Yolk Phosphatidylcholine Research Articles

Solubilization of egg phosphatidylcholine liposomes by sodium taurocholate: partition behavior and morphology

Solubilization process of egg yolk phosphatidylcholine (EggPC) liposomes by sodium taurocholate (TC) was performed using membrane dialysis technique. Turbidity, apparent particle size, Cl− permeability, membrane fluidity measured by electron spin resonance spectroscopy (ESR), and structural changes observed by freeze-fracture electron microscopy were investigated. The concentrations of TC were analyzed by high-performance liquid chromatography to determine the incorporation of TC in membrane phase and apparent partition coefficient. When TC concentration in water phase (Dw) was lower than 1.41 mM, or effective ratio of TC to EggPC (Re) was less then 0.14, the partition coefficient was independent of the TC concentration and was 90 M−1, where the membrane permeability of Cl− was rather small. Upon increasing the concentrations in the range of 1.41 mM≤Dw<3.10 mM (0.14≤Re<0.21), the apparent partition coefficient decreased and membrane permeability markedly increased. The coexistence of small vesicles and bilayer fragments were observed in this region. At 3.10 mM≤Dw<3.53 mM (0.21≤Re<0.59), the steep decrease in turbidity was accompanied with the gradual increase in apparent partition coefficient, suggesting the formation of mixed micelles. The freeze-fracture electron micrographs confirmed the transformation of saturated bilayers to wormlike mixed micelles. When a further increase in TC (Dw≥3.53 or Re≥0.59) was applied, the turbidity and apparent particle sizes became small. Transformation of the wormlike structure to ellipsoid mixed micelles was observed. The ESR spectra showed that TC/EggPC mixed micelles still retain a somewhat orderly structure.

Use of Phospholipid Transfer Protein as a Probe to Study the Lipid Dynamics and Alkaline Phosphatase Activity in the Brush Border Membrane of Human Term Placenta

Incubation of placental brush border membrane (BBM) along with sonicated vesicles of exogenous lipids (egg yolk PC) in the presence of phospholipid-transfer protein (PL-TP) showed a decrease in the alkaline phosphatase activity due to the change in the membrane micro-environment, such as fluidity. Effect of substrate concentration was tested by Lineweaver-Burk plot, which showed decreased V(max) and K(M). The effect of temperature was probed by the Arrhenius plot, which showed no change in transition temperature, but a decline in the energy of activation both below and above the transition temperature. The protein-catalyzed transfer of phospholipid from the donor unilamellar vesicles resulted in a substantial increase in the BBM phospholipid and a net decrease in cholesterol/phospholipid molar ratio. The change in membrane fluidity was assessed by translational as well as rotational diffusion of membrane extrinsic fluorescent probes, pyrene and diphenyl-hexatriene. An increased lateral mobility was recorded by the increased pyrene excimer formation. A decrease in fluorescent polarization of diphenyl-hexatriene was observed, which led to the decrease in fluorescence anisotropy and order parameter, and therefore, an increase in membrane fluidity (rotational diffusion). Mean anisotropy parameter was also decreased in the presence of PL-TP. Thus, the placental BBM alkaline phosphatase activity showed a distinct lipid dependence which may have important physiological consequences.

Polymeric Vesicle Permeability: A Facile Chemical Assay

We present a simple method to characterize vesicles and determine, at the same time, the membrane permeability to specific molecules. The method is based on encapsulating highly hydrophilic 3,3',3' '-phosphinidynetris-benzenesulfonic acid (PH) into vesicles and subsequently monitoring its reaction with 5,5'-dithiobis-2-nitrobenzoic acid (DTNB). We tested the method by measuring the membrane permeability of vesicles formed from a series of poly(ethylene oxide)-co-polybutylene oxide (EB) copolymers and egg yolk phosphatidylcholine. We found that the experimental data are in good agreement with calculations based on Fick's first law. We therefore quantified the DTNB permeability across vesicle membranes, finding that polymeric EB membranes have a more selective permeability toward polar molecules compared to phospholipids membranes.

