As soon as tumors reached a volume of approximately 500 mm3, 5 x 105 HSP70B-NIS-MSCs were injected systemically via the tail vein every second day for a total of three times. 3 days later, regional hyperthermia was applied (41 °C or, as control, 37 °C for 1 h). The mice received 18.5 MBq (0.5 mCi) of 123I (GE Healthcare Buchler GmBH & Co. KG, Braunschweig, Germany) intraperitoneally (i.p.) after 0, 6, 12, 18, 24, 36, 48, and 72 h and gamma camera imaging (e.cam, Siemens, Munich, Germany) was performed using a low-energy, high-resolution collimator. Intrinsic thyroidal iodide uptake was reduced by the addition of 5 mg/ml L-thyroxine (L-T4; Sigma Aldrich) to the drinking water ten days before 123I administration. Using the HERMES GOLD (Hermes Medical Solutions, Stockholm, Sweden) software, regions of interest were evaluated and tumoral iodide uptake was calculated and expressed as percentage of injected dose (ID) per tumor (% ID/tumor). Using the Medical Internal Radiation Dose (MIRD) concept, dosimetry was calculated with a RADAR dose factor (www.doseinfo-radar.com ).
L thyroxine l t4
Manufactured by Merck Group
Sourced in United States
L-thyroxine (L-T4) is a synthetic form of the thyroid hormone thyroxine. It is a prescription medication used to treat hypothyroidism, a condition where the thyroid gland does not produce enough thyroid hormones. L-T4 works by replacing the missing thyroid hormone and restoring normal thyroid function.
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Lab products found in correlation
Inveon, by Siemens (2 mentions)
Inveon research workplace, by Siemens (2 mentions)
Inveon acquisition workplace, by Siemens (2 mentions)
Hermes gold, by Hermes Medical Solutions (1 mentions)
Hydrogen peroxide h2o2, by Merck Group (1 mentions)
Thiobarbituric acid, by Merck Group (1 mentions)
Sodium dodecyl sulphate (sds), by Merck Group (1 mentions)
M phosphoric acid, by Merck Group (1 mentions)
6 propyl 2 thiouracil ptu, by Merck Group (1 mentions)
6 protocols using l thyroxine l t4
1
Targeted Hyperthermia-Enhanced Radioiodine Therapy
2
Targeted NIS-Mediated Tumor Imaging
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At 3.5–4 weeks after i.c. tumor cell inoculation, polyplexes (monoDBCO-PEG24-GE11/NIS and bisDBCO-PEG24-GE11/NIS polyplexes for EGFR targeting, non-targeted monoDBCO-PEG24/NIS and bisDBCO-PEG24/NIS polyplexes, monoDBCO-PEG24-GE11/LUC and bisDBCO-PEG24-GE11/LUC containing pCMVLuc as additional negative control) with a DNA dose of 2.5 mg/kg (for a 20-g mouse, 50 μg DNA in a total volume of 250 μL; solvent, HBG) were applied systemically via the tail vein. After 24 or 48 h, mice received 10 MBq of 124I (PerkinElmer, Waltham, MA, or DSD Pharma, Purkersdorf, Austria) as an NIS PET tracer by i.v. injection, and NIS-mediated iodide accumulation in tumor areas was determined by small-animal PET (Inveon, SIEMENS Preclinical Solutions, Erlangen, Germany). Serial scanning took place 1, 3, and 5 h after 124I application. Results were assessed with the software Inveon Acquisition Workplace (Siemens, Munich, Germany), were analyzed using Inveon Research Workplace (Siemens), and are represented as a percentage of the injected dose per milliliter tumor (% ID/mL). Mice were pretreated with L-thyroxine (LT4; 5 mg/mL, Sigma Aldrich) in their drinking water 10 days before imaging to reduce thyroidal iodide uptake, and at the same time the mouse chow was changed to a low-iodine diet (ssniff Spezialdiäten GmbH, Soest, Germany).
