Effects of mechanical loads on surface morphology of the condylar cartilage of the mandible in rats

Mechanical loads on the condylar cartilage were varied by feeding either a hard diet, a soft diet, or a sequential combination of a soft diet followed by a hard diet to young male and female rats for a total period of 4 weeks, and to mature male rats for 12 weeks. Gross condylar dimensions were greatest in the hard-diet groups, intermediate in the combination soft/hard-diet groups, and smallest in the soft-diet groups. Scanning electron microscopy of the condyle revealed a smooth, nonporous articular surface in the soft-diet groups. Hard-diet condyles had a rougher, more porous articular surface while soft/hard-diet condyles were intermediate between nonporous and slightly roughened condyles. None of the condyles showed ridges or elevations on the articular surface. Sex, age and time of the diets did not significantly affect these results.     

Effect of bone strain on cortical bone structure in macaques (Macaca mulatta)

It has recently been shown that the consistency of food significantly affects levels of bone strain in the mandible during mastication (Hylander, '79a). Mandibular bone histology was examined to test the effects of a diet of hard food compared to a diet of soft food in two group of monkeys. One group of rhesus macaques (Macaca mulatta) was fed a diet of commercially prepared hard biscuits. The second group was fed a soft diet the consistency of fudge. Both diets were nutritionally adequate for normal growth and development. As a control for other factors influencing cortical bone structure, fibular morphology was also examined. At the end of the test period, mandibular and fibular tissue samples from the two groups were prepared to determine the amount of secondary Haversian bone present. Mandibular depth at M₂ and fibular anteroposterior diameter were also measured and compared between the two dietary groups. The soft-diet monkeys showed low levels of remodeling in their mandibles. There were large patches of unremodeled bone and resorption spaces were common. The hard-diet monkeys exhibited more extensive evidence of secondary Haversian remodeling in their mandibles. The hard-diet monkeys also had deeper mandibles. In contrast, the fibulae from the two groups had similar mean diameters and showed comparable levels of secondary remodeling. We infer that the higher mandibular bone remodeling levels in the hard-diet monkeys represent an adaptive response to remove and replace fatigued mandibular bone due to higher stress levels associated with the ingestion and mastication of hard foods. We also infer that greater depth of the mandible at M₂ found in the harddiet group represents an adaptive response to higher stress levels associated with the ingestion and mastication of hard foods.     

Effect of altered masticatory function on [3H]-thymidine and [35S]-sulfate incorporation in the condylar cartilage of the rat

The response of the condylar cartilage to alterations in compressive joint reaction forces in vivo has been little studied. In an attempt to reduce or eliminate the occlusal forces resulting from mastication or incision, male weanling rats were fed a soft diet requiring little chewing and/or had their incisors clipped every other day. Incorporation (dpm/micrograms DNA) of [3H]-thymidine and [35S]-sulfate was significantly decreased relative to controls in the incisor-clipped group, but not in the soft-diet group. Animals having both treatments also exhibited significantly lower incorporation values than controls, suggesting the importance of incision for loading at the mandibular joint. These data corroborate in vitro studies which suggest that compressive forces can affect mitotic activity and synthesis of proteoglycans in the condylar cartilage. However, additional factors, both hormonal and biomechanical in nature, may be important in the in vivo environment.     

Response of the mandibular joint to loss of incisal function in the rat

In a study of the rat mandibular joint (MJ), Simon [Acta anat. 97: 351-360 (1977)] suggested that reduction in condylar cartilage thickness noted in animals subjected to removal or trimming of incisors resulted from the lessening of joint reaction forces produced during incision. In order to explore this question further, the microanatomy of the MJ in 47-day-old rats whose incisors had been trimmed every other day was compared to that in control animals and in a third group fed a soft diet as a control for reduced joint reaction forces. Both the incisor-clipped and soft-diet groups exhibited reduced size and density of bony trabeculae underlying the condylar cartilage and diminished staining for alcian blue. The thickness of the prechondroblastic layer of the condylar cartilage was significantly (p less than or equal to 0.01) reduced relative to controls in both experimental groups on the superior aspect of the cartilage, but was reduced in the more posterior parts of the cartilage only in the incisor-clipped group. While not denying that joint reaction forces may affect MJ response, the reduced proliferative response noted in the posterior region of the condylar cartilage in incisor-clipped animals is perhaps best explained by a decrease in the frequency and extent of protrusion of the lower jaw due to a lack of incisal preparation of food items.     

