Wistar Rat Fetuses Research Articles

An advanced ultrasound-activated drug delivery system with piezocatalytic MoS2 nanoflowers for treating patent ductus arteriosus

Patent ductus arteriosus (PDA) is a prevalent cardiovascular anomaly that leads to considerable morbidity and mortality in premature infants. Although pharmacological interventions are commonly used for treatment, they often have notable adverse effects. This study introduces an innovative approach: a controlled drug delivery system that utilizes piezocatalyst nanocarriers produced through hydrothermal techniques. The synthesized nanoparticles undergo thorough characterization to assess their structural and morphological properties. Through ultrasound treatment, precise drug loading and efficient release of indomethacin (IDM) from chitosan (CS) stabilized molybdenum disulfide nanoflowers (MoS2-CS-IDM) are achieved. By harnessing ultrasound-induced reactive oxygen species, this method enables targeted drug delivery. Importantly, the nanoparticles demonstrate an approximately 60 % decrease in PGE2 concentrations, indicating their potential as anti-inflammatory agents. The favorable interaction of the nanoparticles with rat aortic smooth muscle cells and the successful closure of the ductus arteriosus in fetal Wistar rats validate their therapeutic effectiveness in vivo. In conclusion, the piezocatalyst-based drug delivery system presented in this study holds promise for versatile, biocompatible, and controlled drug release, with significant implications for therapeutic applications in PDA and potentially beyond.

Pathomorphological characteristics of heat stress in the experiment.

The purpose of this research was to explore some morphological, physiological, and biochemical changes in female and fetal Wistar rats under heat stress. The experiment involved 30 animals, including two experimental groups (pregnant and nonpregnant females) kept under heat stress at 32°C and one control group consisting of healthy individuals kept in standard vivarium conditions. After dissection, fixation, dehydration, and primary processing, tissue samples were embedded in a mixture of paraffin and lanolin to obtain material for sections. Sections were made using a freezing and angular microtome and stained with hematoxylin and fuchsine solutions. Changes in morphology were assessed by microscopy using a Leitz DIAPLAN system. As a result of heat stress, an increase in linear cell size, capillary network area, and adrenal mass was observed; adipocytes lost lipid vacuoles; prismatic thyroid cells were replaced by flat cells; hypothyroidism; an increase in the number of osteocyte lacunae; and increased osteoclast activity in bone tissue; interstitial and intracellular oedema and caryopycnosis of ventricular cardiomyocytes; reduction in the diameter of skeletal muscle fibers and replacement of tissue with collagen fibers; water loss in the structure of myofibrils; destructive local changes, hyperchromatosis and caryopycnosis of the hippocampus. The data obtained allows predicting the possible consequences of prolonged overheating of tissues of other vertebrates and the human body.

Immunohistochemical Characterization of Heart Left Ventricle Morphogenesis in Rats Fetuses

Rats are the most common objects of preclinical studies, which determines the relevance of studying fetal cardiomorphogenesis in rats using modern methods of morphological studies. The aim is to study the dynamics of markers of proliferation (Ki67), apoptosis (caspase 3), vascularization (CD31), and stromal remodeling (MMP2 and MMP9) of the left ventricular myocardium of Wistar rat fetuses. Material and methods. A histological, immunohistochemical and morphometric study of the rat heart left ventricle wall was carried out on the 18th, 19th, 20th, 21st and 22nd days of the prenatal period of ontogenesis. Results. From the 18th to the 22nd day of the prenatal period of ontogenesis in rats, a twofold increase in the thickness of the left ventricle lateral wall is observed, due to both proliferation and differentiation and an increase in the size of cardiomyocytes. Ki67-positive cardiomyocytes are diffusely localized in the wall of the left ventricle, their number increases on day 19 of the prenatal period of development and remains at a high level until the end of the prenatal period in rats. From the 18th to the 22nd day of the prenatal period of ontogeny in rats, single caspase 3-positive cardiomyocytes are detected in the left ventricle. The growth of the left ventricular myocardium in the fetal period in rats is accompanied by an adequate rate of vascularization. The vascularization and proliferation of cardiomyocytes are accompanied by remodeling of the myocardial stroma, with the highest intensity of MMP2- and MMP9-immune staining observed at the beginning of the fetal period of ontogeny in rats. Conclusion. The short duration of prenatal ontogenesis in rats determines the high dynamics of cardiomorphogenetic processes. In the fetal period of rat ontogenesis in the left ventricle of the heart, the proliferation of cardiomyocytes prevails over the intensity of apoptosis. The intensity of proliferation of cardiomyocytes of the left ventricle of rats is high up to the 22nd day of the prenatal period of ontogenesis.

