Collagen-induced arthritis in rhesus monkeys: evaluation of markers for inflammation and joint degradation.

The objective of this study was to analyse parameters in rhesus monkey collagen-induced arthritis (CIA) with which the inflammation and destruction of the joints can be described in quantitative terms. CIA was induced in genetically susceptible and resistant monkeys, which can be distinguished on the basis of the dominant resistance marker Mamu-A26. The disease course was monitored daily using a semiquantitative scoring system. Plasma samples were collected once or twice weekly and analysed for C-reactive protein (CRP). Urines were collected overnight once a week and analysed for excretion rates of the collagen cross-links hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP). The results show that periods of active CIA are characterized by substantial weight loss and increased plasma CRP levels, followed shortly thereafter by increased excretion rates of the collagen cross-links HP and LP. Remission of the disease can be recognized by a decline in plasma CRP levels and especially an increase in body weight. The highest CRP levels were found in the most severely arthritic monkeys, indicating a possible relationship of the absolute plasma CRP levels to the severity of inflammation. During periods of active arthritis, increased excretion rates of collagen cross-links HP and LP in the urine were found. In particular, the major collagen cross-link in articular cartilage, HP, showed a strong increase (9- to 15-fold). The excretion rates of LP, which is considered as a bone-specific degradation marker, only increased 4- to 6-fold, thus indicating predominant destruction of cartilage and less of bone. In conclusion, the severity of CIA can be monitored in a quantitative manner using plasma CRP levels, urinary excretion rates of HP and LP, and body weights, superimposed on semiquantitative clinical scores. The parameters also facilitate a more objective assessment of the effect of anti-arthritic drugs in the model than with the clinical scores alone.

The role of interleukin-6 in the induction of hypercalcemia in renal cell carcinoma transplanted into nude mice.

Hypercalcemia is a well known complication of renal cell carcinoma (RCC). As RCCs can produce IL-6, and IL-6 may stimulate bone resorption and cause mild hypercalcemia, we examined whether IL-6 is involved in renal cancer-associated hypercalcemia in vivo. Three human renal cell carcinoma tumor lines (RC-8, RC-9, and NC-65) growing in nude mice were studied. Tumors were implanted s.c., and parameters of bone metabolism and serum human IL-6 levels were determined in relation to tumor volume (TV). All three tumor lines secreted human IL-6, although in different quantities. The maximum level of IL-6 in RC-8 was 434 pg/ml (TV, 200 mm3), that in RC-9 was 81 pg/ml (TV, 1800 mm3), and that in NC-65 was 2368 pg/ml (TV, 1800 mm3). Hypercalcemia developed in RC-8 and RC-9 tumor-bearing animals, but not in NC-65-bearing animals. The hypercalcemia in both RC-8 and RC-9 tumor lines was associated with elevated levels of PTH-related peptide (PTHrP) and loss of trabecular bone volume. Serum calcium and phosphate concentrations showed an almost linear relationship with plasma PTHrP independently of the tumor line and serum IL-6 levels. No hypercalcemia occurred in the NC-65 animals, which had the highest levels of IL-6, but no detectable plasma PTHrP and PTHrP messenger RNA expression in the tumor. Administration of neutralizing antibodies to IL-6 to RC-8 animals normalized serum calcium concentrations and PTHrP values and induced a significant inhibition of tumor growth. No such effect on tumor growth of anti-IL-6 was seen in the other two tumor lines. The normalization of serum calcium in RC-8 mice is most likely attributed to the growth-inhibiting effect of anti-IL-6 on RC-8 tumor. We conclude that IL-6 secreted by RCC does not contribute directly to hypercalcemia, but may enhance hypercalcemia by stimulating the tumor growth of a subpopulation of PTHrP-secreting carcinomas.

[The importance of mineral supplementation for dentition and jaw]. / Het belang van mineraalsuppletie voor gebit en kaak.

