Preparation and characterization of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) microspheres for controlled release of buprofezin.

Extensive application of pesticides has caused a lot of environmental pollution and health problems, prompting the development of highly efficient and lowly toxic pesticide formulations. Here, buprofezin (BPF)-loaded poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-HH)) microspheres were prepared by O/W emulsion/solvent evaporation method. Under optimal conditions (P(HB-HH) 7.07% (w/v) and PVA 1.84% (w/v)), the spherical and monodispersed microspheres were obtained. The average particle size, pesticide loading (PL), and encapsulation efficiency (EE) of the optimized microspheres were 1.2 µm, 15.68%, and 78%, respectively. Release of 80% BPF from the microspheres in pH 5 (192 h) was faster than that in pH 7 (228 h) and 8 (204 h). Moreover, kinetic analysis indicated that BPF release behaved in a non-Fickian diffusion manner (0.43 < n < 0.85) and the release mechanism was the combined effects of pesticide diffusion and hydrolysis of polymer. The bioassay and toxicity results showed that encapsulation of BPF could exhibit high efficacy on the target organism and low toxicity to the non-target organism. Therefore, these results demonstrated that BPF-loaded P(HB-HH) microspheres with good stability were prepared successfully, and they could be further explored for constructing other highly efficient and lowly toxic pesticide formulations.

Folate-decorated poly(3-hydroxybutyrate-co-3-hydroxyoctanoate) nanoparticles for targeting delivery: optimization and in vivo antitumor activity.

CONTEXT: Doxorubicin (DOX)-loaded folate-targeted poly(3-hydroxybutyrate-co-3-hydroxyoctanoate) [P(HB-HO)] nanoparticles [DOX/FA-PEG-P(HB-HO) NPs] have potential application in clinical treatments for cervical cancer due to specific affinity of folate and folate receptor in HeLa cells. OBJECTIVE: The aim of this study was to develop an optimized formulation for DOX/FA-PEG-P(HB-HO) NPs, and investigate the targeting and efficacies of the nanoparticles. MATERIALS AND METHODS: DOX/FA-PEG-P(HB-HO) NPs were prepared by W1/O/W2 solvent extraction/evaporation method, and an orthogonal experimental design [L9 (3(4))] was applied to establish the optimum conditions. The physico-chemical characteristics, microscopic observation and in vivo antitumor study of the nanoparticles were evaluated. RESULTS: The optimum formulation was obtained with DOX 10% (w/v), FA-PEG-P(HB-HO) 6.5% (w/v), PVA 3%(w/v) and oil phase/internal water phase volume ratio of 3/1. The size distribution, drug loading and encapsulation efficiency of the optimized nanoparticles were 150-350 nm, 29.6 ± 2.9% and 83.5 ± 5.7%, respectively. In vitro release study demonstrated that 80% of the drug could release from the nanoparticles within 11 days. Furthermore, in vitro microscopic observation and in vivo antitumor study showed that DOX/FA-PEG-P(HB-HO) NPs could inhibit HeLa cells effectively, and the tumor inhibition rate (TIR) in vivo was 76.91%. DISCUSSION AND CONCLUSIONS: DOX/FA-PEG-P(HB-HO) NPs have been successfully developed and optimized. In vitro drug release study suggested a sustained release profile. Moreover, DOX/FA-PEG-P(HB-HO) NPs could effectively inhibit HeLa cells with satisfying targeting, and reduce side effects and toxicity to normal tissues. DOX/FA-PEG-P(HB-HO) NPs were superior in terms of inhibiting HeLa tumor over non-targeted formulations therapy.

Quantification of DOX bioavailability in biological samples of mice by sensitive and precise HPLC assay.

