- Nano Express
- Open Access
Self-Assembled Polymeric Micellar Nanoparticles as Nanocarriers for Poorly Soluble Anticancer Drug Ethaselen
© to the authors 2009
- Received: 10 June 2009
- Accepted: 19 August 2009
- Published: 16 September 2009
A series of monomethoxy poly(ethylene glycol)-poly(lactide) (mPEG-PLA) diblock copolymers were synthesized, and mPEG-PLA micelle was fabricated and used as a nanocarrier for solubilization and delivery of a promising anticancer drug ethaselen. Ethaselen was efficiently encapsulated into the micelles by the dialysis method, and the solubility of ethaselen in water was remarkably increased up to 82 μg/mL before freeze-drying. The mean diameter of ethaselen-loaded micelles ranged from 51 to 98 nm with a narrow size distribution and depended on the length of PLA block. In vitro hemolysis study indicated that mPEG-PLA copolymers and ethaselen-loaded polymeric micelles had no hemolytic effect on the erythrocyte. The enhanced antitumor efficacy and reduced toxic effect of ethaselen-loaded polymeric micelle when compared with ethaselen-HP-β-CD inclusion were observed at the same dose in H22human liver cancer cell bearing mouse models. These suggested that mPEG-PLA polymeric micelle nanoparticles had great potential as nanocarriers for effective solubilization of poorly soluble ethaselen and further reducing side effects and toxicities of the drug.
- Monomethoxy poly(ethylene glycol)-poly(lactide)
- Polymeric micelles
- Antitumor efficacy
Recently, polymeric micelles as a means to solubilize poorly water-soluble drugs have attracted much attention [4–6]. Generally, block copolymers with concentration above the critical association concentration (CAC) self-assemble into spherical polymeric micelles with a core–shell structure in water: the hydrophobic segments aggregate to form an inner core being able to accommodate hydrophobic drugs with improved solubility by hydrophobic interactions; the hydrophilic shell consists of a brush-like protective corona that stabilizes the micelles in aqueous solution [7–9]. Polymeric micelles as novel drug vehicles present numerous advantages, such as reduced side effects of anticancer drugs, selective targeting, stable storage and prolonged blood circulation time [9, 10]. Furthermore, polymeric micelles possess a nanoscale size range with a narrow distribution, and they can achieve higher accumulation at the target site through an enhanced permeation retention effect (EPR effect) . They can protect drugs against premature degradation in vivo owing to their core–shell architecture [12, 13]. More importantly, polymeric micelles are fabricated according to the physicochemical properties of drugs and the compatibility between the core of micelles and drug molecules [9, 14].
Biodegradable polymers, especially aliphatic polyesters such as polylactide (PLA), poly(DL-lactic-co-glycolic acid) (PLGA) and poly(ε-caprolactone) (PCL) have attracted much attention as biomaterials due to their biocompatibility, degradability and nontoxicity. They have been applied in a wide range of biological systems ranging from drug delivery to tissue engineering. PLA is the most attractive candidate. A number of PLA-based amphiphilic block copolymer micelles have been commonly used for solubilization of hydrophobic drugs [15–17]. In our research, PLA was also chosen as hydrophobic segment of a block copolymer due to the higher compatibility between the micelle core-forming PLA and the ethaselen molecules. In previous work, our group has prepared micellar ethaselen formulation employing PLA-based copolymers and characterized its drug loading contents and stability (data not shown), which have demonstrated the compatibility between the core of micelles and the ethaselen molecules. Polyethylene glycol (PEG) is frequently chosen as a hydrophilic segment to complement a hydrophobic segment due to the outstanding physicochemical and biological properties including solubility in water and in organic solvents, nontoxicity and filterability through kidney when the molar mass is below 30,000 . In addition, PEG is able to form a palisade avoiding protein adsorption and subsequent nonspecific uptake by the reticuloendothelial system (RES) after intravenous injection.