Load application induces changes in the expression levels of Sox-9, FGFR-3 and VEGF in condylar chondrocytes

Experimental and clinical observations have proven the modulatory effects of mechanical loading on the development and maintenance of cartilage architecture. Here we examined the involvement of Sox-9, FGFR-3 and VEGF (pivotal factors controlling cartilage development and growth) in the mechano-transduction pathway of mandibular condylar cartilage by changing the dynamics of the transmitted load via changes in food hardness. To this end, condyle cartilage tissue of rats fed with hard or soft food was analyzed immunohistochemically at various time points. Our findings demonstrate that different mechanical loading conditions in condylar chondrocytes trigger differentiation-/maturation-related processes by affecting the expression levels of these factors, ultimately influencing condylar cartilage growth.     

Activities of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) on undernourished and renourished rats' thymus

We studied the effect of administration of a low quality dietary protein, from weaning onwards, on the thymus of undernourished rats and the posterior effect of refeeding with a high quality dietary protein. Changes in thymus weight and the activity of Adenosine Deaminase (ADA) and Purine Nucleoside Phosphorylase (PNP) on thymus, were determined. Wistar rats were suckled in groups of 14-16 per dam since birth to weaning (23 days) to obtain undernutrition. At weaning, a group of 14-16 rats received pre-cooked maize flour (Protein content: 6.5%) for 18 days. One group was sacrificed (M) and the other rats were refed with the casein diet (Protein content: 20%) during 20 days (R). The age-matched control groups were fed stock diet since 40 (C40) and 60 (C60) days of age, respectively. At the end of the experimental period, body (Bw) and thymus weight were determined. ADA and PNP activities were determined in thymocyte suspensions. Highly significant differences in thymus weight-expressed as mg or mg/Bw(0.75)-and the activity of ADA and PNP were observed in rats fed the experimental diet containing maize flour, when compared to the respective age-matched control. No statistical differences were observed between R and C60.The administration of a high quality dietary protein to undernourished weanling rats is capable to reverse the damage produced by the low quality dietary protein on thymus weight and ADA and PNP thymus activities.     

Hard-Diet Feeding Recovers Neurogenesis in the Subventricular Zone and Olfactory Functions of Mice Impaired by Soft-Diet Feeding

The subventricular zone (SVZ) generates an immense number of neurons even during adulthood. These neurons migrate to the olfactory bulb (OB) and differentiate into granule cells and periglomerular cells. The information broadcast by general odorants is received by the olfactory sensory neurons and transmitted to the OB. Recent studies have shown that a reduction of mastication impairs both neurogenesis in the hippocampus and brain functions. To examine these effects, we first measured the difference in Fos-immunoreactivity (Fos-ir) at the principal sensory trigeminal nucleus (Pr5), which receives intraoral touch information via the trigeminal nerve, when female adult mice ingested a hard or soft diet to explore whether soft-diet feeding could mimic impaired mastication. Ingestion of a hard diet induced greater expression of Fos-ir cells at the Pr5 than did a soft diet or no diet. Bromodeoxyuridine-immunoreactive (BrdU-ir) structures in sagittal sections of the SVZ and in the OB of mice fed a soft or hard diet were studied to explore the effects of changes in mastication on newly generated neurons. After 1 month, the density of BrdU-ir cells in the SVZ and OB was lower in the soft-diet-fed mice than in the hard-diet-fed mice. The odor preferences of individual female mice to butyric acid were tested in a Y-maze apparatus. Avoidance of butyric acid was reduced by the soft-diet feeding. We then explored the effects of the hard-diet feeding on olfactory functions and neurogenesis in the SVZ of mice impaired by soft-diet feeding. At 3 months of hard-diet feeding, avoidance of butyric acid was reversed and responses to odors and neurogenesis were recovered in the SVZ. The present results suggest that feeding with a hard diet improves neurogenesis in the SVZ, which in turn enhances olfactory function at the OB.     

Aluminum concentrations in tissues of rats: effect of soft drink packaging

Aluminum is a commonly occurring trace element for which no nutritional requirements have been set. Some non-conclusive evidence exists suggesting a need of aluminum for growth, reproduction or health of man and animals. There is concern that exposure or consumption of aluminum may be toxic to humans and animals. The objective of the current study was to compare tissue levels of aluminum of rats fed soft drinks packaged in aluminum cans, glass bottles or distilled water. Thirty male weanling rats (Sprague-Dawley) were divided into three treatment groups of 10 rats each. All rats were fed rodent chow ad libitum throughout the study. Three different fluids, i.e. distilled water, diet soft drinks from aluminum cans and diet soft drinks from glass bottles, were fed for a period of 3 weeks. Aluminum contents of tissues were measured by atomic absorption spectrophotometry. Canned soft drink fed rats had significantly higher blood, liver and bone aluminum concentration than rats that were given glass bottled soft drink. There was a 69% higher bone aluminum concentration and 16% lower femur weight in rats fed aluminum canned soft drinks when compared with rats fed with distilled water.     