In this article the importance of mineral suppletion for teeth and jawbone is explained in a step-wise manner. The steps discussed are the pre-eruptive phase of teeth and the growth, adult and ageing phase of the skeleton. Mineral suppletion should be seen as a measure to prevent dental caries (fluoride) and to ensure a maximal resistance against the spontaneous bone loss during ageing. Therefore, it seems not suitable for therapy of osteoporosis. If an enhanced bone loss occurs as a result of decreased production of sex hormones or immobility, mineral suppletion will probably have a minor effect. An optimal mineral intake will ensure a maximal peak bone mass built up during the growth phase of bone, a maintenance of this peak bone mass as much as possible during the phase of neutral bone balance and a minimal bone loss during the ageing period with negative bone balance.

Evidence for a diminished maturation of preosteoblasts into osteoblasts during aging in rats: an ultrastructural analysis.

Bone is subject to continuous remodeling throughout life. The age-related loss of (trabecular) bone, leading to senile osteopenia, is mainly due to impaired bone formation. Osteoblasts (OB) and osteoclasts (OC) have been identified as playing a crucial role in the process of bone turnover, but the contribution made by their precursors is not well documented. We analyzed the cells of the osteoblast and osteoclast cell lineage along the trabecular bone of tibiae and the stromal cells in the marrow of aging BN/Bi Rij rats using electron microscopy. It appeared possible to distinguish preosteoblasts (pre-OB), OB, preosteoclasts (pre-OC), OC, and inactive bone-lining cells. Periods of increase, the maximal peak, and the decrease in trabecular bone volume were defined by means of morphometric measurements of trabecular bone volume. We found a decrease of more than 10-fold in the number of OB with age, but the numbers of pre-OB, pre-OC, and OC expressed per unit bone length, although variable, were age independent. The relative bone resorption and formation surface, expressed as a percentage of the total bone surface, decreased 2- and 15-fold, respectively. In 2-year-old animals the total volume of stromal cells, part of which constitutes the stem cell compartment of the osteogenic lineage, was a quarter of that found in 1-month-old animals and a third of that found in 6-month-old animals. The loss of trabecular bone is concomitant with a sharp increase in the ratio of pre-OB/OB, the ratio of OC/OB, and in the ratio of resorption to formation surfaces. There was no relation between the ratio of pre-OC/OC with age. These data lead to the conclusion that the main factor causing bone loss with age is a diminished maturation of pre-OB into OB.

Caries experience of 15-year-old children in The Netherlands after discontinuation of water fluoridation.

In 1973 the fluoridation of drinking water in the Dutch town of Tiel was discontinued. In order to monitor the effect of this measure, the caries experience in 15-year-old children was investigated annually from 1979 to 1988, both in Tiel and in Culemborg. In the latter town the drinking water had never been fluoridated. The caries data of 15-year-old children examined between 1968 and 1969 in Tiel (children having used fluoridated water from birth) and Culemborg were used as historical controls. In Tiel the mean number of DMFS increased between 1968/69 and 1979/80 from 10.8 to 12.7 (+18%) and decreased to 9.6 (-26%) in the following years; in 1987/88 the mean DMFS was 11% lower than in 1968/69. In Culemborg the mean DMFS score decreased between 1968/69 and 1987/88 from 27.7 to 7.7 (-72%). In 1968/69 the mean DMFS score in Tiel was 61% lower and in 1987/88 17% higher than in Culemborg. The question as to whether water fluoridation would have had an additional effect if it had been continued (presuming the application of existing preventive measures) cannot be answered, as there are no remaining communities with fluoridated water in The Netherlands.

Relationships between birefringence and mineral content in artificial caries lesions of enamel.

The microradiographic mineral content and birefringence in water and Thoulet's solution were measured at selected points in sections of caries-like lesions. Birefringence was not related to mineral content in sound superficial enamel immersed in Thoulet's solution or in the lesion body immersed in water. For the surface layer of the lesion, birefringence in water could be used to obtain qualitative information about mineral content. For the lesion body, birefringence in Thoulet's solution was linearly related to mineral content and can be used to estimate the latter with a standard error of about 7 vol%. The intrinsic birefringence for the lesion body was estimated as -34.5 (+/- 2.3) x 10(-4); This value may also be valid for other parts of the lesion. From consideration of ionic sizes, it is argued that Thoulet's solution cannot necessarily penetrate all enamel pores accessible to water. It appeared that there is a progressive fall in the content of pores inaccessible to Thoulet's solution in the surface layer of the lesion as demineralisation increases.

