RESUMEN
Some pharmacologic effects on bone modeling may not be evident in studies of remodeling skeletons. This study analyzes some effects of olpadronate on cortical bone modeling and post-yield properties in femurs diaphyses (virtually only-modeling bones) of young rats by mid-diaphyseal pQCT scans and bending tests. We studied 20/22 male/female animals traetad orally with olpadronate (45-90 mg/kg/d, 3 months) and 8/9 untreated controls. Both OPD doses enhanced diaphyseal cross-sectional moments of inertia (CSMI) with no change in cortical vBMD and elastic modulus. Yield stiffness and strength were mildly increased. Post-yield strength, deflection and energy absorption were strikingly enhanced. Ultimate strength was enhanced mainly because of effects on bone mass/geometry and post-yield properties. The large improvement of post-yield properties could be explained by improvements in bone geometry. Improvements in bone mass/geometry over weight-bearing needs suggest an enhanced modeling-related response to mechanical stimuli. Effects on tissue microstructural factors (not measured) could not be excluded. Results reveal novel olpadronate effects on bone strength and toughness unrelated to tissue mineralization and stiffness, even at high doses. Further studies could establish whether this could also occur in modeling-remodeling skeletons. If so, they could counteract the negative impact of anti-remodeling effects of bisphosphonates on bone strength.
Asunto(s)
Conservadores de la Densidad Ósea/farmacología , Remodelación Ósea/efectos de los fármacos , Calcificación Fisiológica/efectos de los fármacos , Difosfonatos/farmacología , Análisis de Varianza , Animales , Fenómenos Biomecánicos , Densidad Ósea/efectos de los fármacos , Diáfisis/anatomía & histología , Diáfisis/fisiología , Relación Dosis-Respuesta a Droga , Módulo de Elasticidad , Elasticidad , Femenino , Fémur/anatomía & histología , Fémur/fisiología , Masculino , Ratas , Ratas Wistar , Caracteres Sexuales , Programas Informáticos , TomografíaRESUMEN
The purpose of this study was to investigate if specific immune responses were present in mice bearing a lung adenocarcinoma that presents paraneoplastic syndromes during tumor evolution. Leukocytosis, mainly due to polymorphonuclear leukocytes, was found from day 15 of tumor growth. Delayed type hypersensitivity response and increased interleukin-6 (IL-6) serum levels were observed along tumor growth. Concomitant immunity, specific rejection of a second inoculum and in vitro specific cytotoxicity occurred at 20 days of implant. In advanced stages of tumor evolution impaired cytotoxicity, accompanied by a great increase of IL-6 in serum, were observed. Role of polymorphonuclear leukocytes and IL-6 overproduction as responsible for immune dysregulation and paraneoplastic syndromes are discussed.
Asunto(s)
Adenocarcinoma/inmunología , Neoplasias Pulmonares/inmunología , Síndromes Paraneoplásicos/inmunología , Adenocarcinoma/sangre , Animales , División Celular , Hipersensibilidad Tardía/inmunología , Inmunidad Celular , Inmunización , Interleucina-6/sangre , Recuento de Leucocitos , Neoplasias Pulmonares/sangre , Linfocitos/fisiología , Ratones , Ratones Endogámicos BALB C , Trasplante de Neoplasias , Síndromes Paraneoplásicos/sangre , Bazo/patología , Células Tumorales CultivadasRESUMEN
Paraneoplastic syndromes are rarely described in animal models. It may be useful to have a suitable experimental model to study the mechanisms by which they are produced. In this study, we describe a murine lung adenocarcinoma, P07, which presents hypercalcemia, leukocytosis and cachexia. We determined the presence of PTHrP in plasma as well as GM-CSF produced by P07 cells. TNF-alpha, which is responsible for cachexia, could neither be detected in serum nor in P07 cell supernatants. We conclude that this model, which shows paraneoplastic syndromes similar to those of lung tumor patients, should be useful to study the pathways and significance of these signs.
