Biomedical subjects

T Nakashima

Publications and source records attributed to T Nakashima.

At least 109 records · Page 6Linked to original sources

PubMed2000

Augmentation of cancellous screw fixation with hydroxyapatite composite resin (CAP) in vivo.

The fixation of fractured bone with screws is important for orthopedic surgery, however, rigid fixation often cannot be attained in elderly patients with osteoporosis. Recently, we developed a new injectable, nonresorbable bone cement (CAP) that possesses mechanical and biological properties superior to those of polymethylmethacrylate (PMMA) cement. CAP can directly bond with bone without intervening fibrous tissue, and the peak curing temperature is 46 degrees C. In this study, we assessed the effects of CAP and PMMA cement on the augmentation of screw fixation in vivo. A cancellous screw was placed in the proximal metaphysis of rabbit tibiae. One side of each tibia was randomly selected to be augmented with CAP or PMMA. The contralateral side received a screw without cement (control). Of the 36 rabbits included in this study, 9 rabbits from each group were sacrificed and the tibial constructs retrieved 1 or 3 months after the initial operation. The screws were then pulled out to failure. The values of the pullout force of the screws augmented with CAP and PMMA were higher than those of the control specimens at both 1 month (319 +/- 58 N for CAP vs. 105 +/- 41 N for control; p < 0.05, 284 +/- 100 N for PMMA vs. 132 +/- 71 N for control; p < 0.05) and 3 months (387 +/- 109 N for CAP vs. 196 +/- 107 N for control; p < 0.05, 372 +/- 145 N for PMMA vs. 242 +/- 100 N for control; p > 0.05) after the operation. However, the average increase in the pullout force between CAP and PMMA augmentation was not statistically significant at either time. The values of energy absorption augmented with CAP and PMMA were also higher than those of the control specimens at both 1 month (129 +/- 54 N*mm for CAP vs. 19 +/- 10 N*mm for control; p < 0.05, 145 +/- 95 N*mm for PMMA vs. 28 +/- 21 N*mm for control; p < 0.05) and 3 months (172 +/- 58 N*mm for CAP vs. 44 +/- 41N*mm for control; p < 0.05, 185 +/- 198 N*mm for PMMA vs. 67 +/- 49N*mm for control; p > 0.05) after the operation. However, there were also no significant differences in energy absorption between the two types of cement augmentation. On the other hand, a significant increase was not observed in stiffness among the CAP, PMMA, and respective control groups at either 1 month (626 +/- 133 N/mm for CAP vs. 441 +/- 180 N/mm for control; p < 0.05, 577 +/- 87 N/mm for PMMA vs. 450 +/- 121 N/mm for control; p > 0.05) or 3 months (622 +/- 144 N/mm for CAP vs. 600 +/- 204 N/mm for control; p > 0.05, 633 +/- 175 N/mm for PMMA vs. 630 +/- 168 N/mm for control; p > 0.05) after the operation, except in the average increase between CAP augmentation and its control 1 month after the operation. These results suggested that a cancellous screw fixation augmented with CAP, as well as PMMA, was effective compared with the unaugmented control in vivo. Because of its biocompatibility and low curing temperature, CAP can be used clinically to augment cancellous screw fixation.

Animals↗

PubMed2000

EGF enhances Ca(2+) mobilization and capacitative Ca(2+) entry in mouse mammary epithelial cells.

Effects of epidermal growth factor (EGF) on the intracellular Ca(2+) ([Ca(2+)](i)) responses to nucleotides, Ca(2+) release from thapsigargin-sensitive stores and capacitative Ca(2+) entry were investigated in cultured mouse mammary epithelial cells. EGF treatment induced proliferation of mammary epithelial cells. We checked for mitotic activity by immunocytochemistry with an anti-PCNA (proliferating cell nuclear antigen) antibody, which stains nuclei of the cells in S-phase of cell cycle. EGF treatment apparently increased the number of PCNA-stained cells compared to those treated with differentiating hormones (insulin, prolactin and cortisol) or without any hormone. Application of EGF did not induce any acute [Ca(2+)](i) response. EGF treatment for 1-2 days in culture, however, enhanced [Ca(2+)](i) responses including [Ca(2+)](i) increase by ATP, UTP and other nucelotides, Ca(2+) release from thapsigargin-sensitive stores, as well as capacitative Ca(2+) entry. Genistein, a tyrosine kinase inhibitor, prevented EGF-induced cell proliferation and the [Ca(2+) ](i) responses in a dose-dependent manner. These results indicate that EGF treatment enhances Ca(2+) mobilization and capacitative Ca(2+) entry, well correlated with cellular proliferation in mammary epithelial cells.

