IBMS/ECTS2001 - ABSTRACTS: Late Posters

MANDIBULAR BONE LOSS IN AN ANIMAL MODEL OF MALE OSTEOPOROSIS (THE ORCHIDECTOMIZED RAT): A RADIOGRAPHIC AND DENSIOMETRIC STUDY

E. Lerouxel, H. Libouban, M.F. Moreau, E. Legrand, M.F. Baslé, M. Audran, D. Chappard*

In human, hypogonadism is associated with osteoporosis and can be appreciated by densitometry (DXA) on the vertebrae or long bones. However, there are some controversies about the relationships between bone loss in these sites and in the mandible. Osteoporosis has been suggested as a risk factor for implant failure, but data supporting such a hypothesis are limited. In the rat, orchidectomy (ORX) is associated with an accelerated bone turnover and increased bone resorption resulting in bone loss. We have studied the time effects of ORX on the alveolar bone of the mandible. 48 male Wistar rats (18-19 weeks old) were divided into 4 groups and studied during 2, 4, 8 and 16 weeks. In each group, 6 rats were ORX and 6 non-ORX were used as control. The mandible of each rat was dissected, hemisected at the symphysis and all soft tissue removed. The rat mandible has a complex structure with subparts composed of thin cortices and other containing both cortical and trabecular envelopes, especially in the molar region. Two types of numerized radiographs were obtained with a Faxitron machine for each hemimandible; a general global view and an enlarged view centred on the molar region. Bone loss was observed qualitatively at 16w. in ORX animals, as the texture of the alveolar bone appeared coarser. Quantitative modifications were confirmed by texture analysis. The mandible mineral content was measured with a Holologic QDR4500 on each left hemimandible. BMD and BMC measurements were poorly reproducible on the whole bone and no differences could be observed for BMC between ORX and controls, even at 16w. When BMS was considered in the molar region, a significant bone loss became evident in the ORX group at 16w (p<0.01). In a male rat, the reduction of sex hormones induced by ORX is also associated with a decrease in bone mass in the mandible.

Department of Psychiatry, Department of Medicine, University of Heidelberg, Germany

The potential association between schizophrenia and osteoporosis and/or osteopenia has only recently been recognized. Osteoporosis and/or decreased BMD has been attributed to polydipsia, to nicotine and alcohol abuse in schizophrenic patients, and to lack of UV exposure and/or vitamin D. Furthermore, osteopenia and/or osteoporosis have been attributed to drug-induced decreases in estrogen and testosterone levels, to hyperprolactinemia and to increased activity of several interleukins. As there is little empirical evidence from larger patient cohorts, the present study was carried out.

Sixty-nine premenopausal, regularly menstruating schizophrenic women (mean age 33.3, SD 6.2; range 18 to 45 years) consecutively admitted to the University's Psychiatric Unit, and 68 age- and sex-matched healthy controls were included in the study. Laboratory parameters were urinary pyridinoline (PYD) and deoxypyridinoline (DPD; by HPLC) as well as serum osteocalcin (OC; by LIA), parathyroid hormone (iPTH; by EIA), 25 hydroxyvitamin D (25OH-D; by RIA), insulin-like growth factor 1 (IGF-1; by EIA), estradiol (E2; by EIA) and testosterone (by EIA). In the patients, measurements of spine and hip BMD were performed by DEXA.

Compared to controls, a significant increase in urinary PYD and DPD, and in serum OC was observed in the schizophrenic women. In addition, iPTH was increased and 25OH-D was decreased in the patients, while no difference was found in serum IGF-1 levels. Furthermore, estradiol was decreased and testosterone was increased in schizophrenic women as compared to controls. However, in a subgroup of 57 schizophrenic women, BMD was within the normal range. There was no correlation between BMD and the duration of disease.

We conclude that premenopausal and regularly menstruating schizophrenic women have normal spine and hip BMD, despite significantly increased bone turnover. This may be due to the opposite effects of the different parameters influencing bone metabolism, especially of the gonadal hormones, and due to continued coupling.