CONTEXT: Doxorubicin (DOX)-loaded folate-targeted poly(3-hydroxybutyrate-co-3-hydroxyoctanoate) [P(HB-HO)] nanoparticles [DOX/FA-PEG-P(HB-HO) NPs] were prepared by the W1/O/W2 solvent extraction/evaporation method for applications in cancer treatment. However, the biodistribution, pharmacokinetics, and targeting of the nanoparticles (NPs) have not yet been studied. OBJECTIVE: The biodistribution, pharmacokinetics, and targeting of DOX/FA-PEG-P(HB-HO) NPs were evaluated in female BALB/c nude mice bearing HeLa tumors. MATERIALS AND METHODS: Three DOX formulations were injected into the tail vein of the mice at a dosage of 5 mg/kg. At each time point, blood and various tissues were collected. All samples were then processed and analyzed by a validated high performance liquid chromatographic (HPLC) method. RESULTS: The t1/2 values of DOX/P(HB-HO) NPs and DOX/FA-PEG-P(HB-HO) NPs were 2.7- and 3.5-times higher than that of free DOX. No significant difference (p > 0.05) was found in Cmax between the NPs and free DOX. The Tmax values of the two NPs were prolonged from 0.25 to 1 h. The AUC0-t values were 1.55- and 3.05-folds higher than that of free DOX, and MRT increased to 15.99 h for DOX/P(HB-HO) NPs and 25.14 h for DOX/FA-PEG-P(HB-HO) NPs. For DOX/FA-PEG-P(HB-HO) NPs, the DOX content in the tumors were 10.81- and 3.33-times higher than those for free DOX and DOX/P(HB-HO) NPs at 48 h, respectively. DISCUSSION AND CONCLUSIONS: DOX/FA-PEG-P(HB-HO) NPs displayed reduced cardiac toxicity and improved bioavailability. Moreover, the NPs exhibited a significant extent of DOX accumulation in the tumors, thus suggesting that folate-targeted NPs could effectively transport into HeLa tumors with satisfying targeting.

Isolation and antitumor efficacy evaluation of a polysaccharide from Nostoc commune Vauch.

Nostoc commune Vauch. has been traditionally used as a healthy food and medicine for centuries especially in China. It has been demonstrated that the polysaccharides isolated from Nostoc commune Vauch. exhibit strong antimicrobial and antioxidant activities. However, little is known about their anticancer activities and the underlying mechanisms of action. Herein, we report the isolation of a polysaccharide from Nostoc commune Vauch. (NVPS), and its physicochemical properties were analyzed. In an attempt to demonstrate the potential application of NVPS in tumor chemotherapy, the in vitro antitumor activity was determined. NVPS significantly suppressed the growth and proliferation of MCF-7 and DLD1 cells. The molecular mechanism underlying this in vitro antitumor efficacy was elucidated, and the results indicated that NVPS simultaneously triggered intrinsic, extrinsic and endoplasmic reticulum stress (ERS)-mediated apoptotic signaling pathways. Collectively, these findings demonstrate that NVPS could be used as a novel promising source of natural antitumor agents.

Water stress proteins from Nostoc commune Vauch. exhibit anti-colon cancer activities in vitro and in vivo.

Nostoc commune has been traditionally used in China as a health food and medicine. The water stress proteins (WSP) of Nostoc commune are the major component of the extracellular matrix. This study purified and identified the water stress proteins (WSP1) from Nostoc commune Vauch., which could inhibit the proliferation of human colon cancer cell lines. The IC50 values of WSP1 against DLD1, HCT116, HT29, and SW480 cells were 0.19 ± 0.02, 0.21 ± 0.03, 0.39 ± 0.05, and 0.41 ± 0.01 µg/µL, respectively. Notably, it displayed very little effect on the normal human intestinal epithelial FHC cell line. The IC50 value of WSP1 against FHC cells was 0.67 ± 0.05 µg/µL. Moreover, the growth of DLD1 xenografted tumors in nude mice were significantly suppressed in the WSP1 treated group. Mechanistically, the cell-cycle analysis revealed that WSP1 induced growth inhibition by G1/S arrest. Meanwhile, Western blotting and immunohistochemistry assays showed WSP1 could activate caspase-8, -9, and -3, along with subsequent PARP cleavage. Furthermore, the pan-caspase inhibitor, z-VAD-FMK, partly reversed the effect caused by WSP1, confirming that WSP1 induced cell apoptosis through caspase-dependent pathway. Collectively, WSP1 has targeted inhibition for colon cancer proliferation both in vitro and in vivo and it is valuable for future exploitation and utilization as an antitumor agent.

Preparation and biological evaluation of a novel selenium-containing exopolysaccharide from Rhizobium sp. N613.

In order to obtain a low toxic antitumor agent and an organic selenium source, an exopolysaccharide obtained from Rhizobium sp. N613 (REPS) was modified by selenious acid using barium chloride as the catalyst. The reaction conditions were optimized by response surface methodology (RSM), and the optimal conditions for preparation of selenium-containing REPS (Se-REPS) were obtained. The selenium content of Se-REPS was 790 µg/g under these conditions. The molecular structure of Se-REPS was confirmed by FTIR. In vitro antitumor activity of Se-REPS was evaluated by MTT assay, and the results indicated that Se-REPS could significantly inhibit the growth of S180 and HepG2 cells. Furthermore, Se-REPS exhibited comparable in vivo antitumor efficacy to cyclophosphamide at same concentrations. In addition, Se-REPS could substantially elevate the thymus and spleen indices in tumor-bearing mice. This study demonstrates that Se-REPS holds great potential to be a desirable antitumor agent for therapeutic and immunomodulatory applications.

