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Worm-on-a-chip: Fully automated whole organism platform for screening and identification of toxicity using the nematode Caenorhabditis elegans
P-09-01 / Toxicology Letters 280S (2017) S249–S253
P-09-01-06 Optimization of semi-occlusive dermal exposure method in pregnant and lactating rats Kirsten Hartman Van Dycke, Francois van Otterdijk, Chantal Beerens, Harrie van der Eerden, Harry Emmen, Manon Beekhuijzen Charles River Den Bosch, ’s Hertogenbosch, Netherlands A commonly used method for (semi) occlusive dermal exposure in rats is full body wrap, for which the correct tightness is essential. The consequences of imbalance range from potential mortality to detachment of the wrapping and consequent insufficient dermal exposure and increased oral exposure. Additional practical and scientific challenges arise when implemented in pregnant and lactating rats. Our aim was to overcome these technical difficulties and improve animal welfare, by setting up a method for semiocclusive dermal exposure for use during gestation and lactation. A dermal patch consisting of a gauze dressing on non-irritating tape covered by a film dressing was applied on the dorsal surface of the animals. These dermal patch applications were combined with Lomir jackets including dermal inserts and tested in 2 male and 2 female Wistar Han rats. Initially, the animals were trained to wear the Lomir jackets for up to two weeks and subsequently animals were treated for 6 h per day with 1 mL of vehicle (corn oil) during pre-mating, mating, gestation up to Day 19 post-coitum and lactation from Day 4 onwards. During exposure pups were kept with their dams. Development of body weight for the parental animals and pups were normal throughout the experiment, and skin irritation was limited and transient. No difficulties were observed during lactation. In conclusion, the described method proves successful for dermal exposure in pregnant and lactating rats, and is recommended as alternative for the full body wrap method because of improved animal welfare. http://dx.doi.org/10.1016/j.toxlet.2017.07.694 P-09-01-07 Acute and subchronic oral toxicity study of antihypertensive polyherbal preparation, Herbamon Eti Nurwening Sholikhah, Mustofa Mustofa, Dwi Aris Agung Nugrahaningsih, Fara Silvia Yuliani, Setyo Purwono, Ngatidjan Ngatidjan Department of Pharmacology and Therapy, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia Herbamon is Indonesian polyherbal preparation as antihypertensive agent which containing: Allium sativum, Belericae fructus, Curcumae aeruginosae, and Amomi fructus. This study was conducted to evaluate the acute and subchronic oral toxicity of Herbamon in rats. The acute toxicity study was conducted on 6 female Wistar rats using fixed dose method. The preliminary study on one rat with dose of 300 mg/kg did not show any sign and symptoms of toxicity, so that the preliminary study continued on one rat with dose of 2000 mg/kg. The preliminary study with dose of 2000 mg/kg did not show any toxicity signs and symptoms as well. So that, the study continued at dose of 2000 mg/kg on other 4 rats. Each animal was observed for the first 24 h and continued for 14 days. There were no significant toxic effects and no death observed until the end of the study, showed that the lethal dose 50% (LD50) of Herbamon was >2000 mg/kg. The macroscopic and microscopic examination of internal organs showed no symptoms of toxicity.
S251
At the subchronic toxicity study, the Herbamon with doses of 126, 252, and 1000 mg/kg per day were administered on 80 Wistar rats for 90 days orally. There were no significant toxic effects observed at all dose on symptom, macroscopic and microscopic examination. These findings showed that the long term oral administration of polyherbal preparation, Herbamon for 90 days did not cause subchronic toxicity. http://dx.doi.org/10.1016/j.toxlet.2017.07.695 P-09-01-08 Worm-on-a-chip: Fully automated whole organism platform for screening and identification of toxicity using the nematode Caenorhabditis elegans Robert Mader 1 , Laurent Mouchiroud 2 , Matteo Cornaglia 3 , Martin A.M. Gijs 3 , Johan Auwerx 2 1 Translational Medicine, Debiopharm International SA, Lausanne, Switzerland 2 Laboratory of Integrative and Systems Physiology, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland 3 Laboratory of Microsystems, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland
Currently, a gap exists between in vitro assays capable of detecting specific types of toxicity and animal toxicity studies allowing identification of several different toxicities at once. From ethical and financial aspects, the frequent use of vertebrates to identify toxicities in a screening program is questionable. In this context, the roundworm Caenorhabditis elegans appears as an attractive alternative, offering a good compromise between the simplicity of cellular models and the complexity of vertebrates. It has previously demonstrated predictive value for mammalian toxicity. The protocols used to date rely almost entirely on laborious manual handling and time-consuming direct observation by the operator. Therefore, we have developed an innovative fully automated platform for worm handling and observation, combined with dedicated software for data collection and analysis. Our microfluidic device has been developed with the goal of automated medium throughput high-content phenotyping of worms by recording several parameters in real-time. This model requires only minute amounts of compound and can potentially identify the mechanism of toxicity through specific phenotypic patterns within 3 days. To demonstrate the utility, we validated our platform by screening compounds with well-defined toxicity profiles. By recording several parameters (e.g. larval growth, fertility, mobility) over 3 days, we were able to identify distinct patterns related to specific toxicity profiles of the tested compounds. In conclusion, we propose an innovative solution for rapid identification of toxic compounds and their potential mechanism of toxicity in a model bridging the gap between in vitro and in vivo assays. http://dx.doi.org/10.1016/j.toxlet.2017.07.696