EFFECT OF ADVANCED GLYCOSYLATION END-PRODUCTS ON THE PROLIFERATION AND DIFFERENTIATION OF RAT'S OSTEOBLASTIC CELL

Dept of Biochemistry, HeBei Medical University, Shijiazhuang 050017, P. R. China

Advanced glycosylation end products (AGEs) had played a lot of pathophysiological roles in vivo. Clinical studies showed that the modification of ß2-microglobulin with AGEs might play a pathophysiological role in osteoporosis around amyloid deposits, associated with dialysis-related amyloidosis. The aim of this study was to explore effects of AGEs on the proliferation and differentiation of osteoblastic cell in order to reveal the pathogenesis of the senile osteoporosis.

We chose to prepare AGE-protein by means of the incubation of BSA and glucose. To assess time-dependent and dose-dependent pattern of AGE-BSA effect on rat’s osteoblastic cell proliferation and differentiation, cells were incubated for different periods of time after exposure to different doses of BSA or AGE-BSA.

After 48h incubation with 50~250µg/ml AGE-BSA, there were no significant differences between the effect of BSA and AGE-BSA on cell proliferation. While 500µg/ml AGE-BSA induced the inhibition of cell proliferation. After 72h incubation with 50-500µg/ml AGE-BSA or BSA, no significantly differences were found between the effect of AGE-BSA and BSA on cell proliferation. But after 96h incubation, 50µg/ml AGE-BSA significantly stimulated cell proliferation, while 500µg/ml AGE-BSA significantly induced the inhibition of cell proliferation. When alkaline phosphatase (ALP) activity was studied, cells exposed to 50µg/ml AGE-BSA increased their ALP activity for 24h incubation. After 72h incubation in the presence of 50-250µg/ml AGE-BSA or BSA, AGE-BSA increased the ALP activity. But there was no significantly difference between the effect of AGE-BSA and BSA on cellar ALP activity at dose of 500µg/ml AGE-BSA.

These results had shown that low levels of AGE-BSA increased rat’s cell proliferation, high levels inhibited the cell proliferation; low contents of AGE-BSA stimulated cell differentiation as early as 24h after exposure to AGE-BSA, but high contents of AGE-BSA induced the cell differentiation as lately as 72h, which resulted in the decrease in the amounts and function of osteoblast relatively. Several studies indicated that AGEs could combine with their receptor on some cell membrane to induces these cells to synthesis and release beta interleukin-1 (IL-ß), alpha tumor necrosis factor (TNF-a), interleukin-6 (IL-6), These cytokines stimulated the osteoclast precursors to get into mature osteoclast and increased their functions, which resulted in the excess of bone resorption relatively.

The pathogenesis of senile osteoporosis may be that excess of AGEs with age induced the inhibition of osteoblastic cell proliferation and the delay of differentiation, while the amounts and function of osteoclast increased relatively, which caused that bone loss is more than bone formation and so the osteoporosis occurred.

THE PREVENTION AND TREATMENT OF TONIFYING ESSENCE OF THE KIDNEY RECIPE FOR THE RAT OSTEOPOROSIS

Institution of Traditional Chinese and Western Medicine Basic Theory, HeBei Medical University, Shijiazhuang, P. R. China

With the coming of aging society, osteoporosis will become one of great problems endangering senile people health. The current therapeutic agents for osteoporosis were bone absorption inhibitor. Hormone replacement therapy was used for postmenopausal osteoporosis.