Adenosine Triphosphate↗

PubMed2000

Mechanical evaluation of a bio-active bone cement for total hip arthroplasty.

A new bio-active bone cement, known as CAP, has been developed as an alternative to acrylic bone cement. CAP has improved mechanical properties, with a high modulus that is over five times that of PMMA. The effects of this high modulus are examined by finite element analysis, when the CAP is used in place of PMMA to fix the femoral component in total hip prostheses. The results show a higher tensile stress of 8.76 MPa in the CAP cement, compared with 1.99 MPa in the PMMA cement. However, it is also shown that CAP has a superior fatigue strength of approximately 40 MPa, obtained from a cyclic loading test.

Arthroplasty, Replacement, Hip↗

PubMed2000

Hydrops of placental stem villi complicated with fetal congenital adrenal hyperplasia.

The authors present a case report of hydrops of placental stem villi. Numerous small aechoic spaces were demonstrated by prenatal ultrasonography. The patient spontaneously delivered a female newborn at 26 weeks' gestation. The infant showed hypertrophied clitoris and urogenital sius, and had a normal 46, XX karyotype. Endocrinological examination revealed that 3beta-hydroxysteroid dehydrogenase deficiency caused the anomaly. To our knowledge, this is the first report that congenital genital malformation complicated the placental mesenchymal dysplasia.

3-Hydroxysteroid Dehydrogenases↗

PubMed2000

Development and regression of cochlear blood vessels in fetal and newborn mice.

Various strains of mice have been used for hearing research, but there have been few reports regarding the cochlear vasculature in mice. In this study, the development of the cochlear vasculature was investigated in C57BL/6 mice from day 15 of gestation to day 15 after birth, and mature vessels were also observed in 3-month-old mice. Both India ink injection and the resin casting method were used. On gestational day 17, spiral vessels of the basilar membrane were developing and were elaborating communicating branches that ran toward the external wall and the spiral lamina. On day 18, the spiral vessels showed the largest diameter of all vessels in the cochlea, but these vessels subsequently regressed and finally disappeared by day 14 after birth. The external wall vessels formed a single-layer capillary network at birth and subsequently divided into two layers, which became the vessels of the stria vascularis and the spiral ligament vessels. This process occurred progressively from the basal turn toward the apical turn between days 5 and 8 after birth. A general tendency for the cochlear vasculature to mature from the basal turn towards the apex was observed.

Animals↗

PubMed2000

Methamphetamine-induced striatal dopamine release, behavior changes and neurotoxicity in BALB/c mice.

The behaviors associated with the neurotoxic effects of methamphetamine were evaluated in BALB/c mice. Hyperthermia and behavioral observations were measured 60 min after each subcutaneous injection of methamphetamine (4x4 or 8 mg/kg) or saline, each given 2 h apart. The behavioral observations included stereotyped behaviors, incidence of hemorrhage in breast, salivation and self-injurious behavior (SIB). Repeated administration of methamphetamine produced these behavioral changes and hyperthermia, but resulted in hypothermia by the final injection (8 mg/kg). In addition, the methamphetamine treatment induced a long-lasting dopamine depletion of similar magnitude in the 4 and 8 mg/kg-treated animals. In a time course study striatal monoamine levels were measured 60 min after each injection of these doses. The first and second injections of methamphetamine (8 mg/kg) produced a drastic increase in striatal 3-methoxytyramine; this failed to occur after the third or fourth injection of the same dose. In contrast, 4 mg/kg of methamphetamine also produced an increase in 3-methoxytyramine after the second and third injections of the drug and, in this case, these were maintained for the duration of the treatment. Striatal 3, 4-dihydroxyphenylacetic acid levels also drastically decreased following both doses of methamphetamine, suggesting inhibition of monoamine oxidase in striatum. Moreover, a single injection of methamphetamine increased striatal 2,3-dihydroxybenzoic acid formation. These results suggest that the incidence of hyperthermia, SIB and striatal dopamine neurotoxicity are closely linked to striatal dopamine release and inhibition of monoamine oxidase produced by methamphetamine in BALB/c mice.

