Burkhard Krempien

Concentration of Transforming Growth Factor Beta in Human Bone Tissue: Relationship to Age, Menopause, Bone Turnover, and Bone Volume

Transforming growth factor β (TGF‐β) is thought to play an important role in bone metabolism, but its relationship to human bone turnover and bone mass has not been examined yet. In this study, we measured the concentration of TGF‐β in 811 samples of male and female bone from four representative sites of the human skeleton and in the supernatants of 72 short‐term human bone marrow cultures from the iliac crest. The concentrations of TGF‐β1 and TGF‐β2 in the bone matrix were positively correlated with histomorphometric indices of bone resorption and bone formation and with serum levels of osteocalcin and bone‐specific alkaline phosphatase. We also observed a positive association between the release of TGF‐β in the bone marrow cultures and serum osteocalcin. Changes in the rate of cancellous or cortical bone remodeling with age or menopause were accompanied by corresponding changes in skeletal TGF‐β. In contrast, there was no significant relationship between the concentration of TGF‐β and bone volume at any skeletal site. In conclusion, our study supports the hypothesis that TGF‐β plays an important role in human bone remodeling, but fails to demonstrate an association between the skeletal concentration of TGF‐β and human bone mass.

Protective effects of a prophylactic treatment with the bisphosphonate 3-amino-1-hydroxypropane-1,1-bisphosphonic acid on the development of tumor osteopathies in the rat: experimental studies with the Walker carcinosarcoma 256.

The present report primarily describes protective effects of a long-term prophylactic treatment with 3-amino-1-hydroxypropane-1,1-bisphosphonic acid (APD) on the development of tumor-induced osteolytic bone destructions. Pretreatment with daily intravenous doses of APD 9.5 mg/kg for 1 week resulted in a significant reduction of Walker carcinosarcoma 256-induced bone destruction, when Walker cells were transplanted intraosseously (2 x 10(6) tumor cells/rat) 7 weeks later. Shorter pretreatment periods (4, 2 or 1 week prior to tumor inocculation) resulted in a nearly total inhibition of bone destruction as well as tumor-induced hypercalcemia. Tumor growth itself was not inhibited by APD pretreatment. Histological and microradiographical findings are reported. Consistent to our experiments and with regard to new immunocytological methods to detect single metastatic tumor cells in the bone marrow, potential risk groups may be defined which may profit from the prophylactic APD treatment to inhibit tumor-induced bone destructions.

Acquired cystic transformation of the kidneys of haemodialysed patients

In the present study, the kidneys of patients who had been on maintenance haemodialysis for variable periods of time were examined at autopsy. In 21 of the 22 patients, multiple pinhead-size to pea-size, nonloculated cysts were observed both in the cortex and the medulla. In some of the cysts (4/20 patients), papillary adenomata were observed which were visible by light microscopy in 3 cases and macroscopically in 1 case. Clinical complications resulting from haemorrhage or neoplastic transformation were not observed in any of the patients of this series. Similar cysts, smaller in size and fewer in number, were also observed in kidneys of uraemic patients who had not been dialysed. Thus, the lesion does not appear to be a specific consequence of maintenance haemodialysis. It appears more likely that extensive cystic transformation of the kidneys of patients in terminal renal failure is made possible by prolonged survival on maintenance haemodialysis. The possibility of malignant transformation of the papillomata cannot be refuted, but epidemiological surveys fail to document more frequent occurrence of renal carcinoma in dialysed patients.

Prophylactic treatment of skeletal metastases, tumor-induced osteolysis, and hypercalcemia in rats with the bisphosphonate Cl2MBP.

Background. The purpose of this study was to investigate the influence of a prophylactic bone protective treatment with the bisphosphonate dichloromethane/diphosphonic acid (Cl2MBP) in an experimental model of osteolysis with intraosseous implantation of the Walker carcinosarcoma 256 B.

Site‐specific human breast cancer (MDA‐MB‐231) metastases in nude rats: Model characterisation and in vivo effects of ibandronate on tumour growth

Animal models are important tools to study the development of bone metastases and to evaluate strategies for their prevention and treatment. We here describe a new model in which tumour inoculation is achieved by injection of cancer cells into the femoral artery. This approach results in the development of multiple osteolytic lesions in the distal femora and proximal tibiae within 18 days after inoculation, with a success rate of 95–100% and no additional comorbidity. In untreated animals, osteolyses expanded continuously at a growth rate of 4.7–8.2 mm2/4 days, causing extensive destruction of resident bone structures by the tumour, significant loss of tibial bone density and a transient rise in urinary bone resorption markers. Continuous daily treatment with ibandronate (10 μg/kg) inhibited further growth of fully established metastases and reduced the mean osteolytic growth rate to 0.03 mm2/4 days. In lesions <6 mm bisphosphonate treatment resulted in a negative growth rate (−0.33 to −0.81 mm2/4 days). When ibandronate was started 3 days prior to tumour cell inoculation, the development of osteolytic lesions was substantially reduced (take rate only 17%) and bone density and structure were mostly preserved. We conclude that the intra‐arterial approach used in this new model of metastatic bone disease results in site‐specific osteolytic lesions with high take rates, steady tumour growth and no additional morbidity. While serial bone marker assessments did not prove useful to monitor osteolytic growth, our studies provide in vivo evidence that ibandronate treatment induces tumour remission by reversal of tumour growth.