In the Traditional Chinese Medicine theory, "the kidney is the foundation of the native constitution". "The condition of the kidney determines the condition of growth and development". The kidney stores the essence of life either congenital or acquired. The bone marrow is full if the kidney is good condition. The bone could not obtain fully nourishment if the essence of kidney is deficiency. Basing on these theories, the rats received with Dexamethasone or the ovariectomized rats or senile were given tonifying essence of the kidney recipe (TKR) to protect rats from bone mass loss. The bone mineral density and bone metabolism-associated biochemical markers were determined. Bone histomorphometric indexes were surveyed. Results: (1) Tonifying essence of the kidney recipe (TKR) can protect the rats received by Dexamethasone from bone mass loss. TKR can significantly increase the levels of serum osteocalcin, decrease the excretion of Ca in urine. Southern blotting analysis showed that the TKR can promote the expression of the CaBp-D9K gene so that the increase in CaBp production was preceded by the expression of CaBp-D9K mRNA, which promoted the absorption of Ca in the intestine.(2) TKR protected the ovariectomized rats from bone loss, increase the production of type I collagen in bone tissue and the expression of the vitamin D receptor(VDR) mRNA in the cells in the intestine mucosa and suppressed MMp-9 activity in bone tissue. TKR can significantly increase the levels of serum estrogen in the ovariectomized rats. Immunohistochemical method, Western blotting analysis and reverse transcription -polymerase chain reaction analysis showed that TKR promoted the expression of estrogen receptor gene in osteoblast by which bone metabolism was regulated by estrogen. (3) Results of bone histomorphometric study: trabecullar bone volume % (TBV%), trabecullar bone formation surface %(TFS%) and trabecullar bone surface %(TRS%) from senile rats' tibias were lower than that from young rats' tibias. These results suggested that bone metabolism of senile osteoporosis belonged to low turnover. TBV%, TFS% and TRS% from tibias of senile rats administered with TKR were significantly higher than that from senile rats' tibias. TKR decrease the level of urine Ca/Cr in senile rats.

The above results suggested that the TKR modulated the bone formation and bone absorption so that the couple of bone formation and absorption occurred to prevent bone mass loss in many ways.

EXPERIMENTAL RESEARCH AND CLINIC OBSERVATION OF ON THE EFFECT OF DUZHONG LEAVES EXTRACTS IN THE TREATMENT OF PRIMARY OSTEOPOROSIS

Department of Biological Engineering, Jiang Xi Normal University, Nanchang, 330027, P. R. China

Department of Orthopedics, Jiang Xi College of Traditional Chinese Medicine, Nanchang, 330006, P. R. China

Primary osteoporosis (POP) is one of normally and frequently occurred diseases which harms middle and old-aged-people’s health. (Members of our study group have engaged in the study of preventing and treating POP with Chinese medicine for many years). In order to seek safe, ideal and effective medicine and therapy, precious tonifying and nourishing Chinese medicine - Duzhong leaves (Folium Eucommiae) were used as raw materials, and were abstracted and separated by column chromatography into five parts - A, B, C, D, E. These extractive parts were studied in influencing metabolism and proliferation of osteoblast cultured in vitro. The cranial bones of new born SD rats were used for primary culture, and the secondary subculture cells were used for experiment. Estraoliol were used as drug contrast group, a blank contrast group was set up. Testing indexes: (1)cell proliferation calculated with MTT testing method;(2)ALP activity tested with colorimetric analysis. The study showed that part B and part D were effective contrasted with blank group in activating secreting ALP and in promoting cell proliferation P<0.001.

According to Chinese medical theory of syndrome differentiation, POP patients are classified into four types. The old recipe - Qin-Er Pill decreasing and increasing were used in treatment of 42 POP patients according to syndrome differentiation, in which the extractive parts B and C were used as ‘monarch’ drug. The period of treatment was 3 months. Objective detecting indexes including bone mineral density (BMD), biochemical markers of bone metabolism and integrals of kidney-deficiency were used. a-D3 was also used as contrast group (30 cases). The results of two groups were obviously different compared with that of pretreatment, the intetrals of kidney-deficiency varied even obviously, treatment group P<0.01; control group (P<0.05). It showed that the effect of the extracts of Duzhong leaves - part B and part C in treatment POP is contented, it proved that the two parts have estrogen-like effect.

³Departamento de Enfermería. Escuela Universitaria Ciencias de la Salud, Universidad de Granada

Osteoblasts express in their surface CD13, antigen with aminopeptidase activity. Such an ectoenzyme is also present on the surface of many other cell types and seems to participate in the processing (activation or inactivation) of peptides released to the extracellular medium by different cell types, including the same osteoblasts.

