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The effects of long-term administration of ethane-1-hydroxy-1, 1-diphosphonate on osteoarthrosis and heterotopic ossification in the mouse knee joint.

Ethane-1-hydroxy-1, 1-diphosphonate (EHDP) was administered weekly for up to 15 months in either a high (50 mg/kg body weight) or low (5 mg/kg body weight) dose to the males of an inbred strain of mouse (SRT/ORT) which suffers from a high incidence of osteoarthrosis of the knee joint accompanied by extensive periarticular soft-tissue metaplasia forming cartilage and bone. The diphosphonate had two effects. In both doses it caused a reduction in the level of bone resorption. In high doses it caused a partial inhibition of mineralization resulting in the deposition of uncalcified osteoid. The diphosphonate did not influence the incidence or severity of the articular degeneration. The development of periarticular sort-tissue metaplasia was not halted by the EHDP although its form was slightly modified in the high dose. In both treated groups the proximal end of the tibia developed into an abnormal club shape as a result of interference with the normal turnover of bony spicules in the metaphyses. High-dose mice given 85Strontium as a tracer for calcium had much lower radioactivity in their bones than did the low dose and control mice.

Aged↗

[Clinical, biological and histological effects of ethane-1-hydroxy 1, 1-diphosphonate (EDHP) in Paget's disease].

The diphosphonates are analogues of the pyrophosphates that are capable of resisting enzymatic destruction and they become fixed in the bone tissue where they have a powerful inhibitory action on osteoclastic resorption and on osteoid mineralization. Ethane-1-hydroxy-1, 1-diphosphonate (EDHP) was administered by mouth at the dose of 20 mg/kg/day over a period of 3 to 6 months to 11 patients with painful Paget's disease. A marked analgesic effect was noted in 7 of the 11 patients. Spectacular metabolic effects were noted with return to normal of the total hydroxyprolinuria/24 h and of the alkaline phosphatases 3 months after treatment in 6 of the 11 treated subjects and a marked improvement in these parameters in 4 other patients. Histologically, there was almost total disappearance of osteoclasia and of osteoblastosis in the affected areas. Clinically, the secondary effects presented in some cases as diarrhoea, and an early and marked rise in phosphoraemia was recorded. Histologically, hyperosteoidosis was seen, equally in the affected and nonaffected zones. This resulted from an almost complete stoppage in the calcification of the osteoid as indicated by the double labelling by tetracycline. The review of the literature shows that considerable therapeutic effects are obtained even with very small doses of EHDP, e.g., 10 mg/kg/day for 3 months, but that these are often accompanied by more marked secondary effects. The diphosphonates appear to be a promising treatment for Paget's disease.

Alkaline Phosphatase↗

[Determination of radiochemical purity of 99mTc(Sn)-methylene diphosphonate using gel chromatography].

The paper presents results of determining radiochemical purity of the osteopathic ligand 99mTc(Sn)-methylene diphosphonate (99mTc(Sn)-MDP). Application possibilities of gel chromatography on Sephadex G 25M were investigated depending on concentrations of reagents and composition of the eluence (Ph = 4-6.5). Concentrations of the ligand were 6.3 x 10(-3) and 6.3 x 10(-1) mol/dm3 Na3 - methylene diphosphonate and of the reductant 4.2 x 10(-1) and 4.2 x 10(-5) mol/dm3 SnCl2. The ratio ligand/reductant remained constant. It was concluded that under the given experimental conditions results of analyses depend solely on the composition of the eluence. The complex partially decomposes if pure saline is used as the eluence. The best results are obtained if the eluence contained both ligand and reductant. According to the obtained results it can be recommended to use eluence with the same concentration of the reagents (excluding technetium), as in the labelled complex. Under this condition Sephadex can be applied in determining radiochemical purity of 99mTc(Sn)-methylene diphosphonate.

Chromatography, Gel↗

Technetium Tc 99m diphosphonate bone scan. False-normal findings in elderly patients with hematogenous vertebral osteomyelitis.

