Physical characteristics of nanoliposomes prepared from hydrolyzed cannabis protein

The present study was conducted on the protein extracted from the pulp that obtained by cannabis oil-pressingand loading the bioactive peptides into nanoliposomes. Physical properties of empty and loaded nanoliposemes(average particle size, polydispersity index, and zeta potential) and encapsulation efficiency were evaluated.Moreover, the effect of storage condition (fridge and ambient temperature) on the physical stability and maintainingthe encapsulation efficiency in nanoliposomes loaded with peptides was investigated. Eventually, the effect ofpeptide loading on the nanoparticle chemical structure Fourier-transform infrared (FTIR) as well as the morphologyof nanocarriers scanning electron microscopy (SEM) was evaluated. Physical properties of nanoliposomes wereaffected by the hydrolyzed type. The average particle size and polydispersity index of nanoliposomes varied from79.5 to 101.5 nm and 0.234 to 0.326, depending on the type of loaded peptides. The zeta potential of nanoliposomeswas changed from −10.32 to −15.33 mV when loaded with the peptide obtained by enzymatic hydrolysis during300 min. The efficiency of nanoliposomal encapsulation varied from 90.7 to 81.4%. Thus, peptides with the highestdegree of hydrolysis had the least encapsulation efficiency. The evaluation of physical stability and the maintenanceof encapsulation efficiency indicated the highest stability of nanoliposomes, which were stored at fridge temperature.FTIR spectroscopy in empty nanoliposomes and those loaded with peptides implied the presence of peptides inthe polar regions of phosphatidylcholine, such as the internal regions of the liposomes and the formation of ioniccomplexes between them. Conversely, SEM images of the structural and superficial properties of nanoliposomesindicated the existence of dense and compact clusters of the spherical nanoparticles with flat surfaces.

Evaluation of the internal therapeutic effectiveness of 131I-antiEGFR-BSA-PCL in nude mice with colorectal cancer / 中华放射医学与防护杂志

Objective To investigate the biological effects of internal radiation and therapeutic effectiveness of 131I-labeled anti-epidermal growth factor receptor (EGFR) in colorectal cancer of model mice.Methods Nano-liposome characterized for EGFR-targeting was constructed.The efficacy of cellular binding and uptake of the liposome was evaluated by the analysis of confocal microscopy observation and the iodide uptake assay.After intra-tumor injections of 74 MBq (740 MBq/ml) 131 I-antiEGFR-BSA-PCL,131 I-BSA-PCL,131 I or an equivalent volume of normal saline.The biological effects of internal irradiation and therapeutic efficacy of the liposomes on colorectal cancer modeled in a male BALB/c mouse were evaluated by means of tumor size,body weight,histopathology,and SPECT imaging.Results The confocal fluorescence images showed that the antiEGFR-BSA-PCL was successfully internalized into LS180 cells.The 131I uptake efficacy of 131I-antiEGFR-BSA-PCL was significantly higher than that of 131I-BSA-PCL in LS180 cells (t =2.77-5.40,P ＜ 0.01).Tumor size measurement showed that tumor growth was inhibited by the treatment with 131 I-EGFR-BSA-PCL and 131I-BSA-PCL,but had no significant differences between these two groups (P ＞0.05).It was found that the 131I-antiEGFR-BSA-PCL was markedly taken up by the tumor and reac hed its uptake value of (21.61 ± 1.0 1) and (20.58 ± 0.65)％ ID/g at 72 h following drug injection,which was higher than the uptake value of 131 I (t =9.36,8.69,P ＜ 0.01).SPECT imaging assay showed that,after being injected into mouse tumor,the 131 I-EGFR-BSA-PCL and 131I-BSA-PCL were uniformly distributed inside the tumor.Conclusions 131 I-antiEGFR-BSA-PCL obviously suppresses the development of colorectal cancer in mice.

Study on Targeting of Folic Acid-modified Docetaxel Nano-liposome to Hepatocellular Carcinoma Bel-7402 Cells in vivo and in vitro / 中国药房