3
Establishment and Characterization of SCH Rat Model
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Establishment of the SCH rat model refers to the previously literature [13 (link)].The rats were injected with 3% pentobarbital sodium (0.1 mL/100 g) and underwent thyroidectomy, while Sham group rats underwent sham thyroid surgery. The rats were fed normally for 4 w after operation, then the blood was collected from the retroorbital venous plexus, and serum TSH and TT4 were detected. When serum TSH levels were higher than that in Sham group, the TT4 levels were lower than that in Sham group, confirming the successful establishment of the SCH rat model. Four weeks after surgery, rats in the SCH group were injected subcutaneously with L-thyroxine (L-T4, Sigma, USA) 1.0 μg/100 g/day on the neck. Sham group rats were injected subcutaneously with physiological saline (50 μL/100 g/day) on the neck. Calcium lactate (0.1% w/v) was added to the drinking water for all rats after surgery. Nine days later, all rats were mated with normal male rats (male: female = 1: 2). The pregnant rats were then kept in single cages until delivery. The day of vaginal plus was confirmed by microscopic observation and designated as E0. Serum and tissue samples were collected at E16, E18, P5 and P10. At the end of the experiment, all rats were anesthetized with pentobarbital (50 mg/kg, intraperitoneal) and euthanized by thoracotomy and hearts removal.
4
NIS-Mediated PET Imaging of Tumor Targeting
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Experiments started 5-6 weeks after intrahepatic injection of tumor cells. For determination of tumor specific NIS expression and subsequent NIS-mediated iodide uptake, animals received LPEI-PEG-GE11/NIS polyplexes for EGFR-targeting (n = 6), LPEI-PEG-cMBP/NIS for cMETtargeting (n = 4) or both for dual-targeting (n = 7). Polyplexes were administered systemically via the tail vein (intravenously, i.v.) at a DNA dose of 2.5 mg/kg (50 μg DNA in 250 μL HBG). At 48 h after polyplex injection mice received the NIS-specific PET tracer 124 I and accumulation in tumor tissue was determined by small-animal PET (Inveon, SIEMENS Preclinical Solutions, Erlangen, Germany). Serial scanning was performed after 1, 3, and 5 h. Regions of interest were analyzed with the software Inveon Acquisition Workplace (Siemens), quantified using Inveon Research Workplace (Siemens) and expressed as a fraction of the total amount of initial dose (% of ID). To suppress thyroidal iodide uptake, a 10-day pretreatment with L-thyroxine (L-T4; 5 mg/ml; Sigma-Aldrich) in drinking water was conducted before PET-imaging. To verify NIS-specific uptake, pretreatment with an intraperitoneal (i.p.) injection of 2 mg of the competitive NIS inhibitor sodium perchlorate (NaClO 4 ) 30 min before PET tracer administration (n = 2) was performed.
5
Thyroid Hormone Estimation Protocol
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L-thyroxine (L-T 4 ) was purchased from Sigma-Aldrich chemicals (St. Louis, MO, USA), India. While Elman's reagent, m-phosphoric acid, thiobarbituric acid (TBA), sodium dodecyl sulphate, tricarboxylic acid and hydrogen peroxide (H 2 O 2 ) were obtained from E. Merck Ltd., Mumbai; ELISA kits for T 3 and T 4 estimations were purchased from Rapid Diagnostic Pvt. Ltd., Delhi, India. For the estimation of serum glucose and different lipids, specific kits were obtained from Span Diagnostic Limited, India. All other chemicals were obtained from Sisco Research Laboratories Pvt. Ltd., Mumbai, India.
6
Thyroid Hormone Regulation in Rats
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Adult male Wistar rats aged 11 -13 weeks and weighing 250 -320 g, bred in the Instituto de Medicina y Biología Experimental de Cuyo (IMBECU)-CONICET, were maintained on a 14/10-h light/dark cycle in a temperature-controlled environment (22 ± 2°C) with ad-libitum access to standard rat chow and water. The procedures performed in animals were consistent with the standards established by the National Institutes of Health Guide for the Care and Use of Laboratory Animals (2011) and the American Veterinarian Medical Association Guidelines on Euthanasia. A total of 48 animals were divided into control and treated. Control (euthyroid): maintained in the same conditions of temperature and humidity as the treated ones (n=16); hypoT (hypothyroidism): received 6-propyl-2-thiouracil (PTU, Sigma) at a concentration of 0.1 g/l administered in the drinking water for 21 days (n=16); hyperT (hyperthyroidism): received L-thyroxine (L-T4, Sigma) 250 μg/kg administered subcutaneously for 21 days (n=16). We have previously shown that these treatment regimens induce changes in T3, T4 and TSH consistent with hypo and hyperthyroid states [16, 17] .
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