Asunto(s)
Adenocarcinoma/fisiopatología , Neoplasias Pulmonares/fisiopatología , Síndromes Paraneoplásicos/fisiopatología , Adenocarcinoma/sangre , Adenocarcinoma/patología , Alanina Transaminasa/sangre , Fosfatasa Alcalina/sangre , Animales , Aspartato Aminotransferasas/sangre , Proteínas Sanguíneas/análisis , Caquexia/fisiopatología , Calcio/sangre , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/biosíntesis , Hematócrito , Recuento de Leucocitos , Neoplasias Pulmonares/sangre , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos BALB C , Síndromes Paraneoplásicos/sangre , Proteína Relacionada con la Hormona Paratiroidea , Proteínas/análisis , Células Tumorales CultivadasRESUMEN
Ovariectomy and immobilization in rats have demonstrated to be useful models for osteopenia and they are considered to mimic some aspects of human osteoporosis associated with a deficit of ovarian hormones and the absence of mechanical function (disuse of the bone). Pamidronate (APD) and Olpadronate (OLPA), a new dimethylated aminobisphosphonate, on a continuous oral scheme (APD: 8 and OLPA: 0.8 mg/kg/day) or on an intermittent parenteral scheme (APD: 1.25 and OLPA: 0.075 mg/kg every 15 days) did effectively prevent the trabecular bone loss caused by immobilization (unilateral sciaticectomy), by lack of ovarian stimuli (bilateral ovariectomy) or by both approaches. There were no signs of deterioration in the cortical bone mass. In a model of preestablished osteopenia, caused by estrogen deprivation, OLPA stopped the progression of the bone mass loss (0.5 mg/kg/i.v. every 15 days) and restored (0.30-0.60 mg/kg/i.v. every 15 days) the bone mineral density which had been affected (trabecular and cortical). The different activity of OLPA and APD on trabecular and cortical regions of long bones seems to accompany their different responses because of negative stimulus: better responses were more evident in the trabecular bone which proved to be more labile. In these "in vivo" models of OLPA's efficacy was similar to APD's but it was roughly 5-10 times more potent. OLPA has a high safety margin. Therefore, it could advantageously be used in those bone diseases which benefit with the use of bisphosphonates.
Asunto(s)
Densidad Ósea/efectos de los fármacos , Enfermedades Óseas Metabólicas/tratamiento farmacológico , Huesos/efectos de los fármacos , Difosfonatos/farmacología , Análisis de Varianza , Animales , Enfermedades Óseas Metabólicas/metabolismo , Huesos/metabolismo , Difosfonatos/metabolismo , Femenino , Masculino , Ovariectomía , Pamidronato , Ratas , Ratas WistarRESUMEN
Bone mechanical competence (stiffness, strength) at organ level is determined by mechanical quality (intrinsic stiffness) and spatial distribution (macro-architecture) of bone material in cortical tissue (in every bone) and trabecular network (in vertebral bodies). These properties are inter-related and controlled according to mechanical usage by a feed-back mechanism known as mechanostat. Therefore, the effects on bone fragility of any treatment should be evaluated concerning the way they may have affected bone material or geometric properties as well as the mechanostatical interactions between them. Standard densitometry does not provide the necessary data, but some alternative methodologies (as peripheral quantitative computed tomography, pQCT) are being developed to complement or even substitute SPA, DPA or DXA determinations. Bisphosphonate (BP) effects on bone biomechanics have been studied only in animal models. Many sources of variation of results (type of compound, dose, mode of administration, species, race, sex, age, age since menopause, type of bone, remodeling ability of the skeleton, endocrine-metabolic status, interactions with other treatments, etc.) have been reported. In general terms, BPs are beneficial concerning cortical bone strength in purely modeling species (rodents) and trabecular strength in remodeling mammals (dogs, baboons). This positive action at organ level depends on independent improvements in bone macro-architecture (mainly affected by bone modeling) and material stiffness (chiefly affected by bone composition and remodeling). On one hand, bone macro-architecture has been positively affected by BPs in normal (not in ovariectomy (OX), steroid- or disuse-induced osteopenic) animals. On the other, bone material quality has been improved in the latter but not in the former. Mechanostatic interrelationships have been differently affected according to the compound employed. Results reported by ours and other laboratories concerning the three derivatives available nowadays in Argentina were reviewed and summarized. Pamidronate improved small rodents' cortical bone strength and geometric properties at low doses but impaired mineralization, material properties and strength at toxic doses. In normal, remodeling animals it improved mechanical properties in vertebral bodies but not in long bones. It also prevented the negative impact of OX-, steroid- or disuse-induced osteopenia in rats by improving bone material properties without affecting normal mechanostatic interrelationships. Olpadronate exerted positive effects on long-bone strength at any dose in normal rats and mice by improving cross-sectional properties and preserving both mineralization and material properties. These effects were highly dependent upon bone deformability, body weight, and mechanical usage of the limb as an evidence of an anabolic interaction induced on bone modeling and mechanostatic interrelationships. This compound also prevented the OX- or disuse-induced