Intrauterine desensitization enables long term survival of human oligodendrocyte progenitor cells without immunosuppression

Immune rejection can be reduced using immunosuppressants which are not viable for premature infants. However, desensitization can induce immune tolerance for premature infants because of underdeveloped immune system. The fetuses of Wistar rats at 15-17days gestation were injected via hOPCs-1 into brain, muscles, and abdomen ex utero and then returned while the fetuses of control without injection. After 6weeks of desensitization, the brain and muscles were transplanted with hOPCs-1, hNSCs-1, and hOPCs-2. After 10 and 34weeks of desensitization, hOPCs-1 and hNSCs-1 in desensitized groups was higher than that in the control group while hOPCs-2 were rejected. Treg, CD4CD28, CD8CD28, and CD45RC between the desensitization and the control group differed significantly. Inflammatory cells in group with hOPCs-1 and hNSCs-1 was lower than that in the control group. hOPCs-1 can differentiate into myelin in desensitized groups. Wistar rats with desensitization developed immune tolerance to desensitized and transplanted cells.

Effect of preconditioning on propofol-induced neurotoxicity during the developmental period.

At therapeutic concentrations, propofol (PPF), an anesthetic agent, significantly elevates intracellular calcium concentration ([Ca2 +]i) and induces neural death during the developmental period. Preconditioning enables specialized tissues to tolerate major insults better compared with tissues that have already been exposed to sublethal insults. Here, we investigated whether the neurotoxicity induced by clinical concentrations of PPF could be alleviated by prior exposure to sublethal amounts of PPF. Cortical neurons from embryonic day (E) 17 Wistar rat fetuses were cultured in vitro, and on day in vitro (DIV) 2, the cells were preconditioned by exposure to PPF (PPF-PC) at either 100 nM or 1 μM for 24 h. For morphological observations, cells were exposed to clinical concentrations of PPF (10 μM or 100 μM) for 24 h and the survival ratio (SR) was calculated. Calcium imaging revealed significant PPF-induced [Ca2+]i elevation in cells on DIV 4 regardless of PPF-PC. Additionally, PPF-PC did not alleviate neural cell death induced by PPF under any condition. Our findings indicate that PPF-PC does not alleviate PPF-induced neurotoxicity during the developmental period.

Teratogenic Effect of Aqueous Leaf Extract of Aspilia africana on The Dentate Gyrus of Albino Wistar Rat Fetuses

Aspilia africana is an herbal plant widespread in Africa used for medicinal purposes and also used by pregnant women for health related issues. This study was aimed at investigating the teratogenic effect of aqueous leaf extract of Aspilia africana on the dentate gyrus of albino wistar rat fetuses. Twenty (20) female adult rats weighing between 190-205g were used for this study. The rats were divided into four groups; control, low dose, medium dose and high dose with each group containing five rats. Pregnancy was induced by caging the female rats with sexually matured males. The presence of vaginal plug and tail structure in the vaginal smear the following morning confirmed coition, and it was regarded as day 0 of pregnancy. The control group was given distilled water. The low dose, medium dose, and the high dose groups received 750mg/kg, 1000mg/kg, and 1250mg/kg body weight of aqueous leaf extract of Aspilia africana through an orogastric tube from day 7-11 of gestation. On the 20th day of gestation, the animals were sacrificed using chloroform-inhalation method. Their fetuses were harvested via uterectomy, the brain was excised and fixed in 10% buffered formalin, and then routine histological processes were carried out. Staining was done using Haematoxylin and Eosin method. Histological observation of the dentate gyri of experimental groups revealed marked distortion, reduction of the polymorphic layer, hyperplasia and hypertrophy of cells in the molecular and granular layer especially in the high dose group whose mothers received 1250mg/kg of the extracts. The result suggests high doses of aqueous leaf extract of Aspilia africana may be teratogenic to the dentate gyrus of Wistar rat fetuses.