Hematogenous osteomyelitis is usually diagnosed by an abnormal technetium Tc 99m diphosphonate bone scan in symptomatic patients who have positive blood cultures. False-normal 99mTc bone scans have been described recently in neonates with biopsy-proved osteomyelitis. This phenomenon seems to be extremely rare in adults. Two elderly patients with hematogenous vertebral osteomyelitis had normal technetium Tc 99m diphosphonate bone scans when first evaluated. In both cases the bone scans became abnormal four to six weeks after onset of symptoms and two to four weeks after the initial normal results of the study. When suggested by the clinical picture, hematogenous osteomyelitis cannot be ruled out by a normal 99mTc bone scan at any age. Gallium scan, computed tomographic scan, or bone biopsy can be helpful in such cases.

Aged↗

Elimination of phagocytic cells in the spleen after intravenous injection of liposome-encapsulated dichloromethylene diphosphonate. An enzyme-histochemical study.

Dichloromethylene diphosphonate can be used for temporary elimination of macrophages in the spleen when administered after entrapment in liposomes. No comparable effect on the macrophages of the spleen was observed with free dichloromethylene diphosphonate or in the case of empty liposomes. Marginal metallophils on the boundary between white pulp and marginal zone as well as macrophages in the marginal zone and red pulp disappeared from the spleen within one day and remained largely absent for about a week. After this time cells reappeared slowly, and at approximately four weeks after injection their presence in the spleen did not differ from that in control animals. Marginal metallophils and macrophages in the spleen were demonstrated by use of enzyme-histochemical methods and by their capacity to ingest carbon particles.

Acid Phosphatase↗

Technetium 99m diphosphonate uptake and intraosseous hemodynamics during venous congestion in bone. Knee joint tamponade studied in immature dogs.

Venous congestion in bone is a common early feature of inflammatory and degenerative joint diseases. An experimental study was performed of the relationship between the scintigraphic appearance of joints and the intraosseous hemodynamics during venous congestion caused by obstruction of the osseous venous drainage by increased intra-articular pressure. Intra-articular pressure was raised to 75% of mean arterial pressure in one knee each of 8 immature dogs. This caused elevated intraosseous pressure in the distal femoral epiphysis but not in the metaphysis. The elevated intraosseous pressure in the affected femoral epiphysis was associated with decreased technetium 99m diphosphonate uptake and blood flow, unaltered vascular volume and tissue hematocrit, and prolonged transit time of blood in the central cancellous bone. The decreased radionuclide uptake during intraosseous venous congestion thus appeared to reflect a decreased blood supply. However, by multiple regression analysis, the diphosphonate uptake in bone generally varied positively with blood flow and plasma volume and inversely with red cell volume in a nonlinear, multifactorial relationship.

Animals↗

Evaluation of kidney grafts with 99mTc-methylene diphosphonate within 36 hours after transplantation: a marker of ischemic damage.

In order to evaluate renal ischemic damage in transplanted kidneys, renal scintigraphy with 99mTc-methylene diphosphonate was performed in 30 kidney grafts within 36 h after transplantation. The renal uptake was monitored for 100s at 10-min intervals during 90 min following injection. Seventeen patients had an average uptake of radioactivity in the kidney on the 60-, 70-, 80-, and 90-min scintigrams expressed as a percentage of the activity present on the 10-min scintigram of 150% or less (low) and 13 patients had an average uptake of radioactivity of above 150% (high). There was no relationship between the average uptake of radioactivity and warm and cold ischemia times (or the combined value: corrected ischemia), maximal function, reversible rejection, and ureteral obstruction. A significant correlation was found between the average uptake of radioactivity in the kidney and onset of function. Seventy-six per cent of the grafts with a low average uptake of radioactivity had an onset of function on or before day 10 after operation (early) whereas only 15% of the kidneys with a high average uptake of radioactivity had an early onset of function. Twelve per cent of the patients with a low average uptake of radioactivity in the kidneys had their grafts removed for various reasons within 2 months after transplantation, whereas 54% with a high average uptake of radioactivity had their grafts removed. It is concluded that 99mTc-methylene diphosphonate scintigraphy may be useful in the prognostic assessment of graft onset of function and of graft survival.

Adolescent↗

Chemical structure and pharmacokinetics of 99mTc-labelled aminomethane diphosphonic acid derivatives.