Prolonged blood concentration of prednisolone after intravenous injection of liposomal palmitoyl prednisolone

We compared the pharmacokinetic behavior of drugs after an intravenous administration of prednisolone (PLS), palmitoyl prednisolone (Pal-PLS), and liposomal Pal-PLS in rats. Pal-PLS showed higher lipophilicity and higher binding to plasma protein than PLS, and PLS regeneration in rat blood and liver homogenates. After the intravenous administration of Pal-PLS solution in polyethylene glycol (PEG) 400 to rats, Pal-PLS disappeared from the blood in a two-phase mode and PLS was rapidly regenerated. Pal-PLS showed a significantly higher accumulation than PLS in the liver and lung. The administration of Pal-PLS incorporated into egg yolk phosphatidylcholine (EggPC)/cholesterol (Chol) liposomes enhanced Pal-PLS concentrations in the blood, liver, and lung compared to that of Pal-PLS solution in PEG 400, suggesting the rapid removal of liposomes by the mononuclear phagocytic system. Pal-PLS incorporated into PEGylated liposomes constituted with EggPC/Chol/1% L-α-distearoylphosphatidylethanolamine (DSPE)-PEG 2000 and EggPC/Chol/10% DSPE-PEG 2000 decreased the initial distribution of Pal-PLS, and successfully maintained the blood concentrations of Pal-PLS and PLS. Thus, we could change the pharmacokinetics of PLS by introducing the palmitoyl function into the molecule and its liposomal formulation including PEGylation. This is the first study to evaluate liposomal PLS constituted with a lipophilic derivative and PEG lipids.

Binding of antibiotic amphotericin B to lipid membranes: A 1H NMR study

The 1H NMR technique was applied to study binding of AmB, an antifungal drug, to lipid membranes formed with egg yolk phosphatidylcholine. The analysis of 1H NMR spectra of liposomes, containing also cholesterol and ergosterol (at 40 mol%), shows that AmB binds preferentially to the polar headgroups. Such a binding restricts molecular motion of the choline fragment in the hydrophilic region at the surface of liposomes but increases the segmental motional freedom in the hydrophobic core. The same effects are also observed in the sterol-containing membranes, except that the effect on the hydrophobic core was exclusively observed in the membranes containing ergosterol.

Stability of phospholipid vesicles studied by asymmetrical flow field-flow fractionation and capillary electrophoresis

The stability of zwitterionic phosphatidylcholine vesicles in the presence of 20 mol% phosphatidyl serine (PS), phosphatidic acid (PA), phosphatidyl inositol (PI), and diacylphosphatidyl glycerol (PG) phospholipid vesicles, and cholesterol or calcium chloride was investigated by asymmetrical flow field-flow fractionation (AsFlFFF). Large unilamellar vesicles (LUV, diameter 100 nm) prepared by extrusion at 25 °C were used. Phospholipid vesicles (liposomes) were stored at +4 and −18 °C over an extended period of time. Extruded egg yolk phosphatidylcholine (EPC) particle diameters at peak maximum and mean measured by AsFlFFF were 101 ± 3 nm and 122 ± 5 nm, respectively. No significant change in diameter was observed after storage at +4 °C for about 5 months. When the storage period was extended to about 8 months (250 days) larger destabilized aggregates were formed (172 and 215 nm at peak maximum and mean diameters, respectively). When EPC was stored at −18 °C, large particles with diameters of 700–800 nm were formed as a result of dehydration, aggregation, and fusion processes. In the presence of calcium chloride, EPC alone did not form large aggregates. Addition of 20 mol% of negatively charged phospholipids (PS, PA, PI, or PG) to 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphatidylcholine (POPC) vesicles increased the electrostatic interactions between calcium ion and the vesicles and large aggregates were formed. In the presence of cholesterol, large aggregates of about 250–350 nm appeared during storage at +4 and −18 °C for more than 1 day. The effect of liposome storage temperature on phospholipid coatings applied in capillary electrophoresis (CE) was studied by measuring the electroosmotic flow (EOF). EPC coatings with and without cholesterol, PS, or calcium chloride, prepared from liposomes stored at +25, +4, and −18 °C, were studied at 25 °C. The performances of the coatings were further evaluated with three uncharged compounds. Only minor differences were observed between the same phospholipid coatings, showing that phospholipid coatings in CE are relatively insensitive to storage at +25, +4 °C or −18 °C.