Teratogenic Effect of Aqueous Leaf Extract of Aspilia africana on The Dentate Gyrus of Wistar Rat Fetuses

Aspilia africana is an herbal plant widespread in Africa used for medicinal purposes and also used by pregnant women for health related issues. This study was aimed at investigating the teratogenic effect of aqueous leaf extract of Aspilia africana on the dentate gyrus of albino wistar rat fetuses. Twenty (20) female adult rats weighing between 190-205g were used for this study. The rats were divided into four groups; control, low dose, medium dose and high dose with each group containing five rats. Pregnancy was induced by caging the female rats with sexually matured males. The presence of vaginal plug and tail structure in the vaginal smear the following morning confirmed coition, and it was regarded as day 0 of pregnancy. The control group was given distilled water. The low dose, medium dose, and the high dose groups received 750mg/kg, 1000mg/kg, and 1250mg/kg body weight of aqueous leaf extract of Aspilia africana through an orogastric tube from day 7-11 of gestation. On the 20th day of gestation, the animals were sacrificed using chloroform-inhalation method. Their fetuses were harvested via uterectomy, the brain was excised and fixed in 10% buffered formalin, and then routine histological processes were carried out. Staining was done using Haematoxylin and Eosin method. Histological observation of the dentate gyri of experimental groups revealed marked distortion, reduction of the polymorphic layer, hyperplasia and hypertrophy of cells in the molecular and granular layer especially in the high dose group whose mothers received 1250mg/kg of the extracts. The result suggests high doses of aqueous leaf extract of Aspilia africana may be teratogenic to the dentate gyrus of Wistar rat fetuses.

Skeletal malformations and growth disturbances in fetuses of mild diabetic rats

ResumenIntroducción. En la actualidad, la diabetes mellitus representa una de las condiciones médicas que complica el embarazo con mayor frecuencia, lo que afecta el crecimiento y el desarrollo fetal.Objetivo. Determinar las malformaciones esqueléticas y alteraciones en el crecimiento en fetos de ratas Wistar diabéticas.Materiales y métodos. Se utilizó un modelo de diabetes moderada inducida neonatalmente con estreptozotocina (STZ 100 mg/kg de peso corporal, por vía subcutánea) en ratas Wistar. En la adultez, las ratas sanas y diabéticas se aparearon con machos sanos de la misma edad y cepa. El día 20 de gestación se practicó la cesárea bajo anestesia. Se extrajeron los fetos, se pesaron y clasificaron como pequeños (PAG), adecuados (AEG) o grandes (GEG) para la edad gestacional. Los fetos seleccionados se procesaron para el análisis de anomalías esqueléticas y sitios de osificación. Resultados. En la descendencia de las ratas diabéticas, hubo un mayor porcentaje de fetos clasificados como pequeños o grandes y un menor porcentaje de fetos con peso adecuado; el promedio de peso fetal fue menor y había menos sitios de osificación. Se observaron alteraciones en la osificación de cráneo, esternón, columna vertebral, costillas y extremidades anteriores y posteriores; y también, hubo una correlación directa entre el peso y el grado de osificación fetal.Hubo malformaciones congénitas asociadas con la fusión y bifurcación de las costillas, así como cambios indicativos de hidrocefalia, como la forma de domo del cráneo, una amplia distancia entre los parietales y la anchura de las fontanelas anterior y posterior. Conclusión. La diabetes moderada durante la gestación altera el crecimiento y el desarrollo fetal, que se ve afectado tanto por macrosomía y la restricción del crecimiento intrauterino como por malformaciones esqueléticas.