In the present work, amphiphilic diblock copolymers monomethoxy poly(ethylene glycol)-poly(lactide) (mPEG-PLA) with molecular weight of 2,500, 5,000, 10,000 and 15,000 for PLA block were strategically designed and synthesized by ring-opening polymerization ofd,l-dilactide (d,l-LA) in the presence of mPEG with molecular weight of 5,000, respectively. The micelles preparation, ethaselen solubilization and micelles properties were investigated by the UV–vis assay, size measurement and TEM in the micellar solution or in the state of lyophilized powder. The CAC of mPEG-PLA was measured by using the standard fluorescence substance of pyrene. Ethaselen-loaded polymeric micelle was evaluated with respect to its hemolytic toxicity and antitumor efficacy.
d,l-Dilactide was obtained from Fluka. Stannous 2-ethylhexanoate and monomethoxy poly(ethylene glycol) with molecular weight of 5,000 were purchased from Sigma–Aldrich. All other chemicals and reagents were of analytical grade or better and used without further purification.
Male Kunming mice were obtained from Experimental Animal Center of Peking University and acclimatized for 7 days after arrival. New Zealand White rabbits were purchased from the same supplier and acclimatized for 2 days after arrival. All animals were provided with standard food and water ad libitum and were exposed to alternating 12-h periods of light and darkness. Temperature and relative humidity were maintained at 25 °C and 50%, respectively. All care and handling of animals were performed with the approval of Institutional Authority for Laboratory Animal Care of Peking University.
Synthesis and Characterization of mPEG-PLA
mPEG-PLA diblock copolymers were synthesized from d,l-dilactide and mPEG using stannous 2-ethylhexanoateas as a catalyst as described previously  with modification. Briefly, d,l-dilactide was recrystallized in ethyl acetate at room temperature until the racemic mixture melting point was attained (124–126 °C), and mPEG was used without further purification. The synthesized diblock copolymers were referred to as mPEGx-PLAy. x and y represented the weight-averaged molecular weight of the mPEG and PLA block in kDa. mPEG5-PLA2.5, for example, consisted of a 5 kDa mPEG block connected to a 2.5 kDa PLA block.
mPEGx-PLAy was obtained by modulating the feed ratio ofd,l-dilactide and mPEG. The molecular weight of mPEG block was 5,000 (mPEG5). The molecular weights of PLA block were 2,500 (PLA2.5), 5,000 (PLA5), 10,000 (PLA10) and 15,000 (PLA15), respectively. As a typical example, the synthesis of mPEG5-PLA15 was carried out as follows: to remove any trace of water, the starting materials (5 g of mPEG5 and 15 g ofd,l-dilactide) were each dissolved in 100 mL toluene in a round-bottomed flask. About 40 mL of toluene was distilled off using a water separator. The water-free solutions were united in a three-neck flask, a precisely weighed amount of 25 mg stannous 2-ethylhexanoate was added, and the mixture was refluxed for 24 h at 120 °C under nitrogen atmosphere. After toluene was distilled off with a rotary evaporator, the residue was redissolved by the addition of appropriate amount of chloroform, and vigorously stirred and precipitated in diethyl ether at 0 °C, and then filtered. The precipitated polymer was dried in a desiccator at room temperature under high vacuum. After drying for 2 days, white solid powder of the copolymer was obtained. The resulting copolymers were dissolved in CDCl3, and1H-NMR spectra were taken at 300 MHz with trimethylsilane (TMS) as internal reference standard using a Bruker MSL2300 spectrometer (Bruker, Germany). The copolymer molecular weights were determined by gel permeation chromatography (GPC).
Determination of Critical Association Concentration (CAC)
The CAC values of mPEG-PLA diblock copolymers were determined by fluorescence spectroscopy using pyrene (Fluka, >99%) as a hydrophobic probe . Briefly, a known amount of pyrene in acetone was added to each of a series of 50 mL vials, and the acetone was evaporated, then a known amount of various concentrations of mPEG-PLA solutions in acetone were added to each vial, and the acetone was evaporated. The appropriate amount of distilled water was then added to each vial to obtain polymeric aqueous solutions with final concentration of 2.24 × 10−4–224 mg/L. The final concentration of pyrene was 6.0 × 10−7 mol/L. The sample solutions were kept in a constant temperature shaking water bath at 37 °C for 24 h to equilibrate the pyrene and the micelles, and cooled overnight at room temperature. The solutions were filtered with a 0.22 μm pore-sized filtration membrane (Millex-GV, Millipore, USA). Fluorescence spectra of pyrene were recorded with a Shimadzu RF-5301 PC fluorescence spectrometer. The excitation wavelength used was 333 and 335 nm, and the emission spectra were recorded at 390 nm. The peak height intensity ratio (I 335/I 333) of the peak of 335 nm to the peak of 333 nm was plotted against the logarithm of polymer concentration. Two tangents were then drawn, one to the curve at high concentrations and another through the points at low concentrations. The CAC value was taken from the intersection between the two tangents.