Preparation and characterization of novel microparticles based on poly(3-hydroxybutyrate-co-3-hydroxyoctanoate).

In this study, a novel poly(3-hydroxybutyrate-co-3-hydroxyoctanoate) (P(HB-HO)) microparticle with an encapsulated antibiotic (azithromycin, AZI) was prepared by the electrospinning method. The resulting microparticles were evaluated for surface morphology, particle size, drug loading, encapsulation efficiency, in vitro drug-release and degradation. The in vitro cytotoxicity and in vivo pharmacokinetics were also studied. The sizes of microparticles showed a narrow monodisperse size distribution approximately from 3 to 30 µm. In vitro release experiments exhibited sustained release behavior. The results of in vitro degradation tests demonstrated that the mass loss of the P(HB-HO) microparticles was 9.6% and the morphology varied greatly within 24 weeks. P(HB-HO) showed no cytotoxicity to fibroblast when incubated with blank P(HB-HO) microparticles during the tests. The in vivo pharmacokinetic study demonstrated that the microparticles exhibited longer circulation properties than free AZI. It is suggested that novel AZI-loaded P(HB-HO) microparticles can be utilized as a biodegradable and biocompatible drug delivery system.

Folate-mediated poly(3-hydroxybutyrate-co-3-hydroxyoctanoate) nanoparticles for targeting drug delivery.

A novel targeting drug delivery system (TDDS) has been developed. Such a TDDS was prepared by W(1)/O/W(2) solvent extraction/evaporation method, adopting poly(3-hydroxybutyrate-co-3-hydroxyoctanoate) [P(HB-HO)] as the drug carrier, folic acid (FA) as the targeting ligand, and doxorubicin (DOX) as the model anticancer drug. The average size, drug loading capacity and encapsulation efficiency of the prepared DOX-loaded, folate-mediated P(HB-HO) nanoparticles (DOX/FA-PEG-P(HB-HO) NPs) were found to be around 240 nm, 29.6% and 83.5%. The in vitro release profile displayed that nearly 50% DOX was released in the first 5 days. The intracellular uptake tests of the nanoparticles (NPs) in vitro showed that the DOX/FA-PEG-P(HB-HO) NPs were more efficiently taken up by HeLa cells compared to non-folate-mediated P(HB-HO) NPs. In addition, DOX/FA-PEG-P(HB-HO) NPs (IC(50)=0.87 microM) showed greater cytotoxicity to HeLa cells than other treated groups. In vivo anti-tumor activity of the DOX/FA-PEG-P(HB-HO) NPs showed a much better therapeutic efficacy in inhibiting tumor growth, and the final mean tumor volume was 178.91+/-17.43 mm(3), significantly smaller than normal saline control group (542.58+/-45.19 mm(3)). All these results have illustrated that our techniques for the preparing of DOX/FA-PEG-P(HB-HO) NPs developed in present work are feasible and these NPs are effective in selective delivery of anticancer drug to the folate receptor-overexpressed cancer cells. The new TDDS may be a competent candidate in application in targeting treatment of cancers.

Studies on the chemical structure and antitumor activity of an exopolysaccharide from Rhizobium sp. N613.

The molecular structure of the rhizobium exopolysaccharide (REPS) was analyzed by enzymolysis, periodate oxidation, and Smith degradation, and by IR and NMR spectroscopy. The results indicated that REPS was a beta-glucan with a backbone of beta-D-(1-->4)-linked glucose residues and branches of beta-D-(1-->6)-linked glucose residues. The branch was attached to the main chain at the 6-O-position. The molar ratio of 1-->4 and 1-->6 was 2:1. The terminal C3 of the (1-->6)-Glc branch had an O-acetyl group. The molecular weight was estimated to be 35 kDa by Sephadex G-100 column chromatography. The antitumor activity of REPS was evaluated in mice bearing sarcoma 180, hepatoma 22, and Ehrlich ascites carcinoma tumor, respectively. At doses of 10-60 mg/kg, it was observed that tumor formation decreased significantly (P <0.01), but the relative spleen and thymus weight, the phagocytic function of monocytes, lymphocyte proliferation, and serum hemolysis antibody increased significantly (P <0.05). Results of these studies demonstrated that the REPS polysaccharide possessed antitumor activity.