ВЛИЯНИЕ ПРОМЫШЛЕННОЙ ВИБРАЦИИ НА МИНЕРАЛЬНЫЙ ОБМЕН, ОРГАНЫ ПОЛОСТИ РТА И МИОКАРД ПЛОДА

In the study, the morphometric analysis of bone trabeculae of the developing jaws, tooth buds, tongue muscles, masseter muscle, and myocardium of 20-day fetuses of Wistar rats, exposed to vibration from the 9th to the 18th day of prenatal ontogenesis, was performed. The concentration of Ca, Cd, Cu, Fe, Mg, P, Pb, Se, Zn was determined in the liver of pregnant females exposed to vibration. The morphometric study revealed an acceleration of the osteogenesis of jaws and dentinogenesis of tooth buds. In the myocardium and masseter muscle signs of interstitial edema and a decrease in the area of the vessels of the microvasculature are determined. In the liver, there is a decrease in the amount of Ca, Mg and Fe, which is accompanied by an increase in the content of Cd and Pb. The revealed shifts in mineral metabolism indicate gross impairment of chemicals' homeostasis in the mother-fetus system, which underlies the imperfect morphogenesis of the fetal dentition and will be the basis for the formation of pathology of orofacial organs and cardiovascular pathology in offspring.

Kolaviron ameliorates toxic effects of aluminum chloride on the hippocampus of fetal Wistar rats in utero: Biochemical and ultrastructural observations

BACKGROUND: This study was designed to investigate the biochemical and ultrastructural effects of kolaviron (Kv) on the hippocampus of fetal Wistar rats exposed to aluminum chloride (AlCl3) toxicity in utero. MATERIALS AND METHODS: Fifty female Wistar rats were selected at random and mated. Following confirmation of mating, pregnant rats were assigned into five groups (n = 5). Controls: Group A received distilled water; Group B: 0.6 ml of corn oil; Group C: 200 mg/kg of Kv; Group D: 100 mg/kg of AlCl3 and Group E 100 mg/kg of AlCl3 and 200 mg/kg of Kv. Administration was done from days 8-10 and 15-17 of gestation during the 2nd and 3rd weeks respectively. Biochemical analyses were investigated to assess oxidative stress levels, while transmission electron microscopy (TEM) examined ultrastructural changes. Pregnant animals were sacrificed on day 20 of gestation; fetuses, their brains, and hippocampi were excised, respectively. Hippocampal tissues of fetuses were homogenized in 0.25 M of sucrose solution for biochemical assay while some were fixed in 2.5% phosphate-buffered saline-based glutaraldehyde for TEM. RESULTS: Elevated levels of Al, malondialdehyde, nitric oxide, and interleukin 6 were observed in the hippocampi of fetuses whose mothers received AlCl3. TEM revealed loss of nuclear membrane and increased condensation of chromatin materials in the same group. However, significant reduction of these enzymes including improved ultrastructural alterations were observed in the fetal hippocampus of the AlCl3 + Kv-treated group. CONCLUSION: This study showed that Kv significantly reduced neurodegenerative effects induced by AlCl3 in the hippocampii of fetal Wistar rats in utero probably owing to its antioxidant and anti-inflammatory properties.