Anticancer-agent-linked phosphonates with antiosteolytic and antineoplastic properties: a promising perspective in the treatment of bone-related malignancies?

SummaryBisphosphonates are compounds with a high affinity for bone and other calcified tissues. They inhibit tumor-induced bone destruction and the associated hypercalcemia by hindering the activity of the osteoclasts. Owing to a long biological half-life of bisphosphonates in the bone, a treatment using a prophylactic regimen seems possible. This paper summarizes preclinical studies with the bisphosphonate 3-amino-1-hydroxypropylidene-1, 1-diphosphonic acid and two methyl derivatives; S-N,N-dimethylamino-1-hydroxypropylidene-1,1-diphosphonic acid and 4-N,N-dimetyhlamino-1-hydroxybutylidene-1, 1-diphosphonic acid with respect to their bone-protecting activity in therapy as well as in prophylaxis. To find substances that are useful for the treatment of primary tumor, as well as bone metastasis, we synthesized and testedcis-diammine[nitrilotris(methylphosphonato) (2-)-O1,N1]platinum(II) andcis-diammine{[bis-(phosphonatomethyl)amino]acetato(2-)-O1, N1} platinum(II), which contain both an osteotropic and an antineoplastic moiety. Experiments were carried out: (a) in the intratibial transplanted Walker carcinosarcoma 256B of the rat, which mimics osteolytic bone metastasis, and (b) in the transplantable osteosarcoma of the rat, which shows a histology and metastatic pattern similar to that found in man. These investigations indicate that it is possible to effect adjuvant therapy of bone metastases by combination of two compounds with different properties into one structure without losing the therapeutic characteristics of the parent compounds. They thus provide evidence that it may be possible to design compounds well suited for the therapeutic or prophylactic treatment of bone-related malignancies.

Morphological studies on pathogenesis of epiphyseal slipping in uremic children.

The epiphyseal growth plate of femora (proximal and distal), tibiae, radii and ulnae of seven uremic children were studied to clarify the histopathogenesis of epiphyseolysis. Epiphyseolysis was found to be result of three different processes: (1) growth arrest, (2) excessive erosion of the growth cartilage and of the trabeculae of metaphyseal spongiosa and (3) disturbance of vascularisation of hypertrophic cartilage. By resorptive destruction, secondary hyperparathyroidism causes loss of the chondro-osseous continuity. The ordered trajectoral pattern of the trabeculae in the primary spongiosa is transformed into a dense lace of mechanically inferior trabeculae consisting of woven bone. Impairment of primary mineralization could not be demonstrated. Intensive subperiosteal osteoclastic resorption leads to a reduction of metaphyseal width and to fractures of the unsupported lateral parts of the growth cartilage. There were notable differences between the growth plates in different localisations: in growth plates subjected to axial compression (distal femur, tibia) signs of growth arrest prevailed (reduction of hypertrophic cartilage, occlusion of the growth plate by a transverse plate of bone); in growth plates subjected to shearing forces (upper femur, radius, ulna) epiphyses were seen to slip sideway.

Effects of new bisphosphonic acids on tumor-induced bone destruction in the rat.

SummaryThis report is concerned with therapeutic studies utilizing new bisphosphonic acids on tumor-induced osteolytic metastases. The bone metastases on SD rats were induced by intraarterial and intraosseous transplantation of Walker carcinosarcoma 256B ascites cells. The treatment was carried out using disodium-3-amino-1-hydroxypropylidene-1,1-bisphophonate (APD), diglycidyl-[3-(3,3-bisphosphono-3-hydroxy-propylamino)-2-hydroxypropyl-]urazol-Na2 (DDU) and 1,2,4-triglycidylurazol (TGU). The extent of bone metastases was determined by X-ray on the 5th and 10th days following tumor inoculation, as well as both microradiographically and histologically upon termination of the experiment.High dose DDU produced a clear reduction of the tumor osteolysis, but these positive results were surpassed using APD. The best results were achieved by pretreatment with APD 24 h prior to tumor inoculation.