3,4-Dihydroxyphenylacetic Acid↗

PubMed2000

A device for removal of femoral distal cement plug during hip revision arthroplasty: a high-powered drill equipped with a centralizer.

A removal procedure of a femoral cement mantle in hip revision arthroplasty has a risk of causing perforation or fracture, especially when removing a well-fixed distal cement plug. A high-powered drill equipped with a centralizer was developed to remove the distal cement plug safely. Using the drill equipped with a centralizer, the cement plug was removed well enough to insert a new component without causing perforation during the operation.

Arthroplasty, Replacement, Hip↗

PubMed2000

Construction and characterization of phage libraries displaying artificial proteins with random sequences.

Three phage libraries, PL1, PL2, and PL3, displaying artificial proteins with random sequences were constructed. The artificial proteins, which are model of ancestral proteins, are derivatives of the 25 kinds of random proteins with about 140 amino acid residues produced via random mutagenesis and combinatorial recombination. The random proteins were displayed on the surface of filamentous bacteriophage as fusion protein with the pIII coat protein at an estimated average number on the phage particles in PL1, PL2, and PL3 of 0.32, 0.32, and 0.08, respectively. Each library was shown to express 10(5) to 10(6) kinds of random proteins. With the phage libraries displaying long random peptides, we now have an effective selection system to observe in vitro evolution of new functional proteins from artificial proteins with random sequences.

Journal Article↗

PubMed2000

Neurological abnormalities in cognitively impaired but not demented elderly.

OBJECTIVE: We determined the prevalence and clinical characteristics of elderly people with cognitively impaired but not demented (CIND) in the community. MATERIAL AND METHODS: We examined 945 elderly residents (aged 65 years or more) in a cross-sectional study of a standardized clinical examination in 27 randomly selected communities in the town of Mifune, a standard farming town in Japan. RESULTS: The prevalence of CIND and dementia were 10.8% and 4.8%, respectively, with both rates increasing with age. The frequency of most neurological signs in CIND elderly was greater than that found in subjects with no cognitive impairment (NCI) and less than that in subjects with dementia. The mean ADL disability score in CIND subjects also lay between the mean values for those with NCI and those with dementia. There were no differences in lifestyles between the CIND group and the other groups. CONCLUSION: The results suggest that a majority of CIND elderly suffer from brain dysfunction due to chronic neurological disorders or aging itself.

Activities of Daily Living↗

PubMed2000

Serum thioredoxin levels as an indicator of oxidative stress in patients with hepatitis C virus infection.