Germination as a process to improve the antioxidant capacity of Lupinus angustifolius L. var. Zapaton

Antioxidant capacity, measured by glutathione (GSH), superoxide dismutase-like activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), trolox equivalent antioxidant capacity (TEAC) and inhibition of lipid peroxidation in unilamellar liposomes of egg yolk phosphatidylcholine (PC) has been evaluated in raw and germinated lupin seeds (Lupinus angustifolius L. var. Zapaton) for 2, 3, 4, 5, 6 and 9 days. The content of antioxidant vitamins E and C has been also studied. The tripeptide GSH kept invariable for the first 5 days of germination and suffered a decrease of 20 and 78% after 6 and 9 days, respectively. During lupin germination, SOD-like activity increased slightly whilst PRTC doubled the amount after 9 days. TEAC values changed slightly up to 5 days of germination but after 6 and 9 days a significant increase (25 and 28%, respectively) was found. The oxidation of PC was inhibited by germinated lupin extracts and 9-day germination seeds provided the highest inhibition. Furthermore, germinated lupins provided more vitamin C, vitamin E activity and polyphenols than raw seeds, and the largest amounts of these bioactive compounds were found after 6 days of germination. Therefore, germination of lupin seeds (Lupinus angustifolius L. var. Zapaton) seems to be a good process to enhance their antioxidant capacity.

Enhanced Delivery of Retinoic Acid to Skin by Cationic Liposomes

We studied the delivery of retinoic acid to skin by using cationic liposomes consisting of double-chained cationic surfactant, phosphatidylcholine (PC) and retinoic acid in excised guinea pig dorsal skin. Egg yolk PC liposomes contaning retinoic acid at a molar ratio of 4 : 1 increased the delivery of retinoic acid about two-fold, compared with its addition as an isopropyl myristate solution. Cationic liposomes containing 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) further enhanced the incorporation dependent on the DOTAP content. Liposomes consisting of DOTAP, egg yolk PC, and retinoic acid at a molar ratio of 2 : 2 : 1 induced a 3.7-fold increase in the skin incorporation compared with the egg yolk PC liposomes without DOTAP. Significant difference was not observed when either dimyristoylphosphatidylcholine (DMPC) or dipalmitoylphosphatidylcholine (DPPC) was used instead of egg yolk PC as well as when dimethyldipalmitylammonium was used instead of DOTAP. These results suggest the potential of the use of the cationic liposomes for the intradermal delivery of lipophilic drugs like retinoic acid.

Characterization of phospholipid + semifluorinated alkane vesicle system

The aim of this study is to characterize vesicles obtained by the incorporation of the semifluorinated alkane, (perfluoro- n-hexyl)ethane (diblock F 6H 2) to a standard lipid, egg yolk phosphatidylcholine (PC). Large unilamellar vesicles (LUVs), prepared by extrusion, were characterized by fluorescence spectroscopy, zeta potential (ζ-potential) and light scattering. By using the fluorescence spectroscopy technique, the anisotropy of l,6-diphenyl-l,3,5-hexatriene (DPH) probe at different temperatures was determined. It was demonstrated that F 6H 2 is placed inside of the lipid bilayer and that the hydrocarbon acyl chain in the bilayers has higher viscosity in the presence of fluoroalkane. The ζ-potential of the PC–F 6H 2 system is negative and increases (in absolute value) from −10 to −19 mV when the temperature rises from 10 to 25 °C, this last value keeping practically constant with a further increase of temperature. The adsorption of K + ions on the liposome surface was measured by ζ-potential. This adsorption originates a sudden increase of the initial ζ-potential followed by a slight decrease with K + concentration. The application of the DLVO theory of colloidal stability showed a growing dependence of the DLVO potential with K + concentration and consequently a increasing stability.

Absorption and lipoprotein transport of sphingomyelin

Dietary sphingomyelin (SM) is hydrolyzed by intestinal alkaline sphingomyelinase and neutral ceramidase to sphingosine, which is absorbed and converted to palmitic acid and acylated into chylomicron triglycerides (TGs). SM digestion is slow and is affected by luminal factors such as bile salt, cholesterol, and other lipids. In the gut, SM and its metabolites may influence TG hydrolysis, cholesterol absorption, lipoprotein formation, and mucosal growth. SM accounts for approximately 20% of the phospholipids in human plasma lipoproteins, of which two-thirds are in LDL and VLDL. It is secreted in chylomicrons and VLDL and transferred into HDL via the ABCA1 transporter. Plasma SM increases after periods of large lipid loads, during suckling, and in type II hypercholesterolemia, cholesterol-fed animals, and apolipoprotein E-deficient mice. SM is thus an important amphiphilic component when plasma lipoprotein pools expand in response to large lipid loads or metabolic abnormalities. It inhibits lipoprotein lipase and LCAT as well as the interaction of lipoproteins with receptors and counteracts LDL oxidation. The turnover of plasma SM is greater than can be accounted for by the turnover of LDL and HDL particles. Some SM must be degraded via receptor-mediated catabolism of chylomicron and VLDL remnants and by scavenger receptor class B type I receptor-mediated transfer into cells.

