As negative control, white liposomes (WLs) were generated in parallel. (Sigma-Aldrich, catalog number . Then 2.5 g clodronate (Sigma) in 10 mL PBS was added to the lipids. with 200mg/mouse of clodronate liposomes and its control respectively. Clodronate liposomes (FormuMax, Silicon Valley, CA) were injected intraperitoneally one day before initiating the model and the third day after initiating the model to deplete macrophages; control liposomes were used as the control group [27,28,29]. Macrophage Depletion Kit (Standard) Each standard macrophage depletion kit contains one vial of Clodronate encapsulated liposomes (Clodrosome) and one vial of plain liposome for control (Encapsome) with the same lipid composition as Clodrosome. Clodronate liposomes were prepared according to the method by Van Rooijen and Sanders. Read more. PDF IL-15 enhanced antibody-dependent cellular cytotoxicity ... Clodronate Liposomes Improve Metabolic Profile and Reduce ... al. PDF In-anti-F4/80-A3-1 antibody: a novel tracer to image ... Furthermore, an optimization of the drug entrapment into liposomes was performed using polynomial mathematical equations and response surface plots [15-17]. The liposomes contained ap-proximately 2 mg clodronate/ml and 70 μmol total lipid/ml. Conjugates of NK cells with EL4-hCD20 cells NK cells were freshly isolated from WT mice treated with PBS or hIL-15, or with hIL-15 Empty liposomes were used as controls. Clodronate Liposomes. The implication is that Clodronate, once delivered into phagocytic cells using liposomes as vehicles, will not escape . PDF Disease of Aquatic Organisms 57:67 the concept that BM-M8s internalize these liposomes through a phagocytic mechanism resulting in the cross-presentation of the encapsulated OVA by the vacuolar pathway. Dichloromethylene diphosphonate (clodronate, 2.5g; Sigma) was encapsulated in liposomes formed by a 25:1w/w ratio of phos-phatidylcholine:cholesterol (Sigma) as described (resulting liposomes resuspended in 4ml sterile PBS. Mice were fed 20% D-mannose (Sigma-Aldrich, St. Louis, MO) for 7 days [25, 26]. twice a week . clodronate liposomes (i.p. After 12 wk, mice (VEH and CLD) were euthanized, colon contents were collected for microbiota profiling, colons were dissected, and polyps were counted using a stereoscope. Mark Davis,2 James A. Carson,2,4 Cory M. Robinson,3 Liposomes. Furthermore, as expected, clodronate was internalized by J774 macrophages more efficiently when encapsulated in liposomes, since the size of the ultraviolet absorbance peak owing to the metabolite was similar in extracts from cells treated for 24 h with 15 μM liposome-encapsulated clodronate and in extracts from cells treated for 48 h with 250 . 24h later the mice were infection via orbital vein with GBS and 3h after infection, blood was collected for analysis of monocyte number by flow cytometry. While the primary message from the Buiting, et. Clodronate, a member of the bisphosphonate family, is best known as an effective clinical therapy for osteometabolic disorders such as osteoporosis, arthritis, and osteo-articular pain [1-4].When liposome-encapsulated clodronate (clodronate liposomes or Clo-Lip) is injected into an organ, it is phagocytosed by mononuclear phagocytes and then digested by lysosomal enzymes. Mice were injected i.v. Clodronate liposomes improve metabolic profile and reduce visceral adipose macrophage content in diet-induced obese mice. (Sigma-Aldrich) supplemented with 1 μM insulin, 2 mM l-glutamine, 10 IU/mL . Clodronate liposomes were prepared as previously described.22 Briefly, 86 mg phosphatidylcholine (Lipoid EPC; LIPOID, Ludwigshafen, Germany) and 8 mg cholesterol (Sigma Chemical, St Louis, MO) were combined with 10 mL clodronate (0.7 M) solution and sonicated gently. Egg phosphatidylcholine (egg PC) and bovine brain phosphatidylserine (PS) were obtained from Avanti Polar Lipids (Alabaster, AL). Adipose tissue was harvested six days post the first injection for histological studies by performing immunohistochemistry with an anti-F4/80 antibody and for real . PLoS One, 6(9): e24358, 1-11, 2011 . (Sigma-Aldrich), and 150 M of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC; Avanti) by reverse-phase evaporation technique. Note that with 