Do mandibular cross‐sectional properties and dental microwear give similar dietary signals?

Previous animal experimental work evaluating the effects of dietary consistency on mastication was generally limited to studies of either mandibular structure or rates and types of tooth wear. Control groups fed hard diets (HD) consistently exhibited increased cortical remodeling and/or bone strength when compared to groups fed soft diets (SD). Results of tooth-wear studies showed faster rates of tooth wear in HD animals. This study evaluates the effects of dietary differences on both mandibular structural morphology and postcanine dental microwear in the same animals. We examined mandibles and dentitions from eight miniature swine, raised from 4 weeks to 9 months of age on HD and SD (n = 4, each group). Mandibular structural properties were calculated from peripheral quantitative computed tomography slices at the dp3-dp4 and dp4-M1 junctions. Dental microwear analysis was performed on mandibular lingual crushing facets of dp4 and M1, using photomicrographs of high-resolution casts taken at 500x magnification in a scanning electron microscope. Our results suggest that between the dp3-dp4 contact, HD animals have mandibles that are stronger and more rigid mediolaterally than SD animals. At the dp4-M1 contact, HD animals have mandibles that are stronger and more rigid mediolaterally, dorsoventrally, and in torsion than SD animals. Dental microwear results indicate that SD pigs have higher incidences of pitting and more overall microwear features on their premolars than do HD pigs, yet there are no significant differences in molar microwear morphology between the dietary groups. Near-significant correlations exist between pit size and dorsoventral bending strength, but only for HD pigs. These results suggest that dietary consistency significantly affects both mandibular structure and dental microwear, yet direct correlations between the two are complicated by a number of factors.     

Alveolar macrophage function in rats with severe protein calorie malnutrition. Arachidonic acid metabolism, cytokine release, and antimicrobial activity

To investigate the effects of protein calorie malnutrition (PCM) on alveolar macrophage function, we measured antimicrobial activity, IL-1 and TNF production, and arachidonic acid metabolism in alveolar macrophages of infant rats with moderate and severe PCM. Groups of weanling male rats were fed a diet containing 0.8% protein (PCM) or 24% protein (control). A third group (pair fed) was fed limited amounts of the control diet that matched the mean daily dietary intake of the PCM group. After 4 wk on the diets, alveolar macrophages from all three groups functioned similarly with respect to surface adherence, phagocytosis and killing of Listeria monocytogenes, release of hydrogen peroxide and superoxide anion, and production of IL-1 and TNF. In contrast, Listeria-stimulated alveolar macrophages from the PCM group exhibited a marked shift in arachidonic acid metabolism, with impaired production of leukotriene B4 and enhanced release of thromboxane B2 and PGE2. The membrane arachidonic acid content and the uptake of [3H]arachidonate by alveolar macrophages did not differ among the three groups. The shift toward the cyclooxygenase pathway was not seen after 2 wk of dietary restriction and was reversed if PCM animals were fed the control diet for 1 wk. Thus, PCM does not affect the antimicrobial activity or cytokine production of alveolar macrophages, but causes alterations in arachidonic acid metabolism that may interfere with the modulatory functions of alveolar macrophages.     

Effect of corticosterone and protein malnutrition on muscle protein breakdown in vivo in rats as measured by the urinary excretion of 3-methylhistidine