Interaction of (+)-Catechin with a Lipid Bilayer Studied by the Spin Probe Method

The interaction of (+)-catechin with a lipid bilayer was examined by the spin probe method. The spin probe, 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO), was dissolved in an aqueous dipalmitoylphosphatidylcholine (DPPC) dispersion containing (+)-catechin. The temperature dependence of the TEMPO parameter was measured. The increase of this parameter due to pretransition was eliminated by the addition of (+)-catechin, suggesting that it was adsorbed to the lipid membrane surface in the gel state, which hindered the change of the membrane from a flat to wavy structure. In the temperature region of the main transition, the TEMPO parameter increased rapidly, then gradually with increasing temperature, which could be explained by the eutectic phase diagram. The rotational correlation time of a spin probe 16-doxylstearic acid and the order parameter of 5-doxylstearic acid in the aqueous dispersion system of egg yolk phosphatidylcholine revealed that the motion of the alkyl chain in the liquid crystal state was hindered in the center of the membrane as well as near the surface by the adsorption of (+)-catechin.

Effect of phosphatidylcholine vesicle size on chirality induction and chiral discrimination

The effect of the size of phosphatidylcholine (PC) vesicles on the induction of chirality and chiral discrimination was examined. Three kinds of vesicles formed with l-dimyristoyl, l-dipalmitoyl, or egg yolk PCs induced circular dichroisms (CDs) with the sign and intensity of the Cotton effect different from those of monomeric PCs. The CD intensity of the vesicles increased with a decrease in the vesicle size. Furthermore, the helicity of heterohelicene derivatives in a rapid equilibrium between right-handed (P) and left-handed (M) enantiomers was biased toward the M enantiomer side in l-PC vesicles, implying chiral discrimination by the vesicles. The extent of the bias toward the M enantiomer increased with an increase in vesicle size. Both the chirality induction and chiral discrimination were enhanced in a low-fluidity gel phase in comparison with those in a high-fluidity liquid-crystalline phase for every kind of vesicle of every size examined.

Inhibiting effect of tea catechins on the lipid peroxidation induced in tritiated water

Lipid peroxidation induced by β-ray in tritiated water and the inhibiting effect of tea catechins on it were studied using a spin probe method. A hydrophobic spin probe, 16-doxylstearic acid (16NS), was incorporated into a liposome prepared from egg yolk phosphatidylcholine, which was dispersed in tritiated water; the catechins were added to the solution. The rate of the decrease of ESR intensity of 16NS was a measure of the peroxidation and of the inhibiting effect. Inhibiting activity increased with an increase in the concentration of the catechin. Inhibiting ability estimated from the slope of the curves was in the order of (−)-epicatechin gallate > (−)-epigallocatechin gallate > (−)-epicatechin > (−)-epigallocatechin. The activity decreased with increasing temperature and the temperature dependence increased with the catechin concentration. These results were explained by a model; the initiator of the peroxidation is the hydroxyl radical (OH) and catechin is adsorbed on the surface of the membrane and scavenges OH coming into there from the water phase. The activity depended on the ratio of the adsorbed catechin, namely the partition coefficient between the water and the lipid.