[Cessation of fluoridation of drinking water; results of caries research in Tiel and Culemborg in the period of 1968-1988]. / Stopzetting van drinkwaterfluoridering. Resultaten van het cariësonderzoek in Tiel en Culemborg in de periode 1968-1988.

In December 1973 water fluoridation was discontinued in Tiel, the Netherlands. Six years later an epidemiological study was started, with the aim to investigate the consequences. In the period from 1979 to 1988 a yearly caries investigation was carried out in 15 year old children born and living in Tiel and children living in the control town of Culemborg, where the drinking water had never been fluoridated. The 15 year old children in 1979-1980 in Tiel had a 18% higher caries experience than the same age group in 1968-1969, to whom fluoridated water had been available from birth onward. In the same period a 28% decrease of the caries was found in Culemborg. In 1979-1980 the DMFS-index in Tiel was still 36% lower than in Culemborg. Between 1979-1980 and 1987-1988 the caries experience decreased in both towns.

Vitamin D, within its range of fluctuation in commercial rat diets, does not influence nephrocalcinogenesis in female rats.

Massive, toxic doses of vitamin D have been shown to cause nephrocalcinosis in rats, but the effect of this vitamin within its range of fluctuation in commercial rat diets was unknown. Therefore, in two experiments with young female rats, the effect on nephrocalcinosis of a moderately increased level of vitamin D in the diet was studied, that is 5000 IU/kg versus the recommended concentration of 1000 IU/kg. This was done using purified diets with 0.5% (w/w) calcium and 0.04% magnesium containing either 0.2 or 0.6% phosphorus (P). Rats fed the diets containing 0.6% P showed severe kidney calcification compared to those fed the 0.2%-P diets. The level of vitamin D in the 0.2 and 0.6%-P diets did not affect kidney calcification. Bone density was increased after feeding diets containing 5000 instead of 1000 IU of vitamin D/kg. This study suggests that, within 28 days, a moderate increase of the amount of vitamin D in the diet has no influence on the development of kidney calcification. This in turn suggests that the variation in nephrocalcinosis severity and incidence seen in practice in rats fed different commercial diets is unlikely to be related to the different vitamin D concentrations in these diets. However, in rats fed such diets bone metabolism may be influenced differently.

Electron microprobe and microradiographic studies of carious enamel lesions covered with dental calculus.

From a large number of posterior teeth with arrested natural carious lesions some 0.56 percent were found to have arrested approximal lesions with a superficial layer of calculus. The lesions were examined using quantitative microradiography for mineral content and by electron microprobe analysis for Ca, Na, Cl, Mg and P. The conditions in the plaque, which caused the formation of calculus, might also be expected to cause a remineralization of the lesions. However, the lesions examined were found to have a low mineral content in their body, and moreover, had a lower Na/Ca ratio and a higher Cl/Ca ratio than sound enamel; these are characteristics also found in carious lesions not covered with dental calculus. On the assumption that the lesions developed before any plaque calcification occurred, it is concluded that conditions which favour the calcification of plaque do not favour the remineralization of carious lesions in vivo. As only 0.5% of the carious lesions in extracted teeth were covered with calculus, it must be admitted that consecutive periods of lesion formation followed by calculus formation on the same tooth face are rare.

Microradiography and electron-microprobe analysis of some caries-like lesions of enamel prepared in vitro in human teeth.

Six lesions were made on the buccal surfaces of premolars. The volume percentage of mineral was determined as a function of depth by microradiography. Using the electron microprobe, the signals for Ca, Na, Mg, P and Cl were recorded as a function of depth both through the lesions and through the adjacent sound enamel. In the demineralized parts, there was a preferential loss of Na and a preferential retention of chloride. In the surface layers, the Na:Ca and Cl:Ca ratios were almost the same as in the adjacent sound enamel, indicating that the surface layers were not formed by gross dissolution of the original mineral followed by gross reprecipitation of another, less-soluble calcium phosphate, but remained probably because their microcrystals were protected by a thin layer of precipitated fluorapatite or fluoridated hydroxyapatite. The same had been found for surface layers in natural caries.