BACKGROUND/AIM: It has recently been suggested that oxidative stress may be associated with hepatitis C virus (HCV) infection. Thioredoxin (TRX) is a stress-inducible thiol-containing protein. The aim of this study was to evaluate the clinical significance of serum TRX levels in patients with HCV-related chronic liver diseases. METHODS: Serum TRX levels were determined with a sandwich enzyme-linked immunosorbent assay kit in 174 serum HCV-RNA positive patients, including 6 asymptomatic carriers, 124 chronic hepatitis, 20 liver cirrhosis, and 24 hepatocellular carcinoma, and in 15 healthy volunteers. RESULTS: The serum TRX levels (medians and [ranges], ng/ml) were significantly elevated in the HCV-infected patients; 30.9 [20.7-37.7] in asymptomatic carriers, 34.5 [8.6-135.6]* in chronic hepatitis, 42.5 [21.4-97.2]* in liver cirrhosis, and 43.9 [11.7-180.3]** in hepatocellular carcinoma (*p<0.05, **p<0.001, vs. 24.9 [1.3-50.7] in healthy controls). Serum TRX levels were significantly correlated with the serum levels of ferritin and fibrogenesis markers, and with the histological stage of hepatic fibrosis. The serum TRX levels before interferon treatment of patients whose serum HCV-RNA was still positive on day 14 following interferon treatment (42.6 [20.1-90.0]) were significantly higher than those of patients whose serum HCV-RNA was negative on day 14 following interferon treatment (25.8 [7.4-59.8], p<0.05). CONCLUSIONS: The serum TRX levels of patients with HCV infection increased with their serum ferritin levels and the progression of liver fibrosis. Patients with higher serum TRX levels exhibited resistance to interferon therapy. Oxidative stress may therefore be responsible for the pathological mechanism of HCV-related liver diseases and be one of the impediments to eradication of HCV during interferon treatment.

Adult↗

PubMed2000

CD4+ T cell-mediated cytotoxicity toward thyrocytes: the importance of Fas/Fas ligand interaction inducing apoptosis of thyrocytes and the inhibitory effect of thyroid-stimulating hormone.

The accumulation of activated CD4+ T cells and antigen (Ag)-dependent cellular interactions between thyrocytes and CD4+ T cells have been determined in thyroid gland from patients with Graves' disease. The Fas/Fas ligand (FasL) interaction between antigen-presenting cells and T cells regulates the apoptosis of the former cells triggered by the latter cells. The inhibition of Fas-mediated apoptosis in thyrocytes could be a underlying mechanism of hyperplasia of thyrocytes in patients with Graves' disease. We investigated the potential role of Fas/FasL interaction between thyrocytes and CD4+ T cells in the induction of Fas-mediated apoptosis of the former cells induced by the latter cells. The presence of only a few specific T cells responsive to a putative autoantigen has hampered the investigation of specific T cell activation toward antigen-presenting cells (APCs). Therefore, we used a superantigen, staphylococcal enterotoxin B (SEB), to examine specific T cell activation toward thyrocytes in vitro since it stimulates a large proportion of T cells with particular Vbeta elements. Spontaneous apoptosis of thyrocytes in culture was not found even in the presence of various kinds of cytokines. In contrast, a clear induction of Fas-mediated apoptosis by anti-Fas IgM was determined in interferon-gamma (IFN-gamma)-stimulated thyrocytes. In addition, a significant cytotoxicity of purified CD4+ T cells toward IFN-gamma-stimulated thyrocytes in the presence of SEB was induced, and the addition of anti-HLA-DR and -DQ monoclonal antibodies (mAbs) or blockade of the Fas/FasL interaction reduced this cytotoxicity. FasL expression of CD4+ T cells cocultured with IFN-gamma-stimulated thyrocytes in the presence of SEB was clearly induced. Furthermore, the addition of mAbs against CD54 and CD58 inhibited both cytotoxicity and FasL expression of CD4+ T cells. The cytotoxicity of CD4+ T cells toward IFN-gamma-stimulated, SEB-pulsed thyrocytes was markedly inhibited when we used thyrocytes cultured with IFN-gamma in the presence of thyroid-stimulating hormone (TSH) as target cells. Our results suggest that 1) CD4+ T cells were activated by thyrocytes expressing MHC class II molecules in an SEB-dependent manner and then expressed FasL. 2) These activated FasL+ CD4+ T cells killed thyrocytes by interacting with Fas on thyrocytes and FasL on activated CD4+ T cells. The presence of costimulating molecules such as CD54 and CD58 on thyrocytes was also necessary to generate activated FasL+ CD4+ T cells. 3) Since the actions of thyroid stimulating antibody (TSAb) toward thyrocytes are similar to those of TSH, one goitrogenic activity of TSAb may, in part, be due to the inhibitory effect on Fas-mediated apoptosis of thyrocytes triggered by activated CD4+ T cells.