P-09-01-06 Optimization of semi-occlusive dermal exposure method in pregnant and lactating rats Kirsten Hartman Van Dycke, Francois van Otterdijk, Chantal Beerens, Harrie van der Eerden, Harry Emmen, Manon Beekhuijzen Charles River Den Bosch, ’s Hertogenbosch, Netherlands A commonly used method for (semi) occlusive dermal exposure in rats is full body wrap, for which the correct tightness is essential. The consequences of imbalance range from potential mortality to detachment of the wrapping and consequent insufficient dermal exposure and increased oral exposure. Additional practical and scientific challenges arise when implemented in pregnant and lactating rats. Our aim was to overcome these technical difficulties and improve animal welfare, by setting up a method for semiocclusive dermal exposure for use during gestation and lactation. A dermal patch consisting of a gauze dressing on non-irritating tape covered by a film dressing was applied on the dorsal surface of the animals. These dermal patch applications were combined with Lomir jackets including dermal inserts and tested in 2 male and 2 female Wistar Han rats. Initially, the animals were trained to wear the Lomir jackets for up to two weeks and subsequently animals were treated for 6 h per day with 1 mL of vehicle (corn oil) during pre-mating, mating, gestation up to Day 19 post-coitum and lactation from Day 4 onwards. During exposure pups were kept with their dams. Development of body weight for the parental animals and pups were normal throughout the experiment, and skin irritation was limited and transient. No difficulties were observed during lactation. In conclusion, the described method proves successful for dermal exposure in pregnant and lactating rats, and is recommended as alternative for the full body wrap method because of improved animal welfare. http://dx.doi.org/10.1016/j.toxlet.2017.07.694 P-09-01-07 Acute and subchronic oral toxicity study of antihypertensive polyherbal preparation, Herbamon Eti Nurwening Sholikhah, Mustofa Mustofa, Dwi Aris Agung Nugrahaningsih, Fara Silvia Yuliani, Setyo Purwono, Ngatidjan Ngatidjan Department of Pharmacology and Therapy, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia Herbamon is Indonesian polyherbal preparation as antihypertensive agent which containing: Allium sativum, Belericae fructus, Curcumae aeruginosae, and Amomi fructus. This study was conducted to evaluate the acute and subchronic oral toxicity of Herbamon in rats. The acute toxicity study was conducted on 6 female Wistar rats using fixed dose method. The preliminary study on one rat with dose of 300 mg/kg did not show any sign and symptoms of toxicity, so that the preliminary study continued on one rat with dose of 2000 mg/kg. The preliminary study with dose of 2000 mg/kg did not show any toxicity signs and symptoms as well. So that, the study continued at dose of 2000 mg/kg on other 4 rats. Each animal was observed for the first 24 h and continued for 14 days. There were no significant toxic effects and no death observed until the end of the study, showed that the lethal dose 50% (LD50) of Herbamon was >2000 mg/kg. The macroscopic and microscopic examination of internal organs showed no symptoms of toxicity.
S251
At the subchronic toxicity study, the Herbamon with doses of 126, 252, and 1000 mg/kg per day were administered on 80 Wistar rats for 90 days orally. There were no significant toxic effects observed at all dose on symptom, macroscopic and microscopic examination. These findings showed that the long term oral administration of polyherbal preparation, Herbamon for 90 days did not cause subchronic toxicity. http://dx.doi.org/10.1016/j.toxlet.2017.07.695 P-09-01-08 Worm-on-a-chip: Fully automated whole organism platform for screening and identification of toxicity using the nematode Caenorhabditis elegans Robert Mader 1 , Laurent Mouchiroud 2 , Matteo Cornaglia 3 , Martin A.M. Gijs 3 , Johan Auwerx 2 1 Translational Medicine, Debiopharm International SA, Lausanne, Switzerland 2 Laboratory of Integrative and Systems Physiology, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland 3 Laboratory of Microsystems, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland
Currently, a gap exists between in vitro assays capable of detecting specific types of toxicity and animal toxicity studies allowing identification of several different toxicities at once. From ethical and financial aspects, the frequent use of vertebrates to identify toxicities in a screening program is questionable. In this context, the roundworm Caenorhabditis elegans appears as an attractive alternative, offering a good compromise between the simplicity of cellular models and the complexity of vertebrates. It has previously demonstrated predictive value for mammalian toxicity. The protocols used to date rely almost entirely on laborious manual handling and time-consuming direct observation by the operator. Therefore, we have developed an innovative fully automated platform for worm handling and observation, combined with dedicated software for data collection and analysis. Our microfluidic device has been developed with the goal of automated medium throughput high-content phenotyping of worms by recording several parameters in real-time. This model requires only minute amounts of compound and can potentially identify the mechanism of toxicity through specific phenotypic patterns within 3 days. To demonstrate the utility, we validated our platform by screening compounds with well-defined toxicity profiles. By recording several parameters (e.g. larval growth, fertility, mobility) over 3 days, we were able to identify distinct patterns related to specific toxicity profiles of the tested compounds. In conclusion, we propose an innovative solution for rapid identification of toxic compounds and their potential mechanism of toxicity in a model bridging the gap between in vitro and in vivo assays. http://dx.doi.org/10.1016/j.toxlet.2017.07.696