Preparation of Ethaselen-Loaded Polymeric Micelles
Particle Size and Morphology Analysis
The average hydrodynamic radius of the micelles was determined by dynamic light scattering (DLS) (Zetasizer ZEN 3500, Malvern, UK). All DLS measurements were done with an angle detection of 173° at 25 °C after diluting the dispersion to an appropriate volume with water. The results were the mean values of three experiments for the same sample. The morphology of micellar nanoparticles was also observed by transmission electron microscopy (TEM) (Hitachi-500, Hitachi, Japan). To improve the contrast, the samples were treated with a 1 wt% phosphotungstic acid solution for 2 h, deposited on copper grids, and allowed to dry for 48 h before TEM examination.
Physical Stability of Ethaselen-Loaded Micelles
The lyophilized powder and the polymeric micelle solution were stored at room temperature. Their physical stability was monitored over time by dynamic light scattering and visually for signs of opalescence and precipitation. The leakage percent was also measured by using the same method as the determination of LC and EE described earlier. In addition, the concentrated magnitude of micelle solution by freeze-drying compared with initially prepared micelle solution was evaluated by comparing the volume of the reconstituted micelle solution of lyophilized powder with that of initially prepared micelle solution. It should be noted that the volume of the reconstituted micelle solution of lyophilized powder was the smallest volume of physiological saline in which the obtained lyophilized powder was redissolved to produce a clear micelle solution.
Assay for Antitumor Efficacy
Sixty of male Kunming mice (body weight = 18–22 g, Peking University Experimental Animal Center, SPF-level, Quality certificated Number: SCXK 2007-0008) were randomly divided into six groups. And seven-day-old liver cancer H22ascites (0.2 mL, 2 × 106cells) were transplanted subcutaneously into the right axilla of each mouse of the groups. The mice were treated as follows: negative control group (normal saline); ethaselen-HP-β-CD group (1 mg/kg body weight); ethaselen-loaded mPEG5-PLA2.5 micelle group (1 mg/kg body weight, low dose); ethaselen-loaded mPEG5-PLA2.5 micelle group (2 mg/kg body weight, middle dose); ethaselen-loaded mPEG5-PLA2.5 micelle group (4 mg/kg body weight, high dose). All the groups were administered through the tail vein of animals once daily for 10 days, starting 24 h after tumor implantation. At day 11, all the mice were killed by cervical dislocation following by separation and measurement of the tumor block. The antitumor efficacies of each formulation were evaluated by tumor inhibition rate, which was calculated by the following formula: inhibition rate = (tumor weight of test group − tumor weight of negative control group)/tumor weight of negative control group × 100%.
All data were expressed as mean ± SD (standard deviation). Comparisons between the group means were evaluated by the unpairedt test. The statistical significance of differences among more than two groups was determined by one-way ANOVA. A value ofp < 0.05 was regarded as significant.
Synthesis and Characterization of mPEG-PLA
Survey on the composition of mPEG-PLAs
Feed ratio mPEG/DL-LA
M w a
M n b
Found ratio mPEG/PLA
Characterization of micelles prepared from a series of mPEG-PLA copolymers
Characterization of mPEG-PLA Micelles
Core–shell type polymeric micelles were prepared by dialysis method. Size and size distribution of micelles were measured by DLS. The mean diameter ranged from 44 to 85 nm for drug-free micelles and from 51 to 98 nm for drug-loaded micelles (Table 2). It appeared that the micelle size gradually increased with increasing in the length of PLA chains. This result was in agreement with the characteristic of amphiphilic copolymer micelles, i.e., the shorter the hydrophobic block length, the smaller the micelles. It might be attributed to the fact that it is difficult to form compact polymeric micelles for amphiphilic copolymers with longer hydrophobic chain length. In addition, similar to drug-loaded micelles reported earlier [15, 27], the size of drug-loaded micelles was about 10 nm bigger than that of drug-free micelles, suggesting that ethaselen molecules were trapped in the hydrophobic inner cores and that these entrapped ethaselen molecules increased the average size of ethaselen-loaded polymeric micelles. Importantly, the PI values of ethaselen-loaded polymeric micelles were fairly low, indicating a monodisperse distribution . Among four polymeric micelles, the size of mPEG5-PLA2.5 micelles was smallest.