Temperature Sensitization of Liposomes by Use of Thermosensitive Block Copolymers Synthesized by Living Cationic Polymerization: Effect of Copolymer Chain Length

We prepared block copolymers of (2-ethoxy)ethoxyethyl vinyl ether (EOEOVE) and octadecyl vinyl ether (ODVE) with the number average molecular weights of 6900, 9300, and 16 700 by living cationic polymerization. The poly(EOEOVE) block acts as a temperature-sensitive moiety, and the poly(ODVE) block acts as an anchor moiety. We also investigated the effect of chain length of the copolymer poly(EOEOVE) block on the ability to sensitize liposomes. The copolymers underwent a coil-globule transition at approximately 36 degrees C in the presence of a membrane of egg yolk phosphatidylcholine (EYPC), detected using differential scanning calorimetry (DSC). Liposomes encapsulating calcein, a water-soluble fluorescent dye, were prepared from mixtures of dioleoylphosphatidylethanolamine, EYPC, and the copolymers. While the copolymer-modified liposomes released little calcein below 30 degrees C, release was enhanced above 35 degrees C, indicating that dehydrated copolymer chains destabilized the liposome membrane. In addition, copolymers with a longer poly(EOEOVE) block induced a more drastic enhancement of contents release in a narrow temperature region near the transition temperature of the poly(EOEOVE) block. As a result, the copolymer with an average molecular weight of 16 700 generated highly sensitive liposomes that produced rapid and dramatic release of the contents in response to temperature.

Membrane Fluidity of Egg Yolk Phosphatidylcholine/Detergent Mixed Aggregates Studied by Spin Label Method

Abstract Anisotropic electron spin resonance (ESR) spectra of 5-doxylstearic acid and small membrane fluidity of hydrocarbon chains of egg yolk phosphatidylcholine (EggPC)/sodium taurocholate mixed micelles suggested considerably ordered arrangement of the hydrocarbon chains, while isotropic ESR spectra and relatively large fluidity of EggPC/octylglucoside mixed micelles indicated rather disordered arrangement of hydrocarbon chains of EggPC.

An effect of antibiotic amphotericin B on ion transport across model lipid membranes and tonoplast membranes

A pH sensitive fluorescence probe piranine trisulfonate, entrapped inside small unilamellar liposomes formed with egg yolk phosphatidylcholine, was applied to investigate effect of polyene antibiotic amphotericin B (AmB) on proton transport across lipid membranes. Time dependencies of fluorescence-monitored pH changes inside lipid vesicles, upon sudden acidification of the liposome suspension, were analyzed in terms of two-exponential kinetics. It appears that addition of AmB at 3 mol%, with respect to lipid, considerably increases the rate constant of the fast component of proton transport (a change from (60 to 149) × 10 −3 s −1) and decreases the rate constant of the slow component (a change from (11 to 5) × 10 −3 s −1). Incorporation of 0.1 mol% AmB results in the decrease of both parameters (to (33 and 2) × 10 −3 s −1, respectively). The increase in the rate of proton transfer across the lipid membrane is interpreted as related to the formation of membrane channels by AmB, at higher concentration of the drug or nonspecific destabilization of the membrane structure. At low concentrations, at which formation of molecular structures of AmB is not possible, the antibiotic molecules are oriented horizontally with respect to the plane of the membrane and act in making the membrane more compact and less permeable to ions. The presence of sterols (cholesterol, ergosterol and cholesterol dimer) in the lipid phase, in the concentration 3 mol% and lower, decreased the rate constants of proton transfer across the membranes but did not influence significantly the effect of AmB on the ion transport. The presence of AmB in the bathing solutions of tonoplast membranes isolated from Conocephalum conicum at the concentrations range 1 × 10 −7 to 3.6 × 10 −5 does not influence considerably the ion current, as monitored by means of the patch-clamp technique.

Synthesis of phosphatidylcholine: An improved method without using the cadmium chloride complex of sn-glycero-3-phosphocholine

An improved safe method that does not contaminate the environment with cadmium chloride, a toxic heavy metal salt, was developed for the synthesis of phosphatidylcholine (PC). PC was synthesized from sn-glycero-3-phosphocholine (GPC) and fatty acid in one step under mild conditions without the use of cadmium chloride. GPC was prepared from egg yolk PC and adsorbed by kieselguhr in a Teflon vessel. The GPC on kieselguhr was acylated with fatty acid in the presence of two reagents, dicyclohexylcarbodiimide for synthesis of fatty acid anhydride and 4-dimethylaminopyridine as an acylating catalyst, at 30 °C overnight. The PC thus produced was purified by silica gel column chromatography. The yield of dioleoyl PC was 90% based on the starting material, GPC.