The role of corticosterone in regulating the rate of muscle protein breakdown was evaluated by measuring the urinary excretion of 3-methylhistidine (3-Mehis) during the administration of 0.0 (vehicle), 0.8 (physiological dose) and 10 (pharmacological dose) mg of the glucocorticoid/100 g body weight/day to adrenalectomized rats (AdX, AdX 0.8 and AdX 10 respectively). A fourth group of intact rats receiving only vehicle (In) was included as control. Rats were fed on either adequate protein and energy (Co) or low-protein (1-P) diets, for eight consecutive days. No differences were found between AdX and AdX 0.8 groups as compared to the In group in regard to body and liver weights. The AdX 10 group exhibited a significant reduction in body weight and a considerable increase in liver weight; these results were found in rats fed on the Co and 1-P diets, although rats on the 1-P diet showed a proportional decrease in those parameters as compared to the rats fed on the Co diet. Gastrocnemius, tibialis and E.D.L. muscle weights were significantly reduced in AdX 10 group, approximatley at the same extent in the two dietary groups. Soleus muscle weight increased in the AdX 10 group, at the same extent in the two dietary groups, as compared to the In group. Plasma corticosterone levels were significantly greater in the AdX 10 group in both dietary treatments, though restriction of protein in the diet induced a higher plasma hormone level than that of the Co group. Urea-N and creatinine outputs were significantly higher in the AdX 10 group. 3-Mehis excretion underwent an immediate and significant rise in the AdX 10 group, although rats fed on 1-P diet showed a more persistent rise than those fed on the Co diet. No differences were found among the other groups. It is concluded that high plasma corticosterone levels can accelerate muscle protein breakdown and that this action is not seriously affected by the protein content of the diet.     

Effect of dietary nucleotides and orotate on the blood levels of prostacyclin (PGI2) and thromboxane (TXA2) in the weanling rat

Dietary nucleotides affect the maintenance of immune responses, tissue repair and polyunsaturated fatty acid metabolism. Orotate, a pyrimidine nucleotide precursor, induces fatty livers by impairing VLDL hepatic secretion. The aim of this study was to evaluate the changes in the blood levels of fatty acids and prostacyclin (PGI2) and thromboxane (TXA2) in the weanling rat caused by the dietary intake of nucleotides and orotate. Three groups of rats at weaning were fed a control diet, an orotate supplemented diet (O-50) and a nucleotide supplemented diet (N-50) during 4 weeks, respectively. Absolute values of plasma polyunsaturated fatty acids greater than 18 carbon atoms of the n-6 and n-3 series were increased in the N-50 group and decreased in O-50 with regard to the control. However, the relative fatty acid composition of plasma lipid fractions was mostly unaffected. Plasma 6-keto-PGF1 alpha showed a trend to be increased in N-50 and serum TXB2 was significantly increased in that group. Both eicosanoids were unchanged by dietary orotate intake. These results may be explained because of the increased plasma 20:4n-6 found in rats fed a supplemented nucleotide diet. Thus, nucleotides present in foods appear to modulate PUFA conversion and eicosanoids synthesis in early life.     

Hormonal imbalance and disturbances in carbohydrate metabolism associated with chronic feeding of high sucrose low magnesium diet in weanling male wistar rats

This study was designed to determine chronic effect of high sucrose low magnesium (HSLM) diet in weanling rats on plasma thyroid profile, catecholamines and activities of key hepatic glycolytic, and gluconeogenic enzymes. Compared to control diet fed group, significantly elevated levels of plasma triiodothyronine, tetraiodothyronine, catecholamines (epinephrine, norepinephrine, and dopamine) and activity of hepatic glycolytic (hexokinase and glucokinase), and gluconeogenic (glucose-6-phosphatase) enzymes were observed in high sucrose and low magnesium fed groups. However, HSLM diet had an additive effect on all these three parameters. The study thus, assumes significance as it shows that hormonal imbalance and disorders in carbohydrate metabolism at an early stage of development can be due to dietary modification or due to deficiency of key element magnesium.     

Blood pressure of rats as affected by diet and concentration of NaCl in drinking water.

In the first experiment, weanling rats were fed a grain ration or one of three semipurified diets high in fat, sucrose, or cornstarch. Rats in each dietary group were divided into two subgroups, one of which drank distilled deionized water whereas the other group drank 2% NaCl solution. Blood pressure and sodium intake were individually measured for each rat at weekly intervals for a 10-week period. Rats receiving the salt solution had higher mean blood pressures (127-178 mmHg) than rats offered distilled water (108-127 mmHg). When drinking solutions were the same, more severe rises in blood pressure occurred in rats fed the semipurified diets than in those rats fed grain. In a second experiment, rats were fed one of the four diets used in the first experiment; however, they received a 1% NaCl drinking solution for 9 weeks followed by a 1.5% NaCl solution for an additional 9 weeks. At Week 18, pressures among these groups of rats ranged from 136-140 mmHg, regardless of diet.     

