterol and uric acid levels were elevated in maternal serum from PE compared to healthy pregnancies and correlated with levels of hsCRP and sFlt-1. The NLRP3 inflammasome components and complement factors were expressed by placental syncytiotrophoblast (SCT), and the IL-1 β and TCC expression in SCT was significantly higher in PE compared to healthy pregnancies. C5a positive regions of the SCT were observed more frequently in PE placentas. Complement primed crystal stimulation induced NLRP3 dependent IL-1 β production in trophoblasts and explants cultures.

NLRP3 inflammasome: Its regulation and involvement in atherosclerosis.

Inflammasomes are intracellular complexes involved in the innate immunity that convert proIL-1β and proIL-18 to mature forms and initiate pyroptosis via cleaving procaspase-1. The most well-known inflammasome is NLRP3. Several studies have indicated a decisive and important role of NLRP3 inflammasome, IL-1β, IL-18, and pyroptosis in atherosclerosis. Modern hypotheses introduce atherosclerosis as an inflammatory/lipid-based disease and NLRP3 inflammasome has been considered as a link between lipid metabolism and inflammation because crystalline cholesterol and oxidized low-density lipoprotein (oxLDL) (two abundant components in atherosclerotic plaques) activate NLRP3 inflammasome. In addition, oxidative stress, mitochondrial dysfunction, endoplasmic reticulum (ER) stress, and lysosome rupture, which are implicated in inflammasome activation, have been discussed as important events in atherosclerosis. In spite of these clues, some studies have reported that NLRP3 inflammasome has no significant effect in atherogenesis. Our review reveals that some molecules such as JNK-1 and ASK-1 (upstream regulators of inflammasome activation) can reduce atherosclerosis through inducing apoptosis in macrophages. Notably, NLRP3 inflammasome can also cause apoptosis in macrophages, suggesting that NLRP3 inflammasome may mediate JNK-induced apoptosis, and the apoptotic function of NLRP3 inflammasome may be a reason for the conflicting results reported. The present review shows that the role of NLRP3 in atherogenesis can be significant. Here, the molecular pathways of NLRP3 inflammasome activation and the implications of this activation in atherosclerosis are explained.

Recombinant anthrax protective antigen: Observation of aggregation phenomena by TEM reveals specific effects of sterols

Negatively stained transmission electron microscope images are presented that depict the aggregation of recombinant anthrax protective antigen (rPA83 monomer and the PA63 prepore oligomer) under varying in vitro biochemical conditions. Heat treatment (50°C) of rPA83 produced clumped fibrils, but following heating the PA63 prepore formed disordered aggregates. Freeze-thaw treatment of the PA63 prepore generated linear flexuous aggregates of the heptameric oligomers. Aqueous suspensions of cholesterol microcrystals were shown to bind small rPA83 aggregates at the edges of the planar bilayers. With PA63 a more discrete binding of the prepores to the crystalline cholesterol bilayer edges occurs. Sodium deoxycholate (NaDOC) treatment of rPA83 produced quasi helical fibrillar aggregate, similar but not identical to that produced by heat treatment. Remarkably, NaDOC treatment of the PA63 prepores induced transformation into pores, with a characteristic extended ß-barrel. The PA63 pores aggregated as dimers, that aggregated further as angular chains and closed structures in higher NaDOC concentrations. The significance of the sterol interaction is discussed in relation to its likely importance for PA action in vivo.

Inflamation Leads to an Increase of Cholesterol's Chemical Potential in Plasma Membranes of Cultured Cells

Our laboratory has developed a method to measure the chemical potential of cholesterol in cell membranes. We have begun to use the procedure to explore the consequences of cell inflammation on chemical potential. The chemical potential of membrane cholesterol in non-inflamed primary dermal human fibroblasts is ≈ −2.3 kBT relative to crystalline cholesterol. Treating these cells with tumor necrosis factor (TNF-α) for 72 hr results in an increase of the chemical potential. This increase occurs in a dose-dependent manner: modest for 10 ng/ml TNF-α, about 0.8 kBT greater for 20 ng/ml, and only slightly greater for 40 ng/ml. Simultaneous treatment with TNF-α and 50 nM of the anti-inflammatory agent dexamethasone abolishes the increase in the chemical potential of cholesterol caused by TNF-α alone. Using 5 ng/ml of interleukin-1β, instead of TNF-α, as a pro-inflammatory agent also resulted in a rise of chemical potential. But the effect was less than that induced by TNF-α; the increase was ≈ 0.5 kBT. These results indicate a possible causative relation between cell inflammation and the chemical potential of plasma membrane cholesterol. Supported by R01 GM101539.