THE EFFECT OF LIMAN LEAVES EXTRACT ON FETAL RAT DEVELOPMENT

Objective: The objective of the study was to observe the effect of liman leaves extract on fetal rat development and provide safety information for its use during pregnancy.
 Methods: Estrus cycles of female Wistar rats were observed and mated to male rats, 0 day of pregnancy determined after finding a vaginal plug or in a vaginal smear, there was sperm. Sample was administered orally using sonde with a single administration on the 11th day of pregnancy, at doses of 3750 mg/kg bw, 1185 mg/kg bw, and 375 mg/kg bw. On the 19th day of pregnancy, the rats were sacrificed and then observed the number of implantation, corpus luteum, intrauterine death, and fetal abnormalities.
 Results: The dose of 375 mg/kg bw exposed the highest average implantation rate (13.20%) and the largest number of corpus luteum (13.90%). The highest total intrauterine death was presented by dose of 3750 mg/kg bw and significantly different (p<0.05) compared to the control group. The dose of 375 mg/kg bw expressed the highest percentage of embryos with resorption (19.03%), while the lowest average fetal body was shown by 3750 mg/kg bw compared to the other groups. The highest percentage of external abnormalities was given by the dose of 375 mg/kg bw (12.91%), which abnormalities found were dwarf, cleft palate, hydrocephalus, short sleeve, and hematoma.
 Conclusion: Liman leaves extract was mild teratogenic on Wistar rat fetus.

The ossification status of Wistar rat fetuses at the end of gestation

We present a comprehensive description of spontaneous skeletal development of Wistar rat fetuses close to parturition. Fetuses were collected on the day of expected parturition and one and two days before (Gestation Days 21–23) and skeletal development state was examined. Data is presented on an individual fetal basis to allow insights in developmental patterns. The distal parts of the limbs and the cervical and caudal vertebral column were identified as areas of the greatest developmental difference in the examined time frame. Extensive knowledge of the ossification process in individual fetuses allows a better resolution of deviations from normal and gives valuable information for the grading of delayed maturation and structural deviations. The data base is helpful to overcome the bottleneck of coincident occurrence of low incidences of skeletal abnormalities in the dose groups and the control group complicating risk assessment of test articles.

Influence of Adriblastin and Bleomycin on Wistar rat mothers and fetus development

Influence of Adriblastin and Bleomycin on Wistar rat mothers and fetus development

Fas- and Mitochondria-Mediated Signaling Pathway Involved in Osteoblast Apoptosis Induced by AlCl3.

Aluminum (Al) is known to induce apoptosis of osteoblasts (OBs). However, the mechanism is not yet established. To investigate the apoptotic mechanism of OBs induced by aluminum trichloride (AlCl3), the primary OBs from the craniums of fetal Wistar rats were exposed to 0mg/mL (control group, CG), 0.06mg/mL (low-dose group, LG), 0.12mg/mL (mid-dose group, MG), and 0.24mg/mL (high-dose group, HG) AlCl3 for 24h, respectively. We observed that AlCl3 induced OB apoptosis with the appearance of apoptotic morphology and increase of apoptosis rate. Additionally, AlCl3 treatment activated mitochondrial-mediated signaling pathway, accompanied by mitochondrial membrane potential (ΔΨm) depolarization, release of cytochrome c from the mitochondria to the cytoplasm, as well as survival signal-related factor caspase-9 and caspase-3 activation. AlCl3 exposure also activated Fas/Fas ligand signaling pathway, presented as Fas, Fas ligand, and Fas-associated death domain expression enhancement and caspase-8 activation, as well as the hydrolysis of Bid to truncated Bid, suggesting that the Fas-mediated signaling pathway might aggravate mitochondria-mediated OB apoptosis through hydrolyzing Bid. Furthermore, AlCl3 exposure inhibited Bcl-2 protein expression and increased the expressions of Bax, Bak, and Bim in varying degrees. These results indicated that AlCl3 exposure induced OB apoptosis through activating Fas- and mitochondria-mediated signaling pathway and disrupted B-cell lymphoma-2 family proteins.

Influence of gender and body hemisphere on the occurrence of wavy ribs: An analysis of spontaneous skeletal abnormalities in Wistar rat fetuses.