OBJECTIVE:To study the targeting of folic acid(FA)-modified docetaxel(DOC)nano-liposome(L-DOC-FA)to hepatocellular carcinoma Bel-7402 cells in vivo and in vitro. METHODS:The cell viability and survival rate of Bel-7402 cells was tested by CCK-8 kit after treated with 0,1,2,5,10 and 20 μg/ml DOC,L-DOC and L-DOC-FA for 24 h. And then,the fluores-cein isothiocyanate was used to label L-DOC and L-DOC-FA nano-liposome,and the rate of L-DOC and L-DOC-FA absorbed by hepatocellular carcinoma Bel-7402 cells were detected. 125I was used to label L-DOC and L-DOC-FA nano-liposome,and then the contents of them in the subcutaneous tumor tissues were detected. 28 Balb/c naked mice were selected and given liver cell suspen-sion via back ih to induce tumor model. After modeling,naked mice were divided into blank control group(normal saline),DOC group(3 mg/kg),L-DOC(3 mg/kg,by DOC)and L-DOC-FA(3 mg/kg,by DOC). They were given relevant medicine intrave-nously once a day for consecutive 30 d. The relative tumor volume in naked mice was detected. RESULTS:DOC,L-DOC and L-DOC-FA all inhibited the cell viability of Bel-7402 cells,the survival rate of cells decreased in concentration-dependant manner;compared with DOC and L-DOC,the cell viability decreased after treated with L-DOC-FA,the survival rate of cells decreased (PL-DOC (31.2%),with statistical significance (P<0.01). The content of L-DOC-FA in tumor was significantly more than that of L-DOC (P<0.01). In addition,3 mg/kg L-DOC-FA showed better inhibitory effect than 3 mg/kg L-DOC and DOC on tumor,and the rela-tive tumor volume was smaller(P<0.01). CONCLUSIONS:L-DOC-FA has obvious targeting to Bel-7402 cells in vivo and in vi-tro,and shows good inhibitory effect on tumor in vivo and in vitro.

Advances in the nanotechnology-based drug delivery systems of salinomycin / 药学实践杂志

Salinomycin ,extensively used as an antibiotic in animal husbandry for a long time ,has recently been found to possess strong anti-cancer and anti-cancer stem cell efficacy ,as well as activities to overcome multi-drug resistance of tumor based on studies in vivo and in vitro in case reports in pilot clinical trials .Therefore ,salinomycin promised to be a novel anti-cancer agent .However ,the unfavorable property of poor aqueous solubility and the adverse effects of salinomycin were greatly hinder its clinical use .In order to improve its therapeutic index and alleviate its toxicity ,studies on nanotechnology-based deliv-ery systems of salinomycin had been widely conducted .In this article ,the latest development and application of salinomycin nanoformulations were reviewed .

Studies on the preparation of docetaxel nano-liposomes and its treatment on liver cancer cells in vivo and in vitro / 中国生化药物杂志

Objective To prepared the docetaxel nano liposome (L-DOC) for the therapy of liver cancer HepG2 cells in vitro and in vivo. Methods The film-ultrasonic dispersion method was used to prepare the L-DOC.The diameter and Zeta potential of L-DOC were determined by Nanosizer and the encapsulation efficiency was further measured.CCK-8 method was used to determine the cell viability of HepG2 cell after treating with various concentration of DOC and L-DOC respectively and the cell death type was detected by Flow cytometer.Next, we have studied the relative tumor volume change of tumor-bearing mice and the toxicity in vivo.Results The average diameter of L-DOC was 104 nm and the Zeta potential was about -35.1 mV.The Zeta potential of L-DOC was almost unchanged after standing for 96 hours.The encapsulation efficiency of L-DOC was ( 71.2 ±1.6 )%.The CCK-8 results showed that the cell viability was decreased after treating with various concentration of DOC and L-DOC, but the inhibition effect of L-DOC was better than that of DOC after treating with the same dose, especially for 20μg/mL.It was found that the cell death was induced by apoptosis.The in vivo study results showed that 6mg/kg L-DOC could inhibit the tumor volume better than that of same dose of DOC.In addition, 6mg/kg L-DOC and DOC didn’ t induce in vivo toxicity.Conclusion The L-DOC is prepared by film-ultrasonic dispersion method which has small diameter, great biocompatibility.And it could inhibit the HepG2 cells in vitro and in vivo, especially for no in vivo toxicity.

Preparation of palmatine-loaded flexible nano-liposomes and their vaginal mucosa permeability in vitro / 中国药学杂志