Drug loading content (LC, w/w%) and entrapment efficiency (EE, w/w%) of polymeric micelles were calculated by absorbance of ethaselen at 320 nm from UV. The results were also summarized in Table 2. It could be found that mPEG5-PLA2.5 micelles exhibited the highest LC and EE, about 16 and 24%, respectively, while only 1.61% of LC for hydroxylpropyl beta cyclodextrin (HP-β-CD) inclusion . The solubility of ethaselen in water was improved to be about 82 μg/mL before freeze-drying, which was about 32 times higher than that of ethaselen in water. Notably, the results presented in Table 2 were not the general trend that LC and EE of polymeric micelles increased with increasing hydrophobic chain length. This finding may be assigned to the factors contributing to LC and EE. In general, LC and EE depend on the composition of the copolymers, initial diblock copolymeric concentration or the feed weight ratio of the drug to the copolymer, solvent used in formulation process and micelle preparation method and so on. Therefore, the length of hydrophobic chains was not the only one factor influencing LC and EE. When the factors mentioned earlier except the composition of the mPEG-PLA copolymers are optimal for all copolymers, the length of hydrophobic chains is a vital one. In case of our experiment, all micelles were prepared in the same conditions, which may not be optimal for all copolymers, thereby the result was not the general trend. The other possible reason might be filtration step removed larger particles. This phenomenon was also seen in previous report . Nevertheless, the result would not have influence on the subsequent experiments.
Hemolytic Toxicity of Micelles
In vivo Antitumor Efficacy
In vivo antitumor effect of ethaselen-loaded mPEG5-PLA2.5 micelle and ethaselen-HP-β-CD in H22human liver cancer cell bearing mice model ( ,n = 10)
Body weight (g)
Tumor weight (g)
Inhibition rate (%)
20.38 ± 0.94
28.01 ± 2.24
1.29 ± 0.26
19.94 ± 0.86
26.94 ± 2.11*
0.81 ± 0.13*
20.52 ± 0.66
28.10 ± 1.65
0.71 ± 0.12*,**
20.92 ± 0.58
28.75 ± 1.76
0.57 ± 0.17*,**,***
20.20 ± 0.42
28.42 ± 1.81
0.53 ± 0.11*,**,***,****
The body weight of mice treated with physiological saline without drug continuously increased due to probably its nontoxic effect as well as the rapid growth of tumor (Table 3). In ethaselen-loaded polymeric micelle treated groups at all doses, the body weight of mice did not significantly increase. This might be due to their antitumor efficacies thereby the slower growth of tumor. On the other hand, ethaselen-HP-β-CD group appeared to have significant weight loss, resulting from the toxic effect of the drug, indicating that this micelle-based drug delivery system could reduce unwanted side effects of anticancer drugs during cancer therapy.
Overall, ethaselen-loaded polymeric micelle possessed improved antitumor activity and reduced toxic side effects of anticancer drug than ethaselen-HP-β-CD mainly due to the enhanced vascular permeability and EPR effect, and passive targeting function although they do not have active targeting function [41–43]. Furthermore, tumor tissues are characterized with leaky blood vessels and the premature lymphatic drainage . Resultantly, we speculated that ethaselen-loaded polymeric micelles would also be a superior formulation for other tumor models.
Nevertheless, ethaselen, as a poorly water-soluble drug, might be physically incorporated into the inner core of the polymeric micelles by hydrophobic interactions. Further, it may avoid RES recognition due to a size smaller than 200 nm. Therefore, it is advantageous for ethaselen to be efficiently encapsulated in micelles. In case of our experiment, the EE was unfavorable, thereby the enhancing of hydrophobic interaction between ethaselen and the inner core of the polymeric micelles would be vital by chemical modification the hydrophobic block of mPEG-PLA.