100 Mof lipid/kg clodronate-liposomes, WLs, or PBS. Adipose tissue was harvested six days post the first injection for histological studies by performing immunohistochemistry with an anti-F4/80 antibody and for real . %) and liposomes size (diameter) and identify the main formulation in order to produce stable clodronate liposomes with an appropriate bioavailability after oral administration. When administered intravenously, clodronate-containing liposomes are phagocytosed by the mononuclear phagocyte system, and free clodronate is released from the liposomes to the intracellular region, resulting in destruction of the phagocytic cells. DT was purchased from Sigma-Aldrich. PBS or a 0.6 M solution of clodronate (Sigma) in PBS for 2 h. Resulting multilamellar vesicles were sonicated for 3 min and allowed to swell for 2 h at 25 C. Liposomes were washed 3 times with PBS by centrifugation at 100,000g for 30 min and resus-pended in 4 mL of PBS. Science Park 408 Matrix Innovation Center 1098 XH Amsterdam The Netherlands Phone: +31 (0)20 237 3600 PRODUCT INFORMATION. Dichloromethylenediphosphonic acid disodium salt; CAS Number: 22560-50-5; EC Number: 245-078-9; Synonyms: Clodronic acid disodium salt,Cl2MDP,DMDP,Clodronate disodium; Linear Formula: CH2O6Cl2Na2P2; find Sigma-Aldrich-D4434 MSDS, related peer-reviewed papers, technical documents, similar products & more at Sigma-Aldrich The murine LSL-Kras G12D/+;LSL-Trp53 R172H/+;Pdx-1-Cre (KPC) cell line derived from a pancreatic tumour of a KPC mouse was a kind gift from Dr. A. Neesse, University of Clodronate is not a toxic drug in itself and liposomes (if prepared of phosphatidylcholine and cholesterol) are not toxic either.. Free Clodronate will not easily pass phospholipid bilayers of liposomes and cell membranes but liposomes are eagerly swallowed by macrophages.. Introduction. Briefly, cells were incubated in 96-well plates with liposomes, clodronate and clodrolip (6 h, 37°C, 1 mg clodronate ml −1) and cell viability was determined by addition of WST-1 reagent (Roche . Uptake of liposomal-encapsulated clodronate has been demonstrated by macrophages and monocytes, undergo-ing apoptosis in response to clodronate. Preparation of organotypic hippocampal slice cultures Organotypic hippocampal slice cultures were prepared 25 ng/g on days 1 and 3 before harvest. Encapsula NanoSciences is the formulator and manufacturer of Clodronate liposomes (Clodrosome ® ), control liposomes for Clodrosome (Encapsome ® ), Doxorubicin liposomes (Doxosome ™ ), surface active liposomes for antibody and peptide conjugation (Immunosome ® ), DNA and RNA liposomes (Genesome ® ), fluorescent liposomes (Fluoroliposome . Hence, it may prevent cancer progression by a dual mechanism. Clodronate liposomes (250 µl) were infused i.v. To deplete macrophages, we administered a 200-μl ip injection (∼1 mg) of clodronate-encapsulated liposomes (Clodronate Liposomes, Amsterdam, The Netherlands) beginning at week 6.5 and continuing twice weekly until euthanasia (CLD). Liposomes Prepared in the Presence of p-Aminophenyl Mannose. Because osteoclasts originate from the mononuclear-macrophage system, encapsulation of clodronate into liposomes (clodrolip) resulted in the depletion of in macrophage via apoptosis once they were recognized and . CLD-lips have been used in many studies to deplete monocyte-derived cell populations.11-13 Clodronate is a non-nitrogenous bisphosphonate. Clodronate-loaded liposomal and fibroblast-derived exosomal hybrid system for enhanced drug delivery to pulmonary fibrosis. liposomes treatment, clodronate liposomes combined with DHA treatment. Clodronate is a first-generation bisphosphonate osteoporosis . Kupffer cell depletion was achieved by intravenous injection of 200 lL clodronate liposomes 48 hours before the . [7]. However, liposomal clodronate (LC) has not been previously evaluated in a large-animal spontaneous autoimmune disease model. In brief, 43 mg of L-a-phosphatidylcholine (Sigma-Aldrich) and 4 mg of cholesterol (Nacalai After 48h, when elimination of macrophages in the spleen and Liposomes were extruded 5 times through Cl2MDP (or clodronate) was a gift from Roche (Van Rooijen and Sanders, 1994). Clodronate, a member of the bisphosphonate family, is best known as an effective clinical therapy for osteometabolic disorders such as osteoporosis, arthritis, and osteo-articular pain [1,2,3,4].When liposome-encapsulated clodronate (clodronate liposomes or Clo-Lip) is injected into an organ, it is phagocytosed by mononuclear phagocytes and then digested by lysosomal enzymes. 