Skeletal Changes and Growth in Experimental Uremia

Longitudinal growth; bone and growth zone histology; growth cartilage and bone mineralization (tetracycline technique); bone Ca content (neutron activation analysis); bone radiology; serum and urine chemistry; urinary cAMP and serum 25-OH-vitamin D3 were studied in a long-term model of experimental uremia in the rat. Uremia was induced by two-stage subtotal nephrectomy with irradiation of the remaining parenchyma. Ccr in the experimental group was 113 +/- 5.8 micron1/min X 100 g (19.8% of controls) and serum creatinine 1.67 +/- 0.04 mg% (5.1 X control value). Uremic animals were pair-fed with sham-operated controls. In the proximal tibia delayed transformation of cartilage into primary spongiosa with appearance of chondro-osteoid and delayed transformation of primary spongiosa into secondary spongiosa was observed (rickets). Increased amounts of osteoid were present although 25-OH-vitamin D3-levels were high. There were only modest signs of secondary hyperparathyroidism (osteoclast counts; urinary cAMP). In spite of the presence of bone disease, longitudinal growth was not reduced in uremic animals as compared with pair-fed sham-operated animals, but was significantly reduced as compared with ad lib fed control animals. In contrast, weight gain was significantly diminished in uremic animals as compared with pair-fed sham-operated control animals. It is concluded that diminished intake of food is the major determinant of growth retardation in preterminal experimental renal failure.

Pulmonale hyaline Membranen und perinataler Kreislaufschock

1. After death in the perinatal period disseminated intravascular microthrombi can be found extremely often in small vessels of the liver, lungs, adrenals, spleen, and kidneys in the newborn infant (in 82,2% of the examined cases). 2. These microthrombi are considered as the morphological expression of an anoxemic shock in the fetal and neonatal organism, a shock initiated already intra-uterine and extending to the perinatal period. It occurs after various birth complications (abruptio placentae, placenta praevia, abnormal positions of the fetus, prolonged labor, twin pregnancies, diabetes mellitus of the mother and others). Intravascular microthrombi are the pathogenic link between intra-uterine asphyxia, i.e., acidosis and hypoxemia in the small vessels and fetal and neonatal shock. 3. Acidosis and hypoxemia of the fetal and neonatal organism induce aggregation of the thrombocytes and consumption of clotting-factors, hypercoagulability of plasma and disorder of permeability. The formation of pulmonary hyaline membranes in the extravascular space depends on the hypercoagulability of plasma and the disorders of permeability. Pulmonary hyaline membranes must be considered as morphological shock equivalents; they are symptoms of a shock-induced coagulation which provokes the formation of fibrin monomers in the plasma. 4. The deficiency of plasminogen, a characteristic of the immature newborn infant, favours the intravascular polymerization of plasma fibrin monomers to disseminated intravascular microthrombi as well as the extravascular polymerization of fibrin monomers to hyaline membranes of the lungs. 1. Bei der Obduktion perinataler Todesfälle mit pulmonalen hyalinen Membranen lassen sich außerordentlich häufig disseminierte intravasale Gerinnsel in der terminalen Strombahn von Leber, Lunge Nebennieren, Milz und Nieren nachweisen. Im vorliegenden Material fanden sich derartige Gerinnsel in 86,2%. 2. Der Nachweis dieser Mikrothromben wird als morphologischer Ausdruck eines nach verschiedensten Geburtskomplikationen (vorzeitige Lösung der Placenta, Placenta praevia, Lageanomalien, protrahierte Geburten, Zwillingsschwangerschaft, Diabetes mellitus der Mutter u. ä.) auftretenden, bereits intrauterin inszenierten, perinatal fortbestehenden Kreislaufschocks im fetalen und Neugeborenen-Organismus gewertet. Als pathogenetisches Bindeglied zwischen den intravasalen Mikrothromben und dem Kreislaufschock müssen intrauterine Asphyxie, d. h. Acidose und Hypoxämie in der terminalen Strombahn gelten. 3. Acidose und Hypoxämie des fetalen und Neugeborenen-Organismus führen zu Thrombocytenaggregation und Faktorenverbrauch, zu plasmatischer Hypercoagulabilität und Permeabilitätsstörung. Plasmatische Hypercoagulabilität und Permeabilitätsstörungen sind die Voraussetzung für die Ausbildung pulmonaler hyaliner Membranen im Extravasalraum. Pulmonale hyaline Membranen sind in diesem Sinne als morphologische Schockäquivalente, als Ausdruck einer im Schock auftretenden Gerinnungsaktivierung mit Ausbildung plasmatischer Fibrinmonomere zu verstehen. 4. Der für den unreifen Neonatus charakteristische Plasminogen-Mangel begünstigt sowohl die intravasale Polymerisation plasmatischer Fibrinmonomere zu disseminierten intravasalen Gerinnseln, als auch ihre extravasale Polymerisation zu pulmonalen hyalinen Membranen.