Bi-Functional Radiotheranostics of 188Re-Liposome-Fcy-hEGF for Radio- and Chemo-Therapy of EGFR-Overexpressing Cancer Cells

Epidermal growth factor receptor (EGFR) specific therapeutics is of great importance in cancer treatment. Fcy-hEGF fusion protein, composed of yeast cytosine deaminase (Fcy) and human EGF (hEGF), is capable of binding to EGFR and enzymatically convert 5-fluorocytosine (5-FC) to 1000-fold toxic 5-fluorocuracil (5-FU), thereby inhibiting the growth of EGFR-expressing tumor cells. To develop EGFR-specific therapy, ¹⁸⁸Re-liposome-Fcy-hEGF was constructed by insertion of Fcy-hEGF fusion protein onto the surface of liposomes encapsulating of ¹⁸⁸Re. Western blotting, MALDI-TOF, column size exclusion and flow cytometry were used to confirm the conjugation and bio-activity of ¹⁸⁸Re-liposome-Fcy-hEGF. Cell lines with EGFR expression were subjected to treat with ¹⁸⁸Re-liposome-Fcy-hEGF/5-FC in the presence of 5-FC. The ¹⁸⁸Re-liposome-Fcy-hEGF/5-FC revealed a better cytotoxic effect for cancer cells than the treatment of liposome-Fcy-hEGF/5-FC or ¹⁸⁸Re-liposome-Fcy-hEGF alone. The therapeutics has radio- and chemo-toxicity simultaneously and specifically target to EGFR-expression tumor cells, thereby achieving synergistic anticancer activity.     

Intravitreal Injection of Liposomes Loaded with a Histone Deacetylase Inhibitor Promotes Retinal Ganglion Cell Survival in a Mouse Model of Optic Nerve Crush

Various neuroprotective agents have been studied for the treatment of retinal ganglion cell (RGC) diseases, but issues concerning the side effects of systemically administered drugs and the short retention time of intravitreally injected drugs limit their clinical applications. The current study aimed to evaluate the neuroprotective effects of intravitreally injected trichostatin A (TSA)-loaded liposomes in a mouse model of optic nerve crush (ONC) and determine whether TSA-loaded liposomes have therapeutic potential in RGC diseases. The histone deacetylase inhibitor, TSA, was incorporated into polyethylene glycolylated liposomes. C57BL/6J mice were treated with an intravitreal injection of TSA-loaded liposomes and liposomes loaded with a lipophilic fluorescent dye for tracking, immediately after ONC injury. The expression of macroglial and microglial cell markers (glial fibrillary acidic protein and ionized calcium binding adaptor molecule-1), RGC survival, and apoptosis were assessed. We found that the liposomes reached the inner retina. Their fluorescence was detected for up to 10 days after the intravitreal injection, with peak intensity at 3 days postinjection. Intravitreally administered TSA-loaded liposomes significantly decreased reactive gliosis and RGC apoptosis and increased RGC survival in a mouse model of ONC. Our results suggest that TSA-loaded liposomes may help in the treatment of various RGC diseases.     

Influence of Sodium Alginate and Genipin on Stability of Chitosome Containing Perilla Oil in Model and Real Drink

The impacts of sodium alginate (SA) and genipin (GP) on low‐pH and thermal stability of chitosan‐coated liposome (chitosome), as a carrier of perilla oil, are investigated in a model drink. Oxidative stability, as well as quality parameters and sensory evaluation, are analyzed during 60 days of storage at 25 °C as well as in orange drink fortified with non‐coated and coated nanoliposome. The investigation of model drink shows that low‐pH and heating stability of nanoliposome are influenced by the type of coating biopolymer and can be improved by chitosan (CS) and GP. The coating by different biopolymers has fewer changes in quality parameters (°Brix and pH value, total acidity, and color difference), and shows higher oxidative stability than emulsified perilla seed oil and bare nanoliposome after storage at 25 °C for 60 days. The results indicate that the type of biopolymer and cross‐linkers plays a key role in the liposomal membrane structure stability. Practical applications: The present study is anticipated to promote a better understanding of the advantages of combining nanoliposomes (NLs) with the primary and secondary coatings as well as cross‐linkers to overcome the low oxidative stability of perilla oil (as a source of plant rich in n‐3 polyunsaturated fatty acids [PUFAs]) and semi‐permeability of NL membrane in like and real foods. It could also reduce the adverse effects of n‐3 PUFAs on sensory properties of orange drink and pave the way for making liposomes applicable to foods and drinks.     