99mTc-labelled derivatives of aminomethane diphosphonic acid, general formula: R1R2N-CR3 [PO(OH)2]2, as well as hydroxyethane diphosphonic acid and hydroxypropane monophosphonic acid were investigated with regard to their suitability for skeleton scintigraphy. Bone accumulation was related to chemical structure. It was concluded from the experiments that compounds able to form enols are accumulated to a higher degree and that big substituents influence the affinity to bone negatively by steric hindrance.

Animals↗

Soft-tissue uptake of 99mTc-diphosphonate in systemic AL amyloidosis.

A case of systemic AL (amyloid light-chain-derived) amyloidosis is presented, in which a 99mTc-diphosphonate bone scan demonstrated the intense uptake of tracer in the liver, spleen, heart, thyroid, oropharyngeal region, intestinal tract and uterus. The presence of amyloid deposits in these organs was confirmed at autopsy. A review of the literature is given. In amyloidosis, the binding of radionuclide-labeled calcium-seeking agents, such as diphosphonate and pyrophosphate, may be explained by the high calcium content of amyloid. The presence of calcium in amyloid is due to the presence of the non-fibrillar protein, amyloid P-component (AP). In our patient, the amyloid deposits reacted on immunofluorescence with antibodies directed against AP.

Amyloid↗

Assessment of skeletal uptake of 99mTc-diphosphonate over a five day period.

A new method for the sequential monitoring of skeletal uptake of 99mTc-diphosphonate over a 5-day period, utilising both a gamma camera and whole-body monitor, is presented. It has been found that diphosphonate continues to be removed from bone over this period. Normal subjects appear to lose skeletal activity initially at a more rapid rate than those with altered skeletal metabolism and it is suggested that differences in binding characteristics may exist.

Bone Neoplasms↗

The effects of several diphosphonates on murine bone marrow proliferation.

Six diphosphonates were examined for their ability to alter proliferative responses of mouse bone marrow cells to recombinant human M-CSF and recombinant murine GM-CSF. Risedronate ([2-(3-pyridinyl)-ethylidene] hydroxy bisphosphonic acid) added to in vitro cultures at 10 microM, suppressed the response to M-CSF by 58%, but had no significant effect on GM-CSF-induced proliferation. Ethane-1-hydroxy-1,1-bisphosphonic acid (EHDP), dichloromethylene bisphosphonic acid (Cl2MBP), 3-amino-1-hydroxy-propylidene-1,1-bisphosphonic acid (APD), (4-chlorophenyl)-thiomethylene bisphosphonic acid (tiludronate) and (cycloheptylamino)-methylene bisphosphonic acid (YM-175) had no significant effect. Treatment of mice for 5 or 14 days with 200 mg/kg/day p.o., Cl2MBP or 3 mg/kg/day p.o. of risedronate failed to inhibit M-CSF- or GM-CSF-induced proliferation by recovered bone marrow cells. Addition of Cl2MBP or risedronate in vitro to these cells did not reveal any change in sensitivity to CSFs as a result of exposure to diphosphonate in vivo.

Animals↗

The effect of diphosphonates on periosteal and bone cells in culture.

Calvaria cells were separated into periosteal and bone cells and cultured in the presence of ethane-1-hydroxy-1,1-diphosphonate (EHDP) or dichloromethanediphosphonate (Cl2MDP). Both cell types were affected to the same degree with respect to the effect on cell number and lactate production. The action of the diphosphonates seems therefore not to be specific for one of the cell types.

Animals↗

Comparative study of etidronate and SR 41319, a new diphosphonate, on passive cutaneous anaphylaxis and phospholipase A2 activity.

Diphosphonates are known to affect calcium metabolism. It was therefore felt of interest to investigate the effects of SR 41319, a new Diphosphonate, comparatively to Etidronate disodium (EHDP) on Passive Cutaneous Anaphylaxis (PCA) and on Phospholipase A2 (PLA2) activity which are both mediated by calcium-dependent mechanisms. Results showed that: (1) intravenously, SR 41319 induced a dose-dependent inhibition of PCA more pronounced than the EHDP one; (2) in PLA2 assay, marked inhibition was obtained with SR 41319 whereas EHDP was slightly active. Decrease in Ca entry into mast cells and anti-PLA2 activity of SR 41319 might partly explain the SR 41319 inhibition of PCA.