We have studied the effects of several non-esteroidal antiinflammatory drugs (NSAIDs), such as piroxicam, indomethacin, aspirin and naproxen on aminopeptidase activities in cultures of human osteoblasts from samples of normal bone, obtained during mandibular osteotomy. Previously, the cells were characterised on the basis of their morphological, biochemical (alkaline phosphatase activity and osteocalcin synthesis) and antigenic phenotype. A fluorimetric technique was used to examine the enzymatic activity on different artificial substrates. We detected an inhibitory effect of NSAIDs on aminopeptidase activities of osteoblasts in 7 of the 15 substrates analysed. Piroxicam and indomethacin inhibited enzymatic activities with a IC50 in the micromolar range. The effect was dose dependent, showing a complete inhibition at the higher doses of the NSAID. Naproxen showed less inhibitory capacity than the two previously mentioned NSAIDs. Aspirin, on the contrary, was without effect.

Our results suggest that some NSAIDs may have a modulator effect on the possible role that aminopeptidases play in the turnover of peptides and proteins in the processes of bone remodelling.

Hypoxia is well known to act as a general stimulator of the recruitment and/or activation of cells derived from marrow precursors but its effects on osteoclasts (OC) have not been studied. We investigated the effects of oxygen tension (PO2) on bone resorption using two systems. Firstly, mouse marrow cells, free of stromal cells, were cultured 7 days on dentine discs in medium supplemented with 10 ng/ml RANKL, 30 ng/ml M-CSF and 10% serum in atmospheres containing 20%, 12.5% or 5% O2 (all with 5% CO2, balance N2). Culture medium was acidified to pH 7.0 for the final 2 days to activate osteoclastic resorption; PO2, PCO2 and pH were monitored by blood gas analyser. The 5% O2 environment resulted in a 4-fold stimulation of TRAP-positive OC formation, compared with 20% O2. However, OC formed in 20% O2 were generally small (1-2 nuclei), whereas those generated in 5% O2 were large, with multiple nuclei, resulting in striking increases in resorption pit formation (>15-fold). Secondly, ½ calvaria of 5 day old mice were incubated for 3 days in 2% or 20% O2 atmospheres on steel grids and Ca2+ release into culture medium was measured colorimetrically. OC-mediated Ca2+ release was >5-fold greater in the 2% O2 environment, equivalent to the maximum stimulation resulting from prostaglandin E2. Increased OC resorption in bones exposed to 2% O2 was evident in TRAP-stained whole mount preparations. Hypoxia-stimulated resorption was completely blocked by 0.1 microM indomethacin, implying prostaglandin mediation, and was strongly inhibited by an IL-1 receptor antagonist protein. In calvarial but not marrow cultures, hypoxia was associated with significant culture medium acidification, consistent with increased anaerobic metabolism. In atmospheric air, arterial and venous blood, PO2 is 160, ~95 and ~40 mmHg, respectively (20%, 12% & 5%). Interstitially, PO2 is <40 mmHg, whereas in hypoxic environments such as the poorly vascularised yellow fatty marrow associated with ageing, or in inflamed tissue, tumours and fracture sites, PO2 may be <10-20 mmHg. These pathophysiological states are associated with increased bone resorption and also local acidosis, which is now recognised as the key requirement for OC activation. Our experiments reveal a new control mechanism for bone resorption of major importance. The stimulation of OC function in hypoxia may be mediated via increased production of angiogenic cytokines such as TNFalpha, IL-1 and VEGF; the role of RANK/RANKL is presently under investigation.

EARLY TREATMENT WITH INTRAVENOUS PAMIDRONATE NORMALIZES VERTEBRAL BONE MASS IN A 20-MONTH OLD CHILD WITH OSTEOGENESIS IMPERFECTA (OI) TYPE 1

Sección Osteopatías Médicas, Hospital de Clinicas, Universidad de Buenos Aires, Argentina