We have successfully synthesized a series of mPEG-PLA copolymers. It was found that monodispersed micelles self-assembled from mPEG-PLA could effectively solubilize the anticancer drug ethaselen when compared with HP-β-CD inclusion. The hemolysis assay indicated that ethaselen-loaded mPEG-PLA micelles could be administered intravenously at a wide range of drug concentrations. In mice, ethaselen-loaded polymeric micelles showed noticeable antitumor efficacy, and reduced the toxic effect of the drug, compared with ethaselen-HP-β-CD inclusion. These results suggested that polymeric micelles might be an effective drug delivery system for ethaselen for cancer chemotherapy. Nevertheless, the micelles could still be improved, especially with respect to enhancing their entrapment efficiency by modification of inner core of micelles, which are in progress. Better drug retention in the micelle core is a key to ensure prolonged circulation time and eventually maximize drug accumulation at the tumor site via the enhanced permeation and retention effect.
The authors wish to thank Prof. Huihui Zeng from Department of Chemical Biology, Peking University for friendly providing ethaselen.
- Shi CJ, Yu LZ, Yang FG, Zeng HH: Biochem. Biophy. Res. Commun.. 2003, 309: 578. COI number [1:CAS:528:DC%2BD3sXntVGmtrw%3D] COI number [1:CAS:528:DC%2BD3sXntVGmtrw%3D] 10.1016/j.bbrc.2003.08.032View ArticleGoogle Scholar
- Yan J, Deng SJ, Zeng HH: J. Chin. Pharm. Sci.. 2004, 13: 199. COI number [1:CAS:528:DC%2BD2cXps1Kgu7g%3D] COI number [1:CAS:528:DC%2BD2cXps1Kgu7g%3D]Google Scholar
- Cui HM, Zhang CG, Wu FL: Chin. Pharm. Sci.. 2007, 42: 765. COI number [1:CAS:528:DC%2BD1cXntFCgsbg%3D] COI number [1:CAS:528:DC%2BD1cXntFCgsbg%3D]Google Scholar
- Torchilin VP: Adv. Drug Deliv. Rev.. 2002, 54: 235. COI number [1:CAS:528:DC%2BD38XhvVWnsr0%3D] COI number [1:CAS:528:DC%2BD38XhvVWnsr0%3D] 10.1016/S0169-409X(02)00019-4View ArticleGoogle Scholar
- Rösler A, Vandermeulen GW, Klok HA: Adv. Drug Deliv. Rev.. 2001, 53: 95. 10.1016/S0169-409X(01)00222-8View ArticleGoogle Scholar
- Jones M, Leroux J: Eur. J. Pharm. Biopharm.. 1999, 48: 101. COI number [1:CAS:528:DyaK1MXmtVymurk%3D] COI number [1:CAS:528:DyaK1MXmtVymurk%3D] 10.1016/S0939-6411(99)00039-9View ArticleGoogle Scholar
- Shuai X, Merdan T, Schaper AK, Xi F, Kissel T: Bioconjug. Chem.. 2004, 15: 441. COI number [1:CAS:528:DC%2BD2cXjsFGgsrk%3D] COI number [1:CAS:528:DC%2BD2cXjsFGgsrk%3D] 10.1021/bc034113uView ArticleGoogle Scholar
- Riley T, Heald CR, Stolnik S, Garnett MC, Illum L, Davis SS: Langmuir. 2003, 19: 8428. COI number [1:CAS:528:DC%2BD3sXmsFOisLo%3D] COI number [1:CAS:528:DC%2BD3sXmsFOisLo%3D] 10.1021/la020911hView ArticleGoogle Scholar