impairment in rat cortical long-bone strength and recovered rat cortical bone when given since 3 months after OX by improving only bone material quality. No interaction with bone mechanostat was detected in these studies. Alendronate effects on bone biomechanics in normal rats and dogs were positive only in long treatments. They were highly dependent on body weight of the animals, hence a positive interaction with bone mechanostat should be hypothesized. It also prevented the negative impact of OX in rat femurs by improving cortical material quality with no effect on cross-sectional properties, i.e., exerting an anti-catabolic interaction with bone mechanostat. The effects of all the three compounds were found positive for bone health, yet their mechanisms of action varied with type of bone and subject condition. A striking dissociation between (positive) effects on bone strength and (variable) effects on bone stiffness was repeatedly observed in these studies. Also an enla
Asunto(s)
Enfermedades Óseas Metabólicas/tratamiento farmacológico , Enfermedades Óseas Metabólicas/fisiopatología , Huesos/efectos de los fármacos , Huesos/fisiopatología , Difosfonatos/farmacología , Difosfonatos/uso terapéutico , Animales , Fenómenos Biomecánicos , Densidad Ósea/efectos de los fármacos , Remodelación Ósea/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , RatasRESUMEN
Bisphosphonates regulate bone turnover by inhibiting osteoclastic bone resorption. Due to their pharmacodynamic and pharmacokinetic characteristics, bisphosphonates have a special pharmacotoxicological profile related to their high degree of specificity: low or non-existent distribution in soft tissues and strong affinity for calcified tissues. Some general conclusions may be drawn from the pre-clinical toxicological studies, whose main aim is to identify the toxicity target organ/s and estimate the safety margins of a "prospective therapeutic agent" in laboratory animals. They are based on our own results and on data from the available literature as regards various bisphosphonates: Alendronate, Clodronate, Etidronate, Olpadronate and Pamidronate. Generally, very high doses of bisphosphonates are required to produce in different levels and incidence various extra-skeletical toxic side effects: local reaction, hypocalcemia (and its consequences on the cardiovascular system and the possibility of tetany), affection of the dental structures and renal dysfunction. Most of side effects may be related to the low solubility in biological fluids, the formation of calcium complexes, the potent inhibitory effect of endogenous or induced bone resorption as well as to its main excretion pathway. Some other side effects (on the eye, lungs and liver), may be related to repeated excessive high doses. A safety margin of 200 to 300 : 1 between the "toxic" and "pharmacological" doses may be estimated if the total quantity of Olpadronate given to various animal species in toxicological studies and in pharmacodynamic experimental models (osteopenias due to estrogen deprivation or immobilization and retinoid-induced hypercalcemia) is considered. If the toxic doses in animals are related to the highest doses suggested for human beings, then the ratio increases from 300 to 1000 : 1 depending on the pathology and the route of administration. As regards their effect on the bone, experimental data with the new bisphosphonates suggest a significant dissociation between pharmacologically active doses and those ones producing defective mineralization. The excessive inhibition of bone remodelling, due to the use of high doses in normal animals, is the natural consequence of the pharmacological effect of this family of compounds. A bisphosphonate's toxic potential effect on bone should not be evaluated in normal animals but in particular situations with a high bone turnover. Furthermore, the doses should be adjusted in order to regulate the magnitude of bone remodelling inhibition so as to take it to a normal level without totally suppressing it. Potency, safety margins, doses and proper administration schemes, should be considered as key elements for the optimum use of the therapeutic potentiality of these compounds.
Asunto(s)
Densidad Ósea/efectos de los fármacos , Remodelación Ósea/efectos de los fármacos , Huesos/efectos de los fármacos , Sistema Digestivo/efectos de los fármacos , Difosfonatos/toxicidad , Animales , Difosfonatos/administración & dosificación , Difosfonatos/farmacología , Evaluación Preclínica de Medicamentos , Humanos , Hipocalcemia/inducido químicamente , Ratones , Conejos , RatasRESUMEN
Female Wistar rats aged 3 months were ovariectomized (OX, n = 27). Three months later they were given i.v. doses of 150 (6), 300 (7), or 600 (6) ug/kg 2/wk of olpadronate during 12 weeks or left as OX controls (OXc). Bending fracture load of femur diaphyses, reduced in OXc, was recovered by olpadronate. This effect was paralleled by changes in material quality indicators as DEXA-BMD, tomographic (volumetric) BMD, elastic modulus, and maximum elastic stress of cortical bone. No changes were induced by any of the treatments on cross-sectional area or moment of inertia. Diaphyseal stiffness, not reduced by OX, was enhanced to overnormal values by olpadronate at any dose. None of the treatments affected the normal mechanostatic interrelationships between cross-sectional architecture and bone material quality indicators. The positive effects described point out important differences in bisphosphonate action on bone biomechanics according to the experimental conditions assayed.