OBJECTIVE: To prepare palmatine-loaded flexible nano-liposomes (PFNL) and study their pharmaceutical properties, in order to lay the foundation for the industral application. METHODS: The flexible nano-liposomes were prepared by thin-film homogenization method with propyleneglycol (PG) as softening agent. The entrapment rate of palmatine was evaluated by protamine aggregation method and HPLC. The effects of concentrations of phosphatidylcholine (PSC), cholesterol (CH), and PG on the entrapment efficiency of palmatine were also investigated. The pharmaceutical properties of PFNL were evaluated by TEM, PCS and CLSM. The deformation of PFNL was determined by its relative rate of permeating the microporous filter membrane. The coagulation rate constant (K) was measured by constant temperature conductivity method. The side-by-side diffusion cells and pig vaginal mucosa were used to investigate the characteristics of the release of palmatine from PFNL in vitro, and the effects of PFNL on the expression of cytokines (SLPI, LF and SP-D) were investigated and compared with classic liposomes and lotion of palmatine. RESULTS: The palmatine entrapment efficiency was (78 ±2.13)% when the PFNL were prepared with PSC (3%), CH (0.02%), and PG (20%). The prepared nano flexible liposomes had a closed spherical or elliptical shape and appeared as multi-lamellar vesicles under the TEM and CLSM. The calculated mean size was (185 ±19) nm, and the Zeta potential was (-53 ±2.27) mV. The deformation of PFNL was (79 ± 5.75)%. The coagulation rate constant (K) of PFNL was always lower than that of traditional palmatine- loaded liposomes. The accumulated permeation amount of palmatine from the PFNL at 6.0 h was 1.53 and 2.86 folds of those of classic liposomes and lotion, respectively. Moreover, the expressions of cytokines (SLPI, LF and SP-D) in female SD rats after being treated with PFNL, PCL and PL were similar to that of the control group. CONCLUSION: The prepared PFNL have high encapsulation efficiency, good stability and safety, and greatly increase the vaginal mucosa permeability of palmatine. Flexible nano-liposomes may be a useful drug delivery carrier for the gynecological application of palmatine.

Nanoliposomes of L-lysine-conjugated poly(aspartic acid) Increase the Generation and Function of Bone Marrow-derived Dendritic Cells

BACKGROUND: Biodegradable polymers have increasingly been recognized for various biological applications in recent years. Here we examined the immunostimulatory activities of the novel poly(aspartic acid) conjugated with L-lysine (PLA). METHODS: PLA was synthesized by conjugating L-lysine to aspartic acid polymer. PLA-nanoliposomes (PLA-NLs) were prepared from PLA using a microfluidizer. The immunostimulatory activities of PLA-NLs were examined in mouse bone marrow-derived dendritic cells (BM-DCs). RESULTS: PLA-NLs increased the number of BM-DCs when added to cultures of GM-CSF-induced DC generation on day 4 after the initiation of cultures. Examination of the phenotypic properties showed that BM-DCs generated in the presence of PLA-NLs are more mature in terms of the expression of MHC class II molecules and major co-stimulatory molecules than BM-DCs generated in the absence of PLA-NLs. In addition, the BM-DCs exhibited enhanced capability to produce cytokines, such as IL-6, IL-12, TNF-alpha and IL-1beta. Allogeneic mixed lymphocyte reactions also confirmed that the BMDCs were more stimulatory on allogeneic T cells. PLA- NL also induced further growth of immature BM-DCs that were harvested on day 8. CONCLUSION: These results show that PLA-NLs induce the generation and functional activities of BM-DCs, and suggest that PLA-NLs could be immunostimulating agents that target DCs.

Nanoliposomes of L-lysine-conjugated poly(aspartic acid) Increase the Generation and Function of Bone Marrow-derived Dendritic Cells

BACKGROUND: Biodegradable polymers have increasingly been recognized for various biological applications in recent years. Here we examined the immunostimulatory activities of the novel poly(aspartic acid) conjugated with L-lysine (PLA). METHODS: PLA was synthesized by conjugating L-lysine to aspartic acid polymer. PLA-nanoliposomes (PLA-NLs) were prepared from PLA using a microfluidizer. The immunostimulatory activities of PLA-NLs were examined in mouse bone marrow-derived dendritic cells (BM-DCs). RESULTS: PLA-NLs increased the number of BM-DCs when added to cultures of GM-CSF-induced DC generation on day 4 after the initiation of cultures. Examination of the phenotypic properties showed that BM-DCs generated in the presence of PLA-NLs are more mature in terms of the expression of MHC class II molecules and major co-stimulatory molecules than BM-DCs generated in the absence of PLA-NLs. In addition, the BM-DCs exhibited enhanced capability to produce cytokines, such as IL-6, IL-12, TNF-alpha and IL-1beta. Allogeneic mixed lymphocyte reactions also confirmed that the BMDCs were more stimulatory on allogeneic T cells. PLA- NL also induced further growth of immature BM-DCs that were harvested on day 8. CONCLUSION: These results show that PLA-NLs induce the generation and functional activities of BM-DCs, and suggest that PLA-NLs could be immunostimulating agents that target DCs.