Effect of protein deficiency on macroelement and trace element levels of weanling rats' small intestine and liver tissues

Protein energy malnutrition has become a major health issue in developing countries. In the present study, the effect of protein deficiency on the small intestine and liver tissue content of macroelements and trace elements was investigated in weanling rats. Forty-five male weanling Wistar albino rats were divided into three groups. The control group (C) was fed a standard diet containing 25% casein, whereas the two experimental groups E1 and E2 consumed 12% and 3% casein, respectively, over a period of 45 d. The tissue samples were analyzed for zinc, copper, iron, manganese, calcium, and magnesium by atomic absorption spectroscopy. The protein-deficient groups showed increased levels of iron in both tissues and decreased manganese in small intestine tissue from the E1 group. No other differences were found for the other elements. These results suggest that protein deficiency might cause iron accumulation in the liver and intestine and decreases of manganese in the small intestine.     

Influence of diet on the development of nephrocalcinosis in the rat

A histological examination of the effect of a purified diet containing 20% alpha protein (an alkali-treated soyprotein) on the development of nephrocalcinosis induced by intraperitoneal injections of 0.5 neutral (pH 7.4) sodium phosphate was carried out in female weanling rats. Animals that were fed a standard commercial laboratory diet and given daily injections of phosphate for six or ten days developed a form of nephrocalcinosis that consisted mainly of intraluminal (intratubular) calcification at the junction of the outer and inner stripes of the outer medulla and in the inner stripe of the outer medulla. By contrast, rats that were fed the alpha protein diet and given injections of phosphate for six or ten days developed a form of nephrocalcinosis that was characterized primarily by a type of tubular basement membrane calcification at the junction of the inner stripe of the outer medulla and the inner medulla. The differences in nephrocalcinosis between the two dietary groups and the fact that an alpha protein diet by itself can cause renal calcification, leads to the suggestion that some component(s) or factor(s) in the alpha protein diet strongly influence(s) the development of nephrocalcinosis induced by injected neutral sodium phosphate.     

Dietary vitamin E deficiency increases anxiety-like behavior in juvenile and adult rats

Vitamin E deficiency from birth or infancy has recently been found to increase anxiety-like behavior in rodents. The present study was undertaken to elucidate the effect of dietary vitamin E deficiency on anxiety in adult rats in comparison with juvenile rats. Male Wistar rats, 3 or 10 weeks old, were divided into two groups and fed a control or vitamin E-deficient diet for 4 weeks. The results of behavioral analysis revealed that vitamin E-deficiency increased anxiety in both juvenile and adult rats. Plasma, liver, and brain α-tocopherol concentrations decreased significantly due to vitamin E deficiency in both age groups. Plasma corticosterone concentrations were higher in the vitamin E-deficient rats in response to the stress of a behavioral test. Based on these results, we conclude that dietary vitamin-E deficiency induces anxiety in adult rats as well as juvenile rats. This might be due to an elevated plasma corticosterone concentration.     

Dietary Vitamin E Deficiency Increases Anxiety-Like Behavior in Juvenile and Adult Rats

Vitamin E deficiency from birth or infancy has recently been found to increase anxiety-like behavior in rodents. The present study was undertaken to elucidate the effect of dietary vitamin E deficiency on anxiety in adult rats in comparison with juvenile rats. Male Wistar rats, 3 or 10 weeks old, were divided into two groups and fed a control or vitamin E-deficient diet for 4 weeks. The results of behavioral analysis revealed that vitamin E-deficiency increased anxiety in both juvenile and adult rats. Plasma, liver, and brain α-tocopherol concentrations decreased significantly due to vitamin E deficiency in both age groups. Plasma corticosterone concentrations were higher in the vitamin E-deficient rats in response to the stress of a behavioral test. Based on these results, we conclude that dietary vitamin-E deficiency induces anxiety in adult rats as well as juvenile rats. This might be due to an elevated plasma corticosterone concentration.     

Development of copper deficiency in rats fed fructose or starch: weekly measurements of copper indices in blood

Copper deficiency was induced in weanling rats fed diets whose sole source of carbohydrates was starch or fructose for 7 weeks. Conventional parameters of copper status, plasma copper concentrations, ceruloplasmin activity, and erythrocyte superoxide dismutase (SOD) activity were longitudinally monitored weekly to follow the development of the deficiency and to correlate these indices with the degree of severity of the deficiency. Although 30% of the rats fed a copper-deficient fructose diet died and no deaths occurred in rats fed the copper-deficient starch diet, plasma copper, ceruloplasmin, and SOD activities were reduced to a similar extent in all rats fed copper-deficient diets regardless of the type of dietary carbohydrate. Thus, none of the indices used accurately reflected the greater degree of deficiency or mortality in rats fed the fructose diet deficient in copper. The results of the present study underscore the need for more sensitive tests or alternative parameters to assess copper status in living animals.