Asunto(s)
Absorciometría de Fotón , Huesos/efectos de los fármacos , Difosfonatos/farmacología , Ovario/fisiología , Tomografía Computarizada por Rayos X , Animales , Fenómenos Biomecánicos , Femenino , Inyecciones Intravenosas , Ovariectomía , Ratas , Ratas Wistar , Resistencia a la Tracción/efectos de los fármacosRESUMEN
As part of a safety-assessment study, doses of 8, 40, and 200 mg/kg per day, 6 days per week, of sodium olpadronate (dimethyl-APD, Me2-APD) were given by gavage to 10-week-old male and female rats during 27 weeks. Only the 200 mg/kg per day dose provoked toxic effects and a meaningful growth depression, regardless of the animal gender. In male animals, doses of 40 or 200 mg/kg per day improved strength, stiffness, and cross-sectional moment of inertia (CSMI) of femur diaphyses despite the toxic effects observed at the highest dose. Changes in bone mechanical properties were a consequence of those induced in CSMI. Regression analyses showed a treatment-induced improvement in bone modeling (as assessed by CSMI) for the same level of bone material stiffness (as expressed by calculated values of elastic modulus). The high dependency of results on body mass bearing suggested that these effects were exerted through an increase in the efficiency of bone mechanostat. Strikingly, they were not evident in female rats. If not related to a lower bone bioavailability of bisphosphonates in female rats as described by others, this phenomenon may have reflected: (1) their a smaller biomass; and/or (2) a less effective mechanostatic regulation of bone architecture derived from a higher bone material stiffness related to male animals. An increase of BMD with a predominance toward the distal region was observed in all femurs studied. This effect, unrelated to the observed changes in mechanical properties, seems to express a lack of remodeling of primary cartilage or bone tissue.
Asunto(s)
Densidad Ósea/efectos de los fármacos , Difosfonatos/toxicidad , Fémur/efectos de los fármacos , Absorciometría de Fotón , Análisis de Varianza , Animales , Disponibilidad Biológica , Fenómenos Biomecánicos , Peso Corporal/efectos de los fármacos , Remodelación Ósea/efectos de los fármacos , Difosfonatos/administración & dosificación , Femenino , Fémur/ultraestructura , Masculino , Distribución Aleatoria , Ratas , Ratas Wistar , Análisis de Regresión , Factores Sexuales , Programas InformáticosRESUMEN
A compensation for differences in bone material quality by bone geometric properties in femora from two different strains of rats was previously shown by us. A feedback mechanism controlling the mechanical properties of the integrated bones was then proposed, in accordance with Frost's mechanostat theory. Evidence of such a system is now offered by the finding of a negative correlation between the modeling-dependent cross-sectional architecture (moment of inertia) and the mineral-dependent stiffness (elastic modulus) of bone material in the femoral diaphyses of 45 normal Wistar rats of different sexes, ages, and sizes. The strength and stiffness of the integrated diaphyses were found to depend on both cross-sectional inertia and body weight, not on bone mineral density. These findings are interpreted as supporting the hypothesis that the architectural efficiency of diaphyseal cross-sectional design resulting from the spatial orientation of bone modeling during growth is optimized as a function of the body weight-dependent bone strain history, within the constraints imposed by bone stiffness. Results suggest a modulating role of biomass, related to the system set point determination, and explain the usually observed lack of a direct correlation between mineral density and strength or stiffness of long bones in studies of geometrically inhomogeneous populations.
Asunto(s)
Peso Corporal , Densidad Ósea , Fémur/fisiología , Soporte de Peso/fisiología , Animales , Femenino , Masculino , Ratas , Ratas WistarRESUMEN
Femur diaphyses of male and female Wistar rats were densitometrically and biomechanically assayed. The BMD-dependent material properties were better in female than in male bones, but cross-section geometric properties were better in male femurs. As a result, mechanical properties of the integrated diaphyses were better in males, but differences disappeared after statistical adjustment of data to a common body weight. Results evidence a feed-back mechanism locally controlling the strain-dependent bone modelling and the corresponding cross-sectional design as related to bone stiffness, with a set-point adjusted to animal biomass. A sexual dimorphism of bone biomechanics is also described for the species.
Asunto(s)
Peso Corporal/fisiología , Fémur/anatomía & histología , Caracteres Sexuales , Animales , Fenómenos Biomecánicos , Densitometría , Elasticidad , Femenino , Masculino , Ratas , Ratas WistarRESUMEN
A series of 1-alkylsulfonylaryl-1,4-benzodiazepine derivatives were synthesized and assayed for their pharmacological profile. All the compounds tested exhibited a competitive antagonism of 3H-diazepam binding in cerebellum, cerebrum and submaxillary gland. Compound II (rec. INN tolufazepam) had a Ki of 12.7 nM in cerebrum and 400 nM in the submaxillary gland. It was very potent in preventing convulsions elicited by pentylenetetrazol (ED50 p.o.: 16.5 and ED50 i.v.: 20 mg/kg). This anticonvulsant action was suppressed by previous administration of Ro 15-1788. Compound II was also active in inhibiting suppressive behaviour in the test of Vogel. This compound has a relative low hypnogenic activity as well as a low potency to produce motor incoordination. Our results show that tolufazepam has a potential clinical usefulness.