Apoptosis↗

PubMed2000

Expression and function of X chromosome-linked inhibitor of apoptosis protein in Sjögren's syndrome.

Apoptotic cell death in acinar and ductal epithelial cells is thought to play an important role in the development of salivary gland dysfunction in patients with Sjogren's syndrome (SS). We examined the expression of anti-apoptotic molecules in salivary glands from patients with SS. The labial salivary glands from six human T-cell leukemia virus (HTLV)-I-seronegative and eleven HTLV-I-seropositive SS patients were analyzed by immunohistochemistry. In vitro experiments were performed with a human salivary gland cell line (HSG cells). Immunohistologic analyses revealed that Bcl-2 and Bcl-x were preferentially expressed in salivary infiltrating mononuclear cells more than acinar and ductal epithelial cells. In contrast, strong X chromosome-linked inhibitor of apoptosis protein (XIAP) expression was evident in both acinar and ductal epithelial cells. The pattern of expression of these anti-apoptotic molecules was similar in both HTLV-I-seropositive and HTLV-I -seronegative SS patients. Western blot analysis confirmed expression of XIAP in cultured HSG cells. The expression of XIAP in HSG cells was increased by IL-1beta, TGF-beta1, or IL-10. However, XIAP expression was down-regulated by TNF-alpha, which induced apoptotic cell death of HSG cells with an increase in caspase-3 activity. These effects of TNF-alpha in HSG cells were antagonized by IL-1beta, TGF-beta1, or IL-10. Our results suggest that XIAP is important in regulating apoptotic cell death of acinar and ductal epithelial cells in patients with SS.

Adult↗

PubMed2000

Properties and pharmacological modification of ATP-sensitive K(+) channels in cat tracheal myocytes.

The effects of levcromakalim and nucleoside diphosphates (NDPs) on both membrane currents and unitary currents in cat trachea myocytes were investigated by use of patch-clamp techniques. In conventional whole-cell configuration, levcromakalim produced a concentration-dependent K(+) current which was suppressed by additional application of 5 microM glibenclamide at -70 mV. When 3 mM ATP was added in the pipette solution, the peak amplitude of the levcromakalim-induced current was much smaller than that in the absence of ATP. When 3 mM uridine 5'-diphosphate (UDP) was included in the pipette solution, much higher concentrations of glibenclamide (>/=50 microM) were required to suppress the 100 microM levcromakalim-induced membrane current in comparison with those in the absence of UDP. In the cell-attached patches, levcromakalim activated a 40 pS K(+) channel which was inhibited by additional application of glibenclamide in symmetrical 140 mM K(+) conditions. UDP (>/=0.1 mM) was capable of reactivating the channel in inside-out patches in which the glibenclamide-sensitive K(+) channel had run down, in the presence of levcromakalim. The K(+) channel reactivated by UDP was suppressed by additional application of either intracellular 3 mM ATP or 100 microM glibenclamide. These results demonstrate that smooth muscle cells in the cat trachea possess ATP-sensitive 40 pS K(+) channels which are blocked by glibenclamide (i.e. K(ATP)) and can be activated by levcromakalim and that intracellular UDP causes a significant shift of the glibenclamide-sensitivity of these channels.

ATP-Binding Cassette Transporters↗

PubMed2000

Nuclear factor-kappaB and caspases co-operatively regulate the activation and apoptosis of human macrophages.