10 6 wild-type or CD47 − / − RBCs were added, then allowed to settle for 3 min at 4°C, after which sodium pervanadate (Sigma) was added to 2 mM and cells were transferred to a water bath at 37°C. clodronate-liposomes) at 5mg clodronate/ml suspension or liposomes containing PBS (Clodronate Liposomes Foundation, Amsterdam, the Netherlands) were IV injected at .1ml/10g body weight (BW). One method to obtain animal models efficiently depleted in macrophages in different tissues (bone marrow, spleen, liver, lungs, brain, gut, peritoneal cavity, lymph nodes/vessels) and blood is the use of a clodronate-liposome solution.Here, we describe . Treatment of mice with clodronate-liposomes (Clo-lip) or GdCl 3 A suspension of liposomes encapsulating clodronate was prepared according to a method described previously [22, 23]. Macrophages in the liver and spleen can be depleted within 24 h after a single i.v. Briefly, mice were anesthetized by i.p. Cholesterol was purchased from Sigma-Aldrich (USA.). Then the mice were given with 10 mg/kg DHA by oral gavage every other day and 200 µL PBS liposomes or clodronate liposomes (i.p.) This dose was selected based on the recommendations of the manufacturer and previously published literature. To ablate adipose tissue macrophages, clodronate liposomes or PBS liposomes were injected twice into peritoneal cavities of lean or diet-induced obese mice with a three-day interval. drug with potent osteoclast inhibitory activity. To study residual uptake of liposomes, (C) Evaluation of clodronate liposomes on monocyte/macrophage depletion. Dichloromethylene diphosphonate (clodronate, 2.5 g; Sigma) was encapsulated in liposomes formed by a 25:1 w/w ratio of phosphatidylcholine:cholesterol (Sigma) as described , and the resulting liposomes resuspended in 4 ml sterile PBS. While the primary message from the Buiting, et. Read more. Van Rooijen and Sanders described that this final suspension contains about 20 mg of clodronate (0.017 mol/L).29 The tail vein of each mouse was injected with 200 lL of this suspension. vAT was excised after sacrifice and cultured in DMEM (4,5 g/L glucose, L-glutamine and pyruvate; Gibco) supplemented with 10% FBS (GE Healthcare Life Sciences) and 1% penicillin/streptomycin (Gibco) at 37°C on a shaker at 200 rpm. Preparation of liposomes containing clodronate Liposomes containing (dichloromethylene)bisphosphonic acid (clodronate; Sigma-Aldrich, St. Louis, MO, USA) were prepared according to the method of Van Rooijen et al. . (24) C57Bl/6 mice were treated with an intradefect injection (100mL) of control PBS liposomes or osteomac/ macrophage-depleting clodronate liposomes (equal to 7mg/mL of clodronate in suspension) at the time of surgery (n¼9/group), Preparation of Clodronate-Filled Liposomes Liposomes incorporating clodronate were prepared according to the method of Hashimoto et al. They can be used to encapsulate strongly hydropihilic molecules solved in aqueous solutions, such as clodronate, a non-toxic bisphosphonate, developed for human application. Lab technicians: [email protected] SUPPORT. Mice were pre-treated with 200 µL clodronate liposomes (i.p.) The clodronate-containing liposomes will be recognized as foreign particles and get engulfed by macrophages upon dosing into the animal by the chosen routes. Kurume, Japan). Macrophage Depletion Kit (Standard) Each standard macrophage depletion kit contains one vial of Clodronate encapsulated liposomes (Clodrosome) and one vial of plain liposome for control (Encapsome) with the same lipid composition as Clodrosome. At a certain intracellular clodronate concentration, the macrophage is eliminated by apoptosis. Macrophages form a major part of the stromal cell infiltrate in solid tumors and are highly plastic to their environment, which creates phenotypic and functional diversity within the population (1, 4-10).Tumors exploit the plastic nature of macrophages, which we