Antifriction Plastic Greases Based on Intermediate Products of Oil Refining and Liquid Crystalline Compounds

The rheological and triboengineering properties are presented of hydrated calcium plastic greases based on intermediate products of oil refining (hydrofined vacuum gas oil and vacuum distillate). It is shown that application of liquid crystalline cholesterol compounds as an additional component of the dispersion medium of plastic greases permits significant reduction of the wear and losses for friction and boosts the load – bearing capacity of tribocouples.

Abstract 447: β-cyclodextrin Reduces Cholesterol Crystal-induced Inflammation Through Modulating Complement Activation

Atherosclerosis is an inflammatory condition and the underlying cause for cardiovascular disease. Cholesterol crystals (CC) are found to be abundant in atherosclerotic plaques and we have previously shown that CC initiate an inflammatory response via the complement system and inflammasome activation. Cyclic oligosaccharide 2-hydroxypropyl-β-cyclodextrin (BCD) is a compound that solubilizes lipophilic substances and is commonly used in pharmaceuticals or drug delivery. BCD is reported to increase cholesterol solubility and to promote the removal of cholesterol from foam cells. However, it remains unknown whether BCD has any effect on crystalline cholesterol. We here show that BCD attenuates the CC -induced inflammatory cytokine response as well as regulates a range of CC-related genes in human peripheral blood mononuclear cells. BCD binds to CC and prevents deposition of complement factors on CC in human plasma. Furthermore, BCD also decreases the formation of soluble terminal complement complex (TCC) and the expression of complement receptor 3 in response to CC stimulation in human whole blood. Induction of TCC by mono sodium urate crystals or zymosan was not affected by BCD. Of interest, after 1 hr of incubation, BCD is starting to dissolve the CC. These data demonstrate that BCD is a strong inhibitor of CC-induced inflammation, which might be explained by BCD-mediated attenuation of complement activation. These data suggest that BCD is a potential candidate for treatment of atherosclerosis.

The Chemical Potential of Cholesterol Regulates the Pro-Metastatic Phenotype in a Cell Culture Model of Breast Cancer

We have developed a simple and reliable method to determine the chemical potential of cholesterol in plasma membranes of living cells in vitro. For a variety of cultured cells, including non-metastatic breast cancer cells, the chemical potential of cholesterol is maintained at a level of about −1.9 kBT per moleculerelative to crystalline cholesterol. But for a metastatic breast cancer cell line, this chemical potential is appreciably greater, at about −0.7 kBT per molecule. In light of these observations and recent reports of cholesterol's role in breast cancer progression we have developed a method to alter and maintain the chemical potential of cholesterol in plasma membranes of cultured cells at any desired level. Through this technique, we lowered the chemical potential of cholesterol in the metastatic cell line to that of the non-metastatic line and found that this significantly reduced the expression levels of several key proteins implicated in breast cancer progression and metastatic spread.

The Development of Diets to Induce Atherogenic Lipid Profiles for Cynomolgus Monkeys in Their Country of Origin

Understanding the process of atherosclerosis progress can be studied in laboratory animals, such as nonhuman primate (NHP)<i>.</i> Investigators at Bogor Agricultural University Indonesia, Primate Research Center (IPB) reported to develop an atherogenic diet (IPB 1) by using fresh egg yolk and coconut oil as source of cholesterol and fat. The aims of the research were to correct nutritional inadequacy in the initial IPB 1 atherogenic diet by supplementation with corn oil (IPB 1+CO); to use dry powdered egg yolk (PEY) instead of fresh egg yolk (IPB 1+CO+PEY); to use concentrated source of protein (43%) pupae meal (PM) instead of soya meal (IPB 1+CO+PM) ; and to use crystalline cholesterol (CC) instead of egg yolk (IPB 1+CO+CC). Twenty four <i>Macaca fascicularis</i> were used as animal model for three months adaptation followed by 12 months for four diet treatments. Parameters measured such body weight, waist circumference, trunk length, adiposity index, nutrient utilization, and plasma lipid profile every three months. This experiment used Completely Randomized Design with four treatments and six replications. Result showed that there were no significant differences found in morphometric parameters among the diet groups compared to one another or change from baseline. The nutrient (protein, fat and carbohydrate) consumption and the absorption were essentially the same for all four diet groups. The IPB 1+CO diet, the IPB 1+CO+PM diet and the IPB 1+CO+CC induced a similar atherogenic plasma lipid profile, with marked increases in total plasma cholesterol concentrations.