A Step-by-Step Approach to Improve Clinical Translation of Liposome-Based Nanomaterials,a Focus on Innate Immune and Inflammatory Responses

This study aims to provide guidelines to design and perform a robust and reliable physical-chemical characterization of liposome-based nanomaterials, and to support method development with a specific focus on their inflammation-inducing potential. Out of eight differently functionalized liposomes selected as “case-studies”, three passed the physical-chemical characterization (in terms of size-distribution, homogeneity and stability) and the screening for bacterial contamination (sterility and apyrogenicity). Although all three were non-cytotoxic when tested in vitro, they showed a different capacity to activate human blood cells. HSPC/CHOL-coated liposomes elicited the production of several inflammation-related cytokines, while DPPC/CHOL- or DSPC/CHOL-functionalized liposomes did not. This work underlines the need for accurate characterization at multiple levels and the use of reliable in vitro methods, in order to obtain a realistic assessment of liposome-induced human inflammatory response, as a fundamental requirement of nanosafety regulations.     

Acid-Sphingomyelinase Triggered Fluorescently Labeled Sphingomyelin Containing Liposomes in Tumor Diagnosis after Radiation-Induced Stress

In liposomal delivery, a big question is how to release the loaded material into the correct place. Here, we will test the targeting and release abilities of our sphingomyelin-consisting liposome. A change in release parameters can be observed when sphingomyelin-containing liposome is treated with sphingomyelinase enzyme. Sphingomyelinase is known to be endogenously released from the different cells in stress situations. We assume the effective enzyme treatment will weaken the liposome making it also leakier. To test the release abilities of the SM-liposome, we developed several fluorescence-based experiments. In in vitro studies, we used molecular quenching to study the sphingomyelinase enzyme-based release from the liposomes. We could show that the enzyme treatment releases loaded fluorescent markers from sphingomyelin-containing liposomes. Moreover, the release correlated with used enzymatic activities. We studied whether the stress-related enzyme expression is increased if the cells are treated with radiation as a stress inducer. It appeared that the radiation caused increased enzymatic activity. We studied our liposomes’ biodistribution in the animal tumor model when the tumor was under radiation stress. Increased targeting of the fluorescent marker loaded to our liposomes could be found on the site of cancer. The liposomal targeting in vivo could be improved by radiation. Based on our studies, we propose sphingomyelin-containing liposomes can be used as a controlled release system sensitive to cell stress.     

Influence of stabilising agents on the properties of liposomal encapsulated ethanolic coconut husk extract

Characteristics of liposomal encapsulated ethanolic coconut husk extract (LE-ECHE) as influenced by different stabilising agents (SA) were investigated. LE-ECHE was prepared using four liposome formulations including soy phosphatidylcholine (SPC) alone and SPC mixed with cholesterol (CHL), tween 80 (TWE) or glycerol (GLY) at a ratio of 4 : 1 as lipid phase (LP) at a concentration of 60 µmol mL⁻¹. Particle size, zeta potential, poly-dispersity index, encapsulation efficiency (EE) and suspension stability (SS) of LE-ECHE samples were markedly influenced by SA (P < 0.05). Antioxidant activity of ECHE was retained after entrapment in liposome, regardless of SA used. However, SA affected the antioxidant activities of LE-ECHE differently. Antibacterial properties of ECHE were enhanced after encapsulation in liposome, irrespective of SA. Therefore, liposome with glycerol or cholesterol could serve as a delivery based system for improving the antibacterial properties with high stability, while lowering the undesirable colour of ECHE.     

Impact of Liposomal Drug Formulations on the RBCs Shape,Transmembrane Potential,and Mechanical Properties

Liposomal technologies are used in order to improve the effectiveness of current therapies or to reduce their negative side effects. However, the liposome–erythrocyte interaction during the intravenous administration of liposomal drug formulations may result in changes within the red blood cells (RBCs). In this study, it was shown that phosphatidylcholine-composed liposomal formulations of Photolon, used as a drug model, significantly influences the transmembrane potential, stiffness, as well as the shape of RBCs. These changes caused decreasing the number of stomatocytes and irregular shapes proportion within the cells exposed to liposomes. Thus, the reduction of anisocytosis was observed. Therefore, some nanodrugs in phosphatidylcholine liposomal formulation may have a beneficial effect on the survival time of erythrocytes.     