collectively refer to as tumor-associated macrophages (TAMs), to facilitate disease progression by promoting angiogenesis (11, 12 . [26]). Liver resident macrophages designated Kupffer cells (KCs) form the largest subpopulation of tissue macrophages. Phosphate-buffered saline (PBS)-encapsulated liposomes (Clodronate Liposomes) were administered in the same manner to the vehicle control group (VEH). Neutrophil depletion was performed by two intravenous injections of anti-GR1 antibody (100 lg/ dose, RB6-8C5 clone; eBioscience), 24 and 4 hours before the experiment. Clodronate (dichloromethylene bisphosphate, Cl 2MBP; Sigma-Aldrich, St. Louis, MO) was encapsulated in multi-lamellar liposomes, as described.27,35 Clodronate was pre-pared at a concentration of 2.5g per 10mL of PBS solution.35 In the present study, clodronate liposomes were injected IV in the lateral tail vein to deplete macrophages from the pe- Macrophage depletion using clodronate liposomes decreases tumorigenesis and alters gut microbiota in the AOM/DSS mouse model of colon cancer Jackie E. Bader,1 Reilly T. Enos,1 Kandy T. Velázquez,1,2 Meredith S. Carson,1 Mitzi Nagarkatti,1 Prakash S. Nagarkatti,1 Ioulia Chatzistamou,1 J. injection of 0.5 ml clodronate-liposomes, PBS-liposomes or PBS 4 and 2 d prior to analysis, i.e. Clodronate (dichloromethylene bisphosphate, Cl 2MBP; Sigma-Aldrich, St. Louis, MO) was encapsulated in multi-lamellar liposomes, as described.27,35 Clodronate was pre-pared at a concentration of 2.5g per 10mL of PBS solution.35 In the present study, clodronate liposomes were injected IV in the lateral tail vein to deplete macrophages from the . clodronate-liposomes, PBS-liposomes or PBS on Days -4 and -2. Therefore, research-ers have sought to overcome this problem by encasing clodro-nate in liposomes; indeed, clodronate-containing liposomes se- We would like to show you a description here but the site won't allow us. Half of all remaining interstitial ED1 . In addition, SPIO-nanoparticles have been used as cell labeling agents. clodronate liposomes and control liposomes were given on days -1, 1, 4, 7, 11 and 15. In this study, we examined whether KCs are involved in not only intrahepatic inflammation but also extrahepatic systemic inflammation. Once anesthetized, mice were injected with 100 L clodronate liposomes intrarectally using a micropipette. cholesterol (Sigma-Aldrich, St Louis, MO, USA) as described previously. Liposome-encapsulated clodronate Liposomes containing clodronate (i.e. A major constraint in the study of Plasmodium falciparum malaria, including vaccine development, lies on the parasite's strict human host specificity and therefore the shortage of animal experimental models able to harbor human plasmodia. Briefly, 75 mg phosphatidylcholine However, the role of KCs in the systemic inflammation is still elusive. Prolonged Hepatomegaly in Mice That Cannot Inactivate Bacterial Endotoxin Baomei Shao,1 Richard L. Kitchens,1 Robert S. Munford,1,2 Thomas E. Rogers,3 Don C. Rockey,4 and Alan W. Varley1 Transient hepatomegaly often accompanies acute bacterial infections. Clodronate molecules are encapsulated in small liposomes, which have excellent activity, physical and chemical stability, and are easy to use and handle. . Cholesterol was purchased from Sigma (St. Louis, MO) and clodronate was a . Only 1-2% of clodronate becomes encapsulated, yielding an estimated dose of 0.7-1.0 mg clodronate per 100 . U/mouse, Sigma) were given 6 and 18 hours after APAP challenge. Only 1-2% of clodronate becomes encapsulated, yielding an estimated dose of 0.7-1.0 mg clodronate per 100 . Liposomes Clodronate (Sifavitor) and rhodamine RE (Avanti Polar Lipids) were encapsulated in liposomes composed of 50 mol/L distearoyl-phosphatidylglycerol (DSPG) (Avanti), 100 mol/L cholesterol (Sigma Chemicals), and 150 mol/L of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) (Avanti) by reverse-phase evaporation tech- Grafts were harvested on day 5. Liposomes were diluted in 300 µl phosphate-buffered saline (PBS) injection of 200 L 2.5% Avertin per mouse. paper is the the p-aminophenyl mannose formulation or other reformulations did not demonstrate a significantly different biodistribution or behavior compared to unmodified