Accumulating evidence suggests that macrophages function as major effector cells in the pathological process of various human diseases. We examined here the role of nuclear factor-kappaB (NF-kappaB) and caspases in the regulation of activation and apoptosis of macrophages. Activation of the human monoblastic leukaemia cell line, U937, by phorbol 12-myristate 13-acetate (PMA) increased the expression of CD14/CD86, and cytokine production. PMA stimulation also increased the expression of both pro-caspase-8 and pro-caspase-3 in U937, but not apoptosis or intracellular caspase-3 activity. PMA also increased the expression of X-chromosome-linked inhibitor of apoptosis protein (XIAP) in U937, suggesting an inhibitory action for XIAP on the caspase cascade in PMA-stimulated U937. Electrophoretic mobility shift assay (EMSA) showed a significant increase of nuclear NF-kappaB activity in PMA-stimulated U937. When a potent NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC), was added to U937 cell culture in the presence of PMA, apoptosis was triggered by activation of caspase-3, which was induced by caspase-8 activation. XIAP expression was markedly suppressed in PMA-treated U937 in the presence of PDTC. The inhibitors of caspase-8 and caspase-3 mostly inhibited apoptosis of U937 treated with PMA in the presence of PDTC. Furthermore, a phenotype of U937 treated with PMA and PDTC in the presence of caspase inhibitor was almost identical to that of unstimulated U937. Our results suggest that the signalling pathways involved in the activation and apoptosis of human macrophages could be co-operatively regulated by the use of NF-kappaB and caspase inhibitors, thus enabling the control of macrophage function and number.

Apoptosis↗

PubMed2000

Serum total bile acid level as a sensitive indicator of hepatic histological improvement in chronic hepatitis C patients responding to interferon treatment.

BACKGROUND AND METHODS: Serum total bile acid (TBA) levels are used clinically as a sensitive and reliable index of hepatobiliary diseases. In the present study, to assess the clinical usefulness of determining TBA in interferon (IFN)-treated patients, changes in liver function test values, including TBA and liver histology, were examined in 36 chronic hepatitis C patients for 3 years after a sustained response to IFN treatment. RESULTS: Alanine aminotransferase and gamma-glutamyl transpeptidase values significantly decreased during the period of IFN treatment compared to the same measures before IFN treatment. Albumin, cholinesterase, total cholesterol and platelet count values temporarily decreased during IFN treatment, then increased significantly and reached a plateau 6-12 months after the end of IFN treatment. The zinc sulphate turbidity test and TBA values began to decrease during IFN treatment and continued to decrease during the 3-year follow-up period. The histological activity index of the liver (Knodell's score) significantly decreased, whereas the staging score was unchanged 1 year after the end of IFN treatment. In patients who had a TBA value > 10 micromol/L before IFN treatment, a significant correlation was observed between the decrease of TBA and liver histology grading score. CONCLUSIONS: A decrease in serum TBA level reflected histological improvement in the liver more precisely than did changes in the other liver function test values following IFN therapy.

Adult↗

PubMed2000

Clinicoepidemiologic features of sudden deafness diagnosed and treated at university hospitals in Japan.

Nationwide epidemiologic surveys were done 3 times by the Research Committee of the Ministry of Health and Welfare in Japan to investigate the number of patients and clinical manifestations of idiopathic sudden sensorineural hearing loss (sudden deafness). The first, second, and third surveys were carried out during the 1970s, 1980s, and 1990s, respectively. For each patient, we have compiled data regarding age, sex, onset day, presence or absence of vertigo, and hearing ability, including initial and final audiograms, in a computer in the Nagoya University Computer Center. Regarding patients who visited university hospitals, complete data were collected for 421 subjects (age: 40.2+/-15.4 years, female: 203, male: 218) between July 1973 and June 1974, for 813 subjects (age: 45.1+/-15.4 years, female: 407, male: 406) in 1987, and for 1112 subjects (age: 49.1+/-16.0 years, female: 591, male: 521) in 1993. The number of patients with sudden deafness treated in university hospitals in Japan has increased, especially in the elderly population. This increase is associated with population increase and age-adjusted incidence rates per 100,000 in the elderly population. The initial and final hearing levels in the first survey were worse than those in the second and third surveys; there was no significant difference in hearing between the second and third surveys. The hearing levels were worse in children and elderly patients than in the other age groups.

Adolescent↗

PubMed2000

Effect of vitamin K2 on osteoblast apoptosis: vitamin K2 inhibits apoptotic cell death of human osteoblasts induced by Fas, proteasome inhibitor, etoposide, and staurosporine.