应用脂质体紫杉醇联合顺铂对比白蛋白结合型紫杉醇联合顺铂一线治疗晚期非小细胞肺癌的临床观察

目的：观察脂质体紫杉醇联合顺铂对比白蛋白结合型紫杉醇联合顺铂一线治疗晚期非小细胞肺癌（NSCLC）的临床疗效及预后,同时评价血清乳酸脱氢酶（LDH）在预后的评价的作用。方法62例经病理学证实为晚期的NSCLC患者,按随机数字表法分为白蛋白结合型紫杉醇联合顺铂组（A组,31例）和脂质体紫杉醇联合顺铂组（B组,31例）。分析2组的疗效、不良反应及远期生存。测定血清乳酸脱氢酶（LDH）,并根据化疗后血清LDH变化,分为反应组（下降）和不反应组（升高或持平）。结果A组和B组有效率分别为12．9％和25．8％,疾病控制率分别为90．3％和93．5％,2组比较差异无统计学意义（P＞0．05）。A组中位PFS为17．4个月优于B组9．6个月（95％可信区间,5．2个月to19．7个月,P＝0．264）。亚组分析鳞癌患者应用白蛋白结合型紫杉醇联合顺铂方案化疗优于脂质体紫杉醇联合顺铂。2组主要不良反应为骨髓抑制和消化道反应。Cox多因素分析提示性别、年龄、LDH反应性均是影响预后的独立危险因素;女患者预后较男患者好（HR＝3．231,P＝0．014）,初治时年龄大于60岁的患者预后较好（HR＝0．287,P＝0．005）,LDH反应组比不反应组预后好（HR＝0．379,P＝0．027）。结论脂质体紫杉醇联合顺铂和白蛋白结合型紫杉醇联合顺铂均为较好的治疗方案,LDH反应性可能预测晚期NSCLC患者疗效。     

Modulation of Induced Cytotoxicity of Doxorubicin by Using Apoferritin and Liposomal Cages

Doxorubicin is an effective chemotherapeutic drug, however, its toxicity is a significant limitation in therapy. Encapsulation of doxorubicin inside liposomes or ferritin cages decreases cardiotoxicity while maintaining anticancer potency. We synthesized novel apoferritin- and liposome-encapsulated forms of doxorubicin (“Apodox” and “lip-8-dox”) and compared its toxicity with doxorubicin and Myocet on prostate cell lines. Three different prostatic cell lines PNT1A, 22Rv1, and LNCaP were chosen. The toxicity of the modified doxorubicin forms was compared to conventional doxorubicin using the MTT assay, real-time cell impedance-based cell growth method (RTCA), and flow cytometry. The efficiency of doxorubicin entrapment was 56% in apoferritin cages and 42% in the liposome carrier. The accuracy of the RTCA system was verified by flow-cytometric analysis of cell viability. The doxorubicin half maximal inhibition concentrations (IC₅₀) were determined as 170.5, 234.0, and 169.0 nM for PNT1A, 22Rv1, and LNCaP, respectively by RTCA. Lip8-dox is less toxic on the non-tumor cell line PNT1A compared to doxorubicin, while still maintaining the toxicity to tumorous cell lines similar to doxorubicin or epirubicin (IC₅₀ = 2076.7 nM for PNT1A vs. 935.3 and 729.0 nM for 22Rv1 and LNCaP). Apodox IC₅₀ was determined as follows: 603.1, 1344.2, and 931.2 nM for PNT1A, 22Rv1, and LNCaP.     

影响补铁剂脂质体脂质氧化的因素分析

脂质氧化是补铁剂脂质体稳定性的主要指标之一,为明确内外因素对补铁剂脂质体脂质氧化的影响,以TBA法测定脂质体中丙二醛(MDA)含量,并以此为评价指标,考察了壁材组成、芯壁比、芯材类型、p H、温度、超声等主要因素对补铁剂脂质体脂质氧化的影响。实验结果显示,在壁材组成(蛋黄卵磷脂:胆固醇,EPC∶CHOL)为8∶1(wt./wt.)时脂质氧化程度最低;随着芯壁比的降低脂质氧化程度下降;越短、超声功率越低越能有效降低补铁剂脂质体的脂质氧化程度,提高其稳定性。这为补铁剂脂质体的合理生产,以及提高其贮藏和应用过程中的稳定性提供了科学依据。