clodronate liposome formulations, we do not believe that the authors addressed the issue of clodronate liposome . Clodronate Liposomes. Clodronate (CLD) was the first generation of bisphosphonates, used for the treatment of osteoporosis and osteolytic metastasis [10]. Clodronate was a gift of Roche Diagnostics (Man-nheim, Germany), phosphatidylcholine (Lipoid EPC) was obtained from Lipoid (Ludwigshafen, Germany) and cho-lesterol was purchased from Sigma (USA). al. After a single intravenous or intraperitoneal administration of clodronate liposomes, 90% of macrophages in the spleen can be depleted. Clodronate (Sifavitor) and rhodamine RE (Avanti Polar Lipids) were encapsulated in liposomes composed of 50 μmol/L distearoyl-phosphatidylglycerol (DSPG) (Avanti), 100 μmol/L cholesterol (Sigma Chemicals), and 150 μmol/L of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) (Avanti) by reverse-phase evaporation technique, as described elsewhere. After 12 wk, mice (VEH and CLD) were euthanized, colon contents were collected for microbiota profiling, colons were dissected, and polyps were counted using a stereoscope. However, clodro-nate has poor cell membrane permeability. Clodronate-liposomes deplete CD169-expressing macrophages within the BM. s2A. 9 The average size of liposomes was . k Agouti renal allograft and liposomal-clodronate, control phosphate-buffered saline liposomes, or saline intravenously (n=7 per group) on days 1 and 3 postsurgery. Liposomal clodronate (dichloromethylene diphosphonate) has been used to deplete macrophages and block clearance of opsonized cells in mouse models of autoimmune disease. The liposomes were resuspended in 4 mL PBS. Freely solved clodronate will not cross . Doxil®, a formulation of DXR encapsulated in polyethylene glycol modified (PEGylated) liposomes, is the most widely used of the clinically approved liposomal anticancer drugs. The resulting liposomes were then washed to eliminate free drug. Thus, CLD-lip treatment preferentially Clodronate molecules are encapsulated in small liposomes, which have excellent activity, physical and chemical stability, and are easy to use and handle. Empty liposomes were prepared in the same manner using PBS instead of the aqueous clodronate solution. Treatment of wild-type mice with clodronate liposomes . 50 ng cholesterol and 8 mg egg lecithin (L-α-Phosphatidyl-DL-glycerol sodium salt (Sigma P8318)) were dissolved in chloroform in a round-bottom flask. Marketing & Administration support: Sigma Chemical Co. (St. Louis, MO). Article Snippet: To determine the short-term effect of bisphosphonates on ECM synthesis, cultures were supplemented with media containing either clodronate or pamidronate (Sigma-Aldrich Ltd.) 2 days after seeding at varying concentrations (0, 1, 10, and 100 μM) and incubated in the presence of both [35 S]SO4 (5 μCi/culture) to label . Lifespan, Rhode Island's first health system, was founded in 1994 by Rhode Island Hospital and the Miriam Hospital. at 3 days before the induction of colitis. Clodronate Liposomes Improve Metabolic Profile and Reduce Visceral Adipose Macrophage Content in Diet-Induced Obese Mice Bin Feng1,3, Ping Jiao3, Yaohui Nie3, Thomas Kim3, Dale Jun3, Nico van Rooijen2, Zaiqing Yang1, Haiyan Xu3* 1Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Life Science and Technology, Huazhong Agricultural Preparation of clodronate liposomes: The preparation of clodronate liposomes was similar to previous descriptions (Van Rooijen 1989; Zelinka et al. Total BM was isolated from long bones after 24 hours (A, B) or as indicated (C); stained with Cd11b, F4/80, and CD169 fluorescently conjugated antibodies; and analyzed by flow cytometry. Appendix 1 Supplemental Experimental Procedures: Cell culture All cells were cultured in a humidified atmosphere containing 5% CO 2 at 37°C. at 1 d (14 h), 7 d, 10 d, 16 d, or 28 d before harvest. Phosphate-buffered saline (PBS)-encapsulated liposomes (Clodronate Liposomes) were administered in the same manner to the vehicle control group (VEH). ADRESS INFORMATION. In order to decipher the role of macrophages in vivo, it could be necessary to establish a model of macrophage depletion in the whole animal.

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