Animals↗

Splenic uptake of 99mTc-diphosphonate in sickle cell disease associated with increased splenic density on computerized transaxial tomography.

Splenic uptake of 99mTc-diphosphonate was found on the bone scan of a patient with sickle cell disease. No splenic activity was demonstrated on 99mTc-sulfur colloid liver-spleen scans and no evidence of splenic calcification was seen on plain abdominal radiographs. A computerized transaxial tomographic scan, however, did demonstrate a small spleen denser than other abdominal organs. The splenic accumulation of 99mTc-diphosphonate and the increased density on computerized transaxial tomography could be attributed to microscopic splenic calcifications and/or iron deposits in the spleen.

Adolescent↗

Effect of diphosphonates on the synthesis of prostaglandins in cultured calvaria cells.

The calvaria cells cultured for 6 days in the presence of dichloromethanediphosphonate (Cl2MDP) (0.025-250 microM), the synthesis of prostaglandin E2 (PGE2) was inhibited by up to 90%. Inhibition of PGE2 synthesis might be one mechanism whereby this diphosphonate inhibits bone resorption. This effect is not common to all diphosphates since 25 microM 3-amino-1-hydroxypropane-1,1-diphosphonate (AHPDP) stimulated the synthesis of PGE2.

Animals↗

Expanded 99mTc-methylene diphosphonate (MDP) bone scan.

Patients with prostatic cancer frequently require evaluation of bony metastases as well as renal function. 99mTc-methylene diphosphonate, a commonly used bone-imaging agent, is about 60 per cent localized in the bony skeleton and about 40 per cent excreted by the kidneys. Immediate imaging after intravenous injection of the isotope may yield high-quality radionuclide nephrourograms, which provide excellent visual and graphic displays of renal anatomy and excretory function. Our preliminary studies suggest that the immediate 99mTc-methylene diphosphonate scan may usefully expand the value of a routine bone scan to screen for ureteral obstruction. Patients with underlying malignancy who require simultaneous evaluation and follow-up of bony metastases and renal function might be conveniently served by the dual functions of the expanded bone scan to include immediate imaging of the kidneys.

Adenocarcinoma↗

Effects of metallic ions and diphosphonates on inhibition of pericardial bioprosthetic tissue calcification and associated alkaline phosphatase activity.

This study focused on the association of extrinsic alkaline phosphatase (AP) activity with both early and advanced calcification of glutaraldehyde-pretreated bovine pericardial bioprosthetic (GPBP) tissue, and the inhibition of both calcification and AP activity by pre-incubation in diphosphonates (sodium-ethanehydroxydiphosphonate [NaEHDP], aminopropanehydroxydiphosphonate [APD]) and metallic salts (FeCl3, Ga(NO3)3, AICI3). GPBP specimens were implanted subcutaneously in 3 wk old male rats after pre-incubation. Following explantation of the tissue at 72 h and 21 d, calcification was assessed morphologically by light microscopy and chemically by atomic adsorption spectroscopy for calcium content and by molybdate complexation for phosphorus. AP activity was characterized by enzymatic hydrolysis of paranitrophenyl phosphate and by histochemical studies. In both control and pretreated groups, AP levels were greater in 72 h explants than 21 d retrievals, which demonstrated extensive calcification in control explants. All pre-incubations that resulted in inhibition of calcification after 21 d, except for APD, were associated with 72 h AP content which was lower than control specimens. The typical time of initiation of calcification was 72 h, as determined by previous studies with this model system. Covalently bound APD inhibited calcification. Increased AP activity in the APD group may be due to the toxicity of this agent with resultant acute inflammation, or other incompletely understood effects of diphosphonates on calcification and AP. Furthermore, EHDP and Ga3+ incubations were also associated with decreased GPBP AP at 72 h compared to control, but were not effective for inhibiting calcification after 21 d. We concluded that inhibition of peak GPBP AP activity is not necessarily associated with the prevention of GPBP mineralization.

Alkaline Phosphatase↗