The differentiation of background findings from anomalies relevant for the safety evaluation of a future drug is a major task in the interpretation of developmental toxicity studies. The anomaly wavy rib often occurs in litters exposed to the test substance, but is also frequently present in control litters. Therefore, the relevance of this finding for risk assessment is under discussion. We characterized the skeletal morphology of fetuses with wavy ribs from our background data. Differences in the incidence of wavy ribs between the genders, with approximately twice as many male fetuses having wavy ribs, compared to females, and an overrepresentation of the right body hemisphere were observed. Affected fetuses often occurred clustered in single litters. The presented data might be useful for differentiation of spontaneously occurring wavy ribs from abnormal patterns induced by a test substance.

Phenotypic differentiation of neurons in intraocular transplants

Neurochemical differentiation of neurons in transplants developing in rat anterior eye chamber was studied. Pieces of the somatosensory neocortex area, isolated from 17-day fetuses of Wistar rats, were used for the transplantation. The general cytological analysis and immunochemical identification of GABAergic neurons in neocortical transplants and in the appropriate brain area of the recipient rats (control) were carried out after 6 months. Cytoarchitectonics typical for neocortex was not revealed in the transplants. Furthermore, a 1.4-fold decrease in numerical density of the entire neuron population was found compared to the control. The proportion of GABAergic nerve cells in the transplanted tissue was reduced even more dramatically— by 13.1 times. The dimensions of all types of neurons, especially GABAergic cells, were greater in the transplants in oculo compared to neocortex in situ. The increase in size occurred mostly due to the cytoplasm. Thus, the nuclei of GABA-positive neurons in the transplants were larger by 1.2 times compared to the control and their perikarya were larger by 1.5 times. The obtained results showed that the conditions in the anterior eye chamber the most dramatically affect the differentiation of GABAergic neurons, and cell hypertrophy, probably, is the functional compensation of the decrease in their number. Considering the literature data on the increased excitability and synchronized neuronal activity in the intraocular transplants, it can be assumed that these transplants can be used as a model for studying the cellular mechanisms of nervous tissue epileptization under disinhibition conditions.

Histological and Morphological Study of the Intestines of Wistar Rat Fetuses in a Modified Gastroschisis Experimental Model

In gastroschisis (G), the lesion degree of exposed intestinal segments is related to the time of its contact with the amniotic fluid (AF) and exposure to meconium which is the cause of intestinal morphological and histological alterations. The outcome of these alterations is intestinal hypoperistalsis and nutrient absorption deficiency, which contribute to increased morbidity and high medical-hospital costs. In this study, morphological and histological intestine alterations were identified at two different contact occasions with AF. Experimental gastroschisis (G) was performed on Wistar rat fetuses at a single gestational age on day 18.5th. The fetuses were removed on the 20.5th (G-1) and 21.5th days (G-2). Fetuses of both groups were divided in 3 sub-groups: control (C), gastroschisis (G) and sham (S). Measurements were taken of the Whole Set including fetus, placenta and membranes with AF (WS), fetus body weight (BW), intestinal weight (IW) and their diameters (DI). The objective of the present study is to test a new gastroschisis experimental model and identify differences in morphological and histological alterations in these two gestational periods that may be directly related to intestinal motility disorders in G. The WS and BW presented no significant statistical difference when compared G1 and G2. The results of the intestine average weight of G2 fetuses were significantly higher when compared to G1 fetuses in all subgroups (C: p = 0.02; G: p = 0.01; S: p = 0.02, Mann Whitney). The results of the intestinal average diameters (D/d) in G1 and G2 presented significant statistical difference only in G subgroup (p Kruskal Wallis). When compared intestinal average diameters, there was significant statistical difference of G fetuses in G1 and G2 (p Mann Whitney). In conclusion, the present experimental G model was adequate to reproduce G in rat fetuses. All G fetuses presented significant statistical difference when compared to other group in their subgroup and when compared G1 and G2 (p < 0.05). These alterations can explain the difficulties in accomplishing adequate peristalsis in G neonate bearers.

High Fat Diet Exposure during Fetal Life Enhances Plasma and Hepatic Omega-6 Fatty Acid Profiles in Fetal Wistar Rats.