- Liu L, Li C, Li X, Yuan Z, An Y, He B: J. Appl. Polym. Sci.. 2001, 80: 1976. COI number [1:CAS:528:DC%2BD3MXisVCku7g%3D] COI number [1:CAS:528:DC%2BD3MXisVCku7g%3D] 10.1002/app.1295View ArticleGoogle Scholar
- Pierri E, Avgoustakis K: J. Biomed. Mater. Res. A. 2005, 75: 639. COI number [1:STN:280:DC%2BD2MrmtVWrsg%3D%3D] COI number [1:STN:280:DC%2BD2MrmtVWrsg%3D%3D]View ArticleGoogle Scholar
- Kataoka K, Harada A, Nagasaki Y: Adv. Drug Deliv. Rev.. 2001, 47: 113. COI number [1:CAS:528:DC%2BD3MXhvVajs78%3D] COI number [1:CAS:528:DC%2BD3MXhvVajs78%3D] 10.1016/S0169-409X(00)00124-1View ArticleGoogle Scholar
- Wilhelm M, Zhao CL, Wang YC, Xu RL, Winnik MA, Mura JL, et al.: Macromolecules. 1991, 24: 1033. COI number [1:CAS:528:DyaK3MXhtVehtb8%3D]; Bibcode number [1991MaMol..24.1033W] COI number [1:CAS:528:DyaK3MXhtVehtb8%3D]; Bibcode number [1991MaMol..24.1033W] 10.1021/ma00005a010View ArticleGoogle Scholar
- Barreiro-Iglesias R, Bromberg L, Temchenko M, Hatton TA, Concheiro A, Alvarez-Lorenzo C: J. Control. Release. 2004, 97: 537. COI number [1:CAS:528:DC%2BD2cXkvF2itb0%3D] COI number [1:CAS:528:DC%2BD2cXkvF2itb0%3D]View ArticleGoogle Scholar
- Li PZ, Li XR, Zhou HX, Zhang YH, Wang F, Liu Y: Chin. J. New Drug. 2009, 18: 262. COI number [1:CAS:528:DC%2BD1MXkt12mtr8%3D] COI number [1:CAS:528:DC%2BD1MXkt12mtr8%3D]Google Scholar
- Zhang Y, Jin T, Zhuo RX: Colloids Surf. B: Biointerfaces. 2005, 44: 104. COI number [1:CAS:528:DC%2BD2MXntVCqur0%3D] COI number [1:CAS:528:DC%2BD2MXntVCqur0%3D] 10.1016/j.colsurfb.2005.06.004View ArticleGoogle Scholar
- Lee ES, Na K, Bae YH: J. Control. Release. 2003, 91: 103. COI number [1:CAS:528:DC%2BD3sXmsVSkur4%3D] COI number [1:CAS:528:DC%2BD3sXmsVSkur4%3D] 10.1016/S0168-3659(03)00239-6View ArticleGoogle Scholar
- Le Garrec D, Gori S, Luo L, Lessard D, Smith DC, Yessine MA, et al.: J. Control. Release. 2004, 99: 83. COI number [1:CAS:528:DC%2BD2cXnt1Kjt7w%3D] COI number [1:CAS:528:DC%2BD2cXnt1Kjt7w%3D] 10.1016/j.jconrel.2004.06.018View ArticleGoogle Scholar
- Hu Y, Jiang X, Ding Y, Zhang L, Yang C, Zhang J, et al.: Biomaterials. 2003, 24: 2395. COI number [1:CAS:528:DC%2BD3sXivVCqsb0%3D] COI number [1:CAS:528:DC%2BD3sXivVCqsb0%3D] 10.1016/S0142-9612(03)00021-8View ArticleGoogle Scholar
- Lucke A, Tessmar J, Schnell E, Schmeer G, Göpferich A: Biomaterials. 2000, 21: 2361. COI number [1:CAS:528:DC%2BD3cXmslKmt78%3D] COI number [1:CAS:528:DC%2BD3cXmslKmt78%3D] 10.1016/S0142-9612(00)00103-4View ArticleGoogle Scholar
- Yu BG, Okano T, Kataoka K, Kwon G: J. Control. Release. 1998, 53: 131. COI number [1:CAS:528:DyaK1cXitVCjs78%3D] COI number [1:CAS:528:DyaK1cXitVCjs78%3D] 10.1016/S0168-3659(97)00245-9View ArticleGoogle Scholar