Vitamin K2 is used for the treatment of osteoporosis, but the precise mode of action is still not clear. We investigated the effects of vitamin K2 on apoptosis of human osteoblasts. Human osteoblastic cell line MG63 cells and human primary osteoblast-like cells obtained from bone fragments in corrective surgery were used as human osteoblasts. Cells were cultured with or without various concentrations of vitamin K2 and tumor necrosis factor-alpha (TNF-alpha). We then determined the proliferative response, expression of Fas and Bcl-2-related proteins, and Fas-mediated apoptosis of these cells induced by anti-Fas immunoglobulin M (IgM). In addition, the effect of vitamin K2 in osteoblast apoptosis induced by Z-Leu-Leu-Leu-aldehyde (LLL-CHO), etoposide, or staurosporine was also examined. Human osteoblasts did not show spontaneous apoptosis in culture, even in the presence of vitamin K2 or TNF-alpha. Furthermore, proliferation of the cells was not influenced by vitamin K2 or TNF-alpha. Fas was functionally expressed on human osteoblasts, and the treatment with TNF-alpha significantly enhanced both Fas expression and Fas-mediated apoptosis of osteoblasts. The addition of vitamin K2 to the culture resulted in a dose-dependent inhibition of functional Fas expression on osteoblasts, in the presence or absence of TNF-alpha. Treatment of human osteoblasts with vitamin K2 clearly suppressed Bax expression of the cells, although the expression of Bcl-2 was not influenced by vitamin K2. Fas ligand (FasL) cDNA transformants were cytotoxic against osteoblasts, and the cytotoxicity was increased when osteoblasts were treated with TNF-alpha. The addition of vitamin K2 to osteoblasts significantly decreased the cytotoxic effects of FasL cDNA transformants. Furthermore, apoptosis of human osteoblasts induced by LLL-CHO, etoposide, or staurosporine was also clearly suppressed in vitamin K2-treated osteoblasts. Our results suggest that vitamin K2 inhibits apoptotic cell death of osteoblasts and maintains the number of osteoblasts. These actions may explain the therapeutic efficacy of vitamin K2 in osteoporosis.

Apoptosis↗

PubMed2000

Etidronate inhibits human osteoblast apoptosis by inhibition of pro-apoptotic factor(s) produced by activated T cells.

Humoral factors produced by activated T cells are thought to be important in the development of bone loss in patients with rheumatoid arthritis (RA). We investigated the inhibitory effect of etidronate disodium (EHDP) on apoptosis of human osteoblasts induced by supernatants from in vitro activated T cell cultures. Human osteoblastic cell line MG63 cells and human primary osteoblast-like cells were used in the present study as human osteoblasts. T cells were incubated with interleukin-2 and further activated with 1 2-o-tetradecanoyl-phorbol 13-acetate and ionomycin, either in the presence or absence of EHDP. After we carried out the cultivation, we examined the cytotoxicity of cultured T cell supernatants toward MG63 cells and human primary osteoblast-like cells. Supernatants from activated but not resting T cell cultures efficiently induced apoptosis of MG63 cells and primary osteoblast-like cells. Supernatants from activated T cell cultures, incubated with EHDP, exhibited significantly less cytotoxicity than did supernatants incubated in the absence of EHDP. In contrast, the cytotoxicity of activated T cell culture supernatants was not affected by direct treatment of human osteoblasts with EHDP. The concentration of soluble Fas ligand in activated T cell culture supernatants was actually increased by EHDP. However, EHDP did not influence soluble Fas and tumor necrosis factor-alpha concentrations in the supernatant. Furthermore, treatment of human osteoblasts with EHDP did not alter their expression of Bcl-2/Bcl-xL or their sensitivity to anti-Fas immunoglobulin M-induced apoptosis. Our results suggest that EHDP inhibits the production of soluble factor that induces apoptosis of human osteoblasts and thus exhibits a protective action toward human osteoblast apoptosis induced by activated T cell culture supernatants. Although the exact EHDP-regulated molecule that induces apoptosis of human osteoblasts is unknown at present, our study may explain part of the therapeutic action of bisphosphonates in RA complicated by bone loss.

Apoptosis↗