Pregnant rats were fed a high fat diet (HFD) for the first (HF1), second (HF2), third (HF3) or all three weeks (HFG) of gestation. Maintenance on a HFD during specific periods of gestation was hypothesized to alter fetal glycemia, insulinemia, induce insulin resistance; and alter fetal plasma and hepatic fatty acid (FA) profiles. At day 20 of gestation, fetal plasma and hepatic FA profiles were determined by gas chromatography; body weight, fasting glycemia, insulinemia and the Homeostasis Model Assessment (HOMA-insulin resistance) were also determined. HF3 fetuses were heaviest concomitant with elevated glycemia and insulin resistance (p < 0.05). HFG fetuses had elevated plasma linoleic (18:2 n-6) and arachidonic (20:4 n-6) acid proportions (p < 0.05). In the liver, HF3 fetuses displayed elevated linoleic, eicosatrienoic (20:3 n-6) and arachidonic acid proportions (p < 0.05). HFG fetuses had reduced hepatic docosatrienoic acid (22:5 n-3) proportions (p < 0.05). High fat maintenance during the final week of fetal life enhances hepatic omega-6 FA profiles in fetuses concomitant with hyperglycemia and insulin resistance thereby presenting a metabolically compromised phenotype.

Impairment of bone growth of wistar rat fetuses of diabetic and hypercholesterolemic mothers

Patients with either diabetes or hypercholesterolemia develop atherosclerosis and impair healing of bone. In the present work we illustrated their role in limb ischemia during bone cells differentiation of Wistar rat fetuses. Pregnant Wistar rats (n = 20 each) were arranged in three groups: control, diabetic or hypercholesterolemic. Diabetes was induced at the fifth day of gestation using streptozotocin, and hypercholesterolemia was carried by feeding virgin rats a diet containing 3% cholesterol for 6 wks prior to the onset of conception. At 13, 15, 17, and 19 d prenatal, pregnant rats were sacrificed, dissected, and fetuses were removed. Hind limbs were separated and subjected to histological and transmission electron microscopic examination, ossification, total calcium content of fetuses, isoenzymes alkaline and acid phosphatase and lactic dehydrogenase electrophoresis and DNA damage. Fetuses of diabetic or hypercholesterolemic mothers exhibited delayed histo-cytological differentiation of chondrocytes, and decreased periosteal ossification. Alkaline and acid phosphatase as well as lactic dehydrogenase isoenzymes showed altered diffusion rate and intensities of their bands reflecting their activities in both diseases comparing with the control. Assessments of bone calcium contents revealed marked reduction. Genomic expression of the degree of laddering (total DNA fragmented) or single-cell gel electrophoresis was found to be increased in cartilage and bone cells of fetuses of diabetic or hypercholesterolemic mothers. The authors concluded that both diseases had a selective, dramatic effect during fetus development in this model by retarding the histo- and cytological differentiation during limb bone growth. Both diseases increased the average cell death in skeletal elements and blood vessels as a consequence of altered alkaline and acid phosphatases and lactic dehydrogenase isoenzymes in accordance with DNA damage.

Reduced post-natal versus pre-natal incidence of bent long bones and scapulae in a preliminary investigation using the Han Wistar rat

There is a “chondrodystrophy” syndrome in the Han Wistar rat fetus that manifests as characteristic skeletal abnormalities such as bent and/or short long bones, and is classified as permanent detrimental abnormalities (major malformations). This pilot study investigated whether these defects resolve after birth.Han Wistar rats were dosed during organogenesis either with vehicle or test article. Examination of gestation day 20 fetuses showed a slightly increased incidence (11%; 11/101) of skeletal abnormalities in the high dose fetuses compared with 6% (4/67) in control fetuses, whereas no skeletal abnormalities were present in the 205 pups examined on post-natal day 21. The probability of having zero litters containing pups with skeletal abnormalities was p<0.0000001. This very low probability suggests that these defects recover by weaning and supports the hypothesis that these fetal findings in the Han Wistar are probably not permanent abnormalities and therefore are potentially reclassifiable as minor malformations.