Osteogenesis imperfecta is a heritable disorder with increased bone fragility and pathological fractures. Medical treatment has long been largely ineffective, but it has been recently demonstrated that patients may benefit with pamidronate treatment. A patient with OI type 1 that began intravenous pamidronate at 7 months old and his evolution during one year treatment is reported. CASE REPORT: A boy who was seen for the first time at 7 months of age. Family history: his mother and a grandmother with OI. Previous fractures: vertebrae, femur, humerus and ribs. Positive data on physical examination: blue sclerae, joint hypermobility, weight (5th percentile) and height (20th percentile)..X rays: Spine: generalized osteopenia with two fractured vertebrae. Bone mineral density (BMD) (DEXA): L1-L4 (g/ cm2) : 0.154 (Z: - 4). Laboratory: sCa : 10 mg/dl, sP: 5.6 mg/dl. AP: 490 IU/l, uCa/uCreat: 0.10. Intravenous pamidronate was administered in cycles of three days (beginning with a dose of 0.5 mg/kg/day, increased to 1 mg/ kg /day), given every 4 months. The patient had an acute phase reaction (short-term fever) and mild hypocalcemia only after the first infusion. Results of treatment: his growth was adequate and he has been a very active boy who did not have new fractures. There was a significant increment of 180% the lumbar spine BMD during the year of treatment, reaching normal values of BMD for the age: 0.432 g/cm2 (Z: +0.19) (Figure 1). Radiological examination showed increment of vertebral density and remodeling of the affected vertebral bodies.

CONCLUSION: Early intravenous pamidronate treatment is suggested in OI (inclusive in mild forms) because it was highly effective: no new fractures, marked remodeling of vertebrae, and acquisition of normal lumbar bone density in a high active boy with OI type 1.

Sección Osteopatías Médicas y División Traumatología y Ortopedia, Hospital de Clínicas. Buenos Aires, Argentina

There is scant of published data about bone mass measurements in men with appendicular fractures. The aim of this study was to evaluate bone mineral density (BMD) in a group of men that sustained one appendicular fracture. Twenty-one men that had suffered peripheral fractures (mean ± 1SD: 63 ± 8 years; range: 51-79 years of age) were admitted into the Hospital for treatment of their fractures (tibia:7;tibia and fibula: 3; humerus: 6; wrist: 3; pelvis and clavicle: one for each type of fracture). BMD of the spine and hip was measured by DEXA (Hologic QDR 1000) in the fractured men and 17 age-matched controls. The mean interval between fractures and the study was 19 ± 5 days. Patients with secondary causes of osteoporosis were excluded for the study.

Men that sustained appendicular fractures had statistically significant lower BMD over spine and trochanter. Z-score was statistically diminished over these regions but the difference did not reach significance over the femoral neck. These results indicate that trabecular bone is mainly affected at earlier age in this group of men. Conclusions: Measurement of lumbar spine could be useful in men that had suffer recently a peripheral fracture to evaluate bone loss and to decide therapeutical approaches to avoid the occurrence of further fractures in the future.

	Appendicular fractures (n= 21)	Controls (n =17)	%D	p
Age (Years)	62.8 ± 7.8	65.6 ± 4. 1		n.s.
Weight (Kg)	71.2 ± 10.5	74.1 ± 9.3		n.s.
Height (m)	1.68 ± 0.04	1.69 ± 3.7		n.s.
BMD (g/cm²)
L2-L4	0.820 ± 0.111	1.006 ± 0.082	-18	<0.001
Z-score	-1.8 ± 1.0	0.5 ± 0.9		<0.001
Femoral neck	0.777 ± 0.096	0.837 ± 0.088	-7	n.s.
Z-score	-0.2 ± 0.9	0.7 ± 0.7		n.s.
Trochanter	0.626 ± 0.090	0.724 ± 0.084		.05
Z-score	-0.8 ± 0.9	0.3 ± 0.6		<0.05
Total hip	0.836 ± 0.125	0.943 ± 0.097	-11	0.05
Z-score	-0.8 ± 1.0	0.3 ± 0.7		<0.05

BODY COMPOSITION AND TOTAL SKELETON BONE MINERAL DENSITY IN WOMEN WITH HIP FRACTURE

Sección Osteopatias Medicas, Hospital de Clínicas José de San Martín, Universidad Nacional Buenos Aires, Argentina

Different studies have reported that patients with osteoporotic vertebral fractures have lower body weight and fat mass than age-matched controls, however few studies have examined the body composition of these patients. The objective of this study was to evaluate the body composition and bone mineral density (BMD) of total skeleton by X-ray densitometry(DEXA) in a group of women with hip fracture compared to controls. We studied 42 women with hip fractures and 25 control women of similar age. Body composition and BMD of total skeleton and its sub-regions were obtained by DEXA(Lunar DPX-L) in patients and controls. The mean interval between fractures and the densitometric measurement was 17±9 days. The results were as shown below.