Incorporation of carotenoid esters into liposomes

Carotenoid esters are investigated for their interaction with liposomal membranes and compared with their corresponding free (non-esterified) carotenoids. A monoester (β-cryptoxanthin) and two diesters (zeaxanthin and lutein) were chosen. Egg yolk phosphatidylcholine liposomes served as the membrane model. We measured the sizes of the liposomes by photon correlation spectroscopy. The incorporation yields were determined spectrophotometrically. From liposomes simultaneously doped with the fluorescent dye Laurdan, fluidity changes of the liposomes were obtained. In summary, the results indicate that the carotenoid esters: (i) get incorporated, but at a lower yield than their corresponding free carotenoids, (ii) also increase the membrane rigidity as do the free carotenoids, and (iii) increase the liposome sizes significantly, but after extrusion through an 0.1 μm filter the sizes resemble with the exception of the liposomes incorporated with lutein diesters, they remain bigger indicating an elastic property due to two different accessible locations in the membrane.

Biliary cholesterol crystallization characterized by single-crystal cryogenic electron diffraction

Cholesterol crystals are the building blocks of cholesterol gallstones. The exact structure of early-forming crystals is still controversial. We combined cryogenic-temperature transmission electron microscopy with cryogenic-temperature electron diffraction to sequentially study crystal development and structure in nucleating model and native gallbladder biles. The growth and long-term stability of classic cholesterol monohydrate (ChM) crystals in native and model biles was determined. In solutions of model bile with low phospholipid-to-cholesterol ratio, electron diffraction provided direct proof of a novel transient polymorph that had an elongated habit and unit cell parameters differing from those of classic triclinic ChM. This crystal is exactly the monoclinic ChM phase described by Solomonov and coworkers (Biophysical J., In press) in cholesterol monolayers compressed on the air-water interface. We observed no evidence of anhydrous cholesterol crystallization in any of the biles studied. In conclusion, classic ChM is the predominant and stable form in native and model biles. However, under certain (low phospholipid) conditions, transient intermediate polymorphs may form. These findings, documenting single-crystal analysis in bulk solution, provide an experimental approach to investigating factors influencing biliary cholesterol crystal nucleation and growth as well as other processes of nucleation and crystallization in liquid systems.

Nanoparticulate paclitaxel loaded into sterically stabilized mixed phospholipid micelles to improve chemotherapy of breast cancer

Active targeting of water-insoluble chemotherapeutic drugs, such as paclitaxel, to breast cancer is a highly desirable because of its associated increase in anticancer efficacy coupled with reduced systemic drug toxicity. However, rational design of these drug delivery platforms should take into account both pathobiological attributes of breast cancer, such as enhanced permeability and retention phenomenon and overexpression of vasoactive intestinal peptide (VIP) receptors, as well as biophysical properties of its ingredients, including ease of preparation, water insoluble drug loading capacity, steric hindrance, nanosize, and scale-up production and storage. To this end, we developed and tested a novel biocompatible and biodegradable nanoparticulate formulation of VIP-conjugated sterically stabilized phospholipid mixed micelles (SSMM-VIP; size ~ 14 nm) composed of disteraroyl phosphatidylethanolamnine-poly(ethylenglycol-2000) and egg yolk phosphatidylcholine. This construct solubilized 1 mg/ml paclitaxel (P-SSMM-VIP) and retained its biophysical properties upon lyophylization and reconstitution in saline. Moreover, it exhibited a twofold increase in cytotoxicity to MCF-7 breast cancer cells in comparison with P-SSMM and paclitaxel in DMSO (P < 0.05). In addition, the construct targeted VIP receptors overexpressed in methyl nitrosurea (MNU)-induced in situ rat breast cancer tissues. There was a twofold increase in accumulation of intravenously administered P-SSMM-VIP (1 mg/kg) in MNU-induced rat breast cancer, coupled with a significantly greater regression of breast cancer in comparison with P-SSMM and Taxol (P < 0.05). At the same time there was a significant reduction in P-SSMM-VIP accumulation in bone marrow, spleen and other organs in comparison with P-SSMM and Taxol (P < 0.05). There was no significant change in systemic arterial pressure during administration of P-SSMM-VIP. Collectively, these data indicate that actively targeting paclitaxel passively loaded into biocompatible, biodegradable, long-circulating SSMM to breast cancer through VIP receptors improves drug efficacy and reduces its uptake in injury-prone normal tissues. We suggest that P-SSMM-VIP is an efficacious and safe, actively targeted drug delivery platform to treat breast cancer.