- Zhu S, Qian F, Zhang Y, Tang C, Yin C: Eur. Polym. J.. 2007, 43: 2244. COI number [1:CAS:528:DC%2BD2sXmt1Kis7Y%3D] COI number [1:CAS:528:DC%2BD2sXmt1Kis7Y%3D] 10.1016/j.eurpolymj.2007.03.042View ArticleGoogle Scholar
- Kwon GS, Kataoka K: Adv. Drug Deliv. Rev.. 1995, 16: 295. COI number [1:CAS:528:DyaK2MXpt1amt70%3D] COI number [1:CAS:528:DyaK2MXpt1amt70%3D] 10.1016/0169-409X(95)00031-2View ArticleGoogle Scholar
- Kim SY, Shin IG, Lee YM: J. Control. Release. 1998, 56: 197. COI number [1:CAS:528:DyaK1cXntVGhurk%3D] COI number [1:CAS:528:DyaK1cXntVGhurk%3D] 10.1016/S0168-3659(98)00083-2View ArticleGoogle Scholar
- Li S, Vert M: Macromolecules. 2003, 36: 8008. COI number [1:CAS:528:DC%2BD3sXnsFyqtbc%3D]; Bibcode number [2003MaMol..36.8008L] COI number [1:CAS:528:DC%2BD3sXnsFyqtbc%3D]; Bibcode number [2003MaMol..36.8008L] 10.1021/ma034734iView ArticleGoogle Scholar
- Zhang X, Jackson JK, Burt HM: Inter. J. Pharm.. 1996, 132: 195. COI number [1:CAS:528:DyaK28XjtFSktrc%3D] COI number [1:CAS:528:DyaK28XjtFSktrc%3D] 10.1016/0378-5173(95)04386-1View ArticleGoogle Scholar
- Jeong B, Han Bae Y, Wan Kim S: Colloids Surf. B: Biointerfaces. 1999, 16: 185. COI number [1:CAS:528:DyaK1MXnvFylurk%3D] COI number [1:CAS:528:DyaK1MXnvFylurk%3D] 10.1016/S0927-7765(99)00069-7View ArticleGoogle Scholar
- Yang KW, Li XR, Yang ZL, Li PZ, Wang F, Liu Y: J. Biomed. Mater. Res. Part A. 2009, 87A: 140. COI number [1:CAS:528:DC%2BD1cXhsV2lsr%2FP] COI number [1:CAS:528:DC%2BD1cXhsV2lsr%2FP] 10.1002/jbm.a.31866View ArticleGoogle Scholar
- Harada A, Kataoka K: Macromolecules. 1995, 28: 5294. COI number [1:CAS:528:DyaK2MXmsFejtrY%3D]; Bibcode number [1995MaMol..28.5294H] COI number [1:CAS:528:DyaK2MXmsFejtrY%3D]; Bibcode number [1995MaMol..28.5294H] 10.1021/ma00119a019View ArticleGoogle Scholar
- Chang YC, Chu IM: Eur. Polym. J.. 2008, 44: 3922. COI number [1:CAS:528:DC%2BD1cXhsVKht7zM] COI number [1:CAS:528:DC%2BD1cXhsVKht7zM] 10.1016/j.eurpolymj.2008.09.021View ArticleGoogle Scholar
- Huh KM, Lee SC, Cho YW, Lee J, Jeong JH, Park K: J. Control. Release. 2005, 101: 59. COI number [1:CAS:528:DC%2BD2cXhtVKgu7nE] COI number [1:CAS:528:DC%2BD2cXhtVKgu7nE] 10.1016/j.jconrel.2004.07.003View ArticleGoogle Scholar
- Burt HM, Zhang X, Toleikis P, Embree L, Hunter WL: Colloids Surf. B: Biointerfaces. 1999, 16: 161. COI number [1:CAS:528:DyaK1MXnvFylurs%3D] COI number [1:CAS:528:DyaK1MXnvFylurs%3D] 10.1016/S0927-7765(99)00067-3View ArticleGoogle Scholar
- Zastre J, Jackson J, Bajwa M, Liggins R, Iqbal F, Burt H: Eur. J. Pharm. Biopharm.. 2002, 54: 299. COI number [1:CAS:528:DC%2BD38XoslGgur4%3D] COI number [1:CAS:528:DC%2BD38XoslGgur4%3D] 10.1016/S0939-6411(02)00119-4View ArticleGoogle Scholar