In hip fracture women lean mass and percentage of fat was significantly lower than controls and fat mass tended to be lower although did not reach statistically significance. Hip fracture patients had lower BMD skeleton and its subregions except arms. When patients with hip fracture were divided according to the type of fracture: cervical(n:23) or trochanteric(n:19), patient with trochanteric fractures were older and had significantly lower weight, body mass index, BMD in all regions measured and fat mass compared to patients with cervical fractures.

Body composition measurements by DEXA confirm that woman with hip fracture have significantly lower fat and lean mass. BMD was low in all skeletal areas. Our data showed that patients with trochanteric hip fractures were more severely affected according to all parameters studied than women with cervical hip fractures.

Sección Osteopatías Médicas, Hospital de Clínicas, Universidad de Buenos Aires, Argentina

Osteosclerotic diseases are infrequent pathologies of congenital or familiar and they are associated with sclerosis and abnormal bone turnover. A man with axial osteosclerosis was seen for the first time at 30 years old. Previous history: the patient has a previous diagnosis of osteopenia at 17 years of age, and he was treated with calcium. Three years ago started with pain in the thoracic and lumbar spine. The biochemical tests showed an increment of serum alkaline phosphatase and hydroxiprolyne excretion. The x-rays showed a uniform and symmetric increment of the radiographic density in pelvis, without alteration of the form and size. At that time the diagnosis of Paget´s disease was made and the patient was given oral pamidronate plus calcium for a period of 30 days.
Four months ago the patient was seen at the Sección Osteopatías Médicas of the University Hospital to reviewed the diagnosis. The patient complain of mild pain in the thoracic and lumbar spine.The patient was evaluated and the results are the following: laboratory: increment in one alkaline phosphatase (110UI/L, NV: 31-95), and urine crosslaps (743 µg/mmol creat, NV in men up to 400), tendency of low serum phosphate (2.9 and 3.0 mg%, VN 2.6-4.4). All the other tests were normal. The bone scan was normal. The densitometry showed and increment of BMD at L2-L4 1.725 g/cm² (+3.9 SD for age and sex controls), femoral neck 1.053 g/cm² (-0.1 SD) and total skeleton 1.332 g/cm² (+1.4SD). The trunk was the subarea of the total skeleton with the greater increment (+4.0 SD).

The x-rays showed uniform and symmetric sclerosis in the pelvis. The spine have horizontal lines with high density from the thoracic to the lumbar spine. With the provisional diagnosis of Axial Osteosclerosis a bone biopsy was indicated but the patient has refused so far to follow the recommendation. He will be treated with calcitriol in necessary dose according to tolerance and bone markers will be followed at frequent intervals.

Paget´s bone disease is frequent in our population while Axial Osteosclerosis is extremely rare. However it is seen to be a bone disease to consider in the differential diagnosis of Paget´s bone disease.

Sección Osteopatías Médicas and Servicio de Nefrología, Hospital de Clínicas, Universidad de Buenos Aires, Argentina

The aim of the present study was to evaluate the usefulness of serum CTX as a bone resorption marker in patients on hemodialysis. We study cross-sectionaly 60 control subjects and 50 patients on hemodialysis. Thirty four of these patients were studied at one year follow-up. Serum determination included: calcium, phosphorus, bone alkaline phosphatase(bAP), iPTH and sCTX. Results of the baseline study are shown in Table 1.
Serum CTX correlated significantly with the iPTH(r=0.74, p<0.001) and bAP(r=0.65, p<0.001) in the cross-sectionally study of patients on hemodialysis.

Longitudinal study: 34 hemodialized patients were evaluated longitudinally at one year (30 were still being hemodialized, 2 underwent parathyroidectomy and 4 had a renal tranplant). The results are expressed as a percentage of change compared with the previous year. The patients were divided into three groups according to the annual iPTH variations. Results are shown in Table 2.