- Savic R, Luo L, Eisenberg A, Maysinger D: Science. 2003, 300: 615. COI number [1:CAS:528:DC%2BD3sXjtVymur8%3D]; Bibcode number [2003Sci...300..615S] COI number [1:CAS:528:DC%2BD3sXjtVymur8%3D]; Bibcode number [2003Sci...300..615S] 10.1126/science.1078192View ArticleGoogle Scholar
- Pratten MK, Lloyd JB, Hörpel G, Ringsdorf H: Die Makromol. Chem.. 1985, 186: 725. COI number [1:CAS:528:DyaL2MXksFWksrY%3D] COI number [1:CAS:528:DyaL2MXksFWksrY%3D] 10.1002/macp.1985.021860406View ArticleGoogle Scholar
- Shiah JG, Dvorák M, Kopecková P, Sun Y, Peterson CM, Kopecek J: Eur. J. Cancer. 2001, 37: 131. COI number [1:CAS:528:DC%2BD3MXntlOmsw%3D%3D] COI number [1:CAS:528:DC%2BD3MXntlOmsw%3D%3D] 10.1016/S0959-8049(00)00374-9View ArticleGoogle Scholar
- Shiah JJ, Sun Y, Peterson CM, Kopecek J: J. Control. Release. 1999, 61: 145. COI number [1:CAS:528:DyaK1MXlsFOrsr8%3D] COI number [1:CAS:528:DyaK1MXlsFOrsr8%3D] 10.1016/S0168-3659(99)00113-3View ArticleGoogle Scholar
- Yokoyama M, Fukushima S, Uehara R, Okamoto K, Kataoka K, Sakurai Y, et al.: J. Control. Release. 1998, 50: 79. COI number [1:CAS:528:DyaK2sXnvFSgu78%3D] COI number [1:CAS:528:DyaK2sXnvFSgu78%3D] 10.1016/S0168-3659(97)00115-6View ArticleGoogle Scholar
- Zuo Z, Tam YK, Diakur J, Wiebe LI: J. Pharm. Pharm. Sci.. 2002, 5: 292. COI number [1:CAS:528:DC%2BD3sXivV2itro%3D] COI number [1:CAS:528:DC%2BD3sXivV2itro%3D]Google Scholar
- Frijlink HW, Franssen EJ, Eissens AC, Oosting R, Lerk CF, Meijer DK: Pharm. Res.. 1991, 8: 380. COI number [1:CAS:528:DyaK3MXitFChtbY%3D] COI number [1:CAS:528:DyaK3MXitFChtbY%3D] 10.1023/A:1015857902238View ArticleGoogle Scholar
- Piel G, Evrard B, Van Hees T, Delattre L: Int. J. Pharm.. 1999, 180: 41. COI number [1:CAS:528:DyaK1MXhvFCru7Y%3D] COI number [1:CAS:528:DyaK1MXhvFCru7Y%3D] 10.1016/S0378-5173(98)00403-7View ArticleGoogle Scholar
- Jeong Y, Na HS, Cho KO, Lee HC, Nah JW, Cho CS: Int. J. Pharm.. 2009, 365: 150. COI number [1:CAS:528:DC%2BD1cXhsVKnur7J] COI number [1:CAS:528:DC%2BD1cXhsVKnur7J] 10.1016/j.ijpharm.2008.08.011View ArticleGoogle Scholar
- Greish K, Nagamitsu A, Fang J, Maeda H: Bioconjug. Chem.. 2005, 16: 230. COI number [1:CAS:528:DC%2BD2cXhtVWhsbrN] COI number [1:CAS:528:DC%2BD2cXhtVWhsbrN] 10.1021/bc040297gView ArticleGoogle Scholar
- Maeda H, Wu J, Sawa T, Matsumura Y, Hori K: J. Control. Release. 2000, 65: 271. COI number [1:CAS:528:DC%2BD3cXhtl2qsLo%3D] COI number [1:CAS:528:DC%2BD3cXhtl2qsLo%3D] 10.1016/S0168-3659(99)00248-5View ArticleGoogle Scholar
- Bae Y, Kataoka K: Adv. Drug Deliv. Rev.. 2009, 61: 768. COI number [1:CAS:528:DC%2BD1MXnsl2gtb8%3D] COI number [1:CAS:528:DC%2BD1MXnsl2gtb8%3D] 10.1016/j.addr.2009.04.016View ArticleGoogle Scholar