A positive correlation between DsCTX and Di-PTH(r=0.84, p<0.001) was observed, this was higher than that between DbAP and DiPTH(r=0.53, p<0.001). The decrease in iPTH levels due to parathyroidectomy or renal transplant determined a similar decrease in sCTX levels.

Conclusion: sCTX is a more sensitive marker than bAP with respect to changes in iPTH and presumably of bone resorption in this disease.

RALOXIFEN (RLX): BENEFICIAL EFFECTS ON BONE MASS IN POSTMENOPAUSAL WOMEN - 2 YEARS FOLLOW-UP

J. J. Scali*, G. Castelli, S. Visentini, J. Salomón, P. Bárcena, E. Morales, G. Lancioni, R. Bolton, D. Sevilla

Unidad de Reumatología, Hospital Durand, Sede Coordinación Comisión Asesora de Reumatología, Secretaría de Salud, GCBA, Buenos Aires, Argentina

A new selective estrogen receptor modulator(SERM), RLX Clorhidrate was used to treat 30 postmenopausal women, between 40 to 75 years old, fulfilling the WHO Criteria for osteoporosis, during 24 months, in an open controlled trial, to determine the effectiveness to reduce risk of new vertebral and no vertebral fractures, evaluating efficacy to increase bone mass and also recording side effects. We exclude women suffering from other bone diseases, breast or uterine cancer in relatives or under calcitonin, biphosphonates and fluor treatment at least 6 months before starting the trial. They receive: Group RLX: 60mg/day with calcium supplementation (1.000mg/day) and colecalcipherol (400U/day) and Group Control: only Calcium supplementation and colecalcipherol (same dose). Patients were studied with hip and spine X-rays, looking for fractures to randomize in: a)-10 pts with previous fractures and b)-20 pts without previous fractures); Breast X-Rays, Bone densitometry (lumbar spine and hip) at baseline and when finishing the trial; general laboratory and bone markers (Alkaline phosphatase, osteocalcin, C-telopeptide and urinary pyridinolines.

RESULTS: Treatment compliance was acceptable (75% completed the trial). Women on RLX show increasing bone mass (2% in femoral neck and 2.4% in vertebral bodies)when comparing with controls. One patient abandoned treatment because of multiple fractures with excessive mineral density loss. One patient stopped the treatment because of RLX intolerance, 2 pts presented venous tromboembolic events (trombophlebitis), 3 complaints of legs cramps and 5pts presented head flushing, 2 edema in lower extremities, 6 pts presented arterial hypertension, 4 display high seric levels of colesterol and one case presented haematuria. We didn`t find any hematological, renal or liver adverse effects.

CONCLUSIONS: RLX was usefull to treat osteoporotic postmenopausal women, because its beneficial effects to increase bone mineral density in spine and femoral neck, with a good compliance and tolerance.

Department of Medical Physics and Medical Engineering, The University of Edinburgh, Western General Hospital, Edinburgh, EH4 2XU, UK

Due to the role of bone mineral contribution to the skeletal integrity, quantitative methods of its evaluation are required to assess fracture risk, follow progression of disease and evaluate the effects of treatment. Although there are several risk factors for the prediction of fractures, reduced bone mineral density (BMD) is the strongest predictor. There are several methodologies available for bone mineral measurement. Dual energy x-ray absorptiometry (DXA) has an overall advantage in terms such as precision and radiation exposure. It is a well-established method and has the potential of multi-site measurement. DXA is widely used to assess bone mineral of the spine and hip, however, anomalies of total body DXA have been observed. A Hologic QDR-1OOOW DXA system was used for the investigations. Anomalies in total body bone mineral have been observed during weight change. A total body index (TBI) was therefore developed which related total bone mineral content to stature. In-vitro and in-vivo effects of changes in weight and soft tissue composition and positioning were investigated, using a whole body phantom and a control group. Two groups of anorexic and normal subjects were compared for the TBI, total body BMD and the spine BMD. TBI showed a higher difference between the anorexic (0.7730.11) and normal (1.020.092, p<0.0001) groups than total body BMD and as high as the spine BMD. Precisions of TBI and total body BMD measurements were comparable (1.3%).

THE EFFECTS OF YIGU ZHUANGGU TANG ON BONE MINERAL DENSITY IN PATIENTS WITH GLOMERULAR NEPHRITIS

Beijing Chao Yang Hospital, Affiliate of Capital University of Medical Sciences, Beijing, 100020, P. R. China

OBJECTIVE To study the effects of Yigu Zhuanggu Tang (YZT) on bone mineral density (BMD) and bone mineral content (BMC) in the treatment of glomerular nephritis.

METHODS: BMD and BMC was measured by bone densitometry and serum calcium (Ca) and phosphorus (Pi) was determined before and after treatment. 66 patients with glomerular nephritis (GN) were divided into two groups: (1) Control group: 26 GN patients (9 men, 7 women, average age: 45 years old) were treated with Shenyan Siwei Pian for three months; (2) YZT group: 40 patients (15 men, 25 women, average age: 45.5 years old) were treated with Yigu Zhuanggu Tang for three months.

RESULTS Radius BMD and BMC increased respectively 11.2% and 26.8% in YZT group and increased respectively 0.45% and 4.7% in control group after three month treatment. Radius BMD and BMC in YZT group significantly higher than that in control group (P<0.05). Also the improvement of clinical symptoms was obvious in YZT group (97.2%) compared to control group (42.2%). But there was no significant difference in serum levels of calcium and phosphorus between two groups after treatment.

CONCLUSIONS YZT is an effective Chinese herb for improvement of BMD and BMC in patients with glomerular nephritis.

²Departamento de Enfermería, Escuela Universitaria Ciencias de la Salud, Universidad de Granada

Osteoblasts play an important role during bone tissue development, and are involved in the formation of mineralized bone matrix. Recent reports have suggested that osteoblasts may also exert in bone tissue some activities directly associated with the immune system such as cytokine synthesis and the expression of different molecules involved in antigen presentation and/or T cell activation (HLA-DR, CD44, CD54, CD80, CD86). This suggests that osteoblasts show functional activities similar to those of the phagocytic cells. In this sense, different authors described phagocytic activity of wear debris by osteoblasts, but this activity depends on the chemical composition and size of particles (diameter less than or equal 1 µm).

Different human osteoblasts lines isolated from bone sections obtained during mandibular osteotomy were characterized and cultured. We identified the osteoblasts lines on the basis of their morphological, biochemical and antigenic phenotype. The object of our study was to analyze osteoblasts phagocytic capacity by transmission electronic microscopy. The obtained results (fig. 1) show that osteoblasts can exhibit phagocytic activity to particles of different nature (synthetic polymers and microrganisms) and size (latex beads, Candida albicans and Klebsiella sp). Following exposure to particles the cells incorporated these in the cytosol. No particles were found in the nucleus.

These data can suggest that osteoblasts may have immune function, which may play a role in bone defence against infection.

Fig.1. Photomicrograph of cultured human osteoblasts incubated for 24h with C.albicans.

X±SD	Hip Fracture (42)	Controls (25)	p
Age (years)	79±9	72±8	ns
Weight (Kg)	56±10	63±9	<0.02
Height (m)	1.56±.2	1.53±0.2	ns
Body mass index	23±4	27±3	<0.001
Fat mass (Kg)	20.0±8.4	23.3±7.1	ns
Fat%	34.7±9.1	38.9±7.1	<0.05
Lean Mass (Kg)	33.8±4.3	36.1±4.3	<0.03
BMD (g/cm²)
Total skeleton	0.877±0.1	0.997±0.1	<0.001
Head	1.810±0.2	2.11±0.2	<0.001
Arms	0.670±0.1	0.720±0.1	ns
Legs	0.800±0.1	0.960±0.1	<0.001
Pelvis	0.810±0.1	0.942±0.1	<0.001
Spine	0.924±0.1	1.009±0.1	<0.02

Group	N	Improvement (%) of clinical symptoms and signs
Group	N	ostealgia	reverse body pains	walk pains	lumbal tenderness	lumbal elex-stretch pains
JSZGT	40	65 ± 14	68 ± 15	66 ± 11	62 ± 12	60 ± 11
Caltrate D600	40	38 ± 11	39 ± 11	41 ± 15	36 ± 15	32 ± 15