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Implantation of octacalcium phosphate combined with transforming growth factor-beta1 enhances bone repair as well as resorption of the implant in rat skull defects.

In our previous study, we reported that synthetic octacalcium phosphate (OCP) enhances bone repair if implanted in rat skull defects. We hypothesized that OCP can be used as an effective carrier for transforming growth factor-beta1 (TGF-beta1) to promote bone repair. We designed the present study to investigate histomorphometrically whether combination with recombinant human TGF-beta1 could promote bone repair caused by OCP per se (Control/OCP). A full-thickness standardized trephine defect was made in the rat parietal bone and OCP combined with recombinant human TGF-beta1 (TGF-beta1/OCP) or Control/OCP was implanted into the defect. Four rats from each group were fixed at 2, 4, and 8 weeks after implantation. Histomorphometrical analysis of the percentage of newly formed bone (n-Bone %) and remaining implants (r-Imp %) in the defect was performed. The statistical analysis showed the n-Bone % of TGF-beta1/OCP was significantly higher than that of the Control/OCP in week 4, whereas the r-Imp % of TGF-beta1/OCP was significantly lower than that of the Control/OCP. The present study demonstrated that OCP can be used as an effective carrier for TGF-beta1 and their combination enhances bone repair as well as resorption of the carrier OCP in the early stage of bone formation.

Animals↗

The reactions of articular cartilage to experimental wounding: role of apoptosis.

OBJECTIVE: To determine the cellular and matrix responses to experimental wounding of articular cartilage. METHODS: Immature and mature bovine articular cartilage was used as an in vitro model system to study the cellular responses to cartilage wounding. Explant cultures were wounded centrally with a trephine and maintained for up to 10 days. TUNEL labeling together with ultrastructural analyses were used to assess the nature of the observed cell death. In vitro labeling with 3H-thymidine was used to detect cell proliferation, and 2 antibodies (COL2-3/4M and BC-13) were used to detect changes in matrix turnover. RESULTS: Cell death was observed as a response to wounding and was considered to be a combination of necrosis and apoptosis. In immature tissue, cell death was more pronounced, particularly in the articular surface region. Within the area of cell death, many cells that did not die subsequently underwent proliferation. The collagenous network showed evidence of denaturation in the area of the wound, but "aggrecanase" activity was not detected. CONCLUSION: There are 2 contrasting, but related, responses to cartilage wounding--apoptosis and proliferation. In order to improve cartilage repair, future studies need to elucidate the regulatory mechanisms that determine these responses.

Animals↗

Follow-up examinations including sequential bone marrow biopsies in essential thrombocythemia (ET): a retrospective clinicopathological study of 120 patients.

Diagnosis of essential thrombocythemia (ET) has been usually established by regarding the criteria of the Polycythemia Vera Study Group. Accordingly, a retrospective clinicopathological study was performed on 120 patients with a follow-up ranging between 5 and 13 years and repeated bone marrow trephine examinations. Following the new WHO classification, at presentation patients revealed three distinctive patterns of bone marrow (BM) features: (true) ET in 43 patients, prefibrotic idiopathic myelofibrosis (IMF) in 50 patients, and early IMF in 27 patients. Heterogeneity of morphological features was associated with correspondingly expressed laboratory data. Contrasting initial and early IMF, patients with true ET displayed an about 80% probability to lack splenomegaly, anemia, and increase in the LDH and LAP values and also failed to show any myeloblasts or erythroblasts on the peripheral blood films. Follow-up examinations including sequential BM biopsies (mean interval 39 +/- 31 months) disclosed that of the 43 patients with true ET only one developed an increase in reticulin. On the other hand, 65 of 77 patients with prefibrotic and early IMF evolved into overt myelofibrosis-osteosclerosis. Moreover, survival analysis demonstrated significant differences in our patients. A neglectable proportion of life loss according to a sex- and age-matched general population was found in true ET (less than 11%) opposed to IMF without or mild fibrosis (range 21% to 32%).

Adult↗

Ticlopidine-induced aplastic anemia: two new case reports, review, and meta-analysis of 55 additional cases.

Ticlopidine-induced aplastic anemia (TIAA) is considered very uncommon. We present two new cases, and we review 55 additional cases from the literature. The first case concerns a 70-year-old man who developed severe aplastic anemia 7 weeks after treatment with 500 mg of ticlopidine daily. The patient sustained a severe septic episode, was treated with antibiotics and GM-CSF, and recovered the 14(th) day after ticlopidine withdrawal. The second was an 82-year-old man receiving ticlopidine for 2 years when, during a febrile episode, he was found neutropenic with marrow aplasia. Ticlopidine withdrawal and treatment with antibiotics, transfusions, and G-CSF helped him to recover. When the data of the 57 patients are evaluated, a reversible direct cytotoxic effect of ticlopidine on the pluripotent/bipotent hematopoietic progenitor stem cell is proposed. It is estimated that the real incidence if TIAA is higher, and many cases, initially presented as agranulocytosis +/- thrombocytopenia, might be true aplastic anemias, not proven by marrow aspiration or trephine biopsy. There is no effective monitoring to prevent this side effect. Recombinant growth factors appear not to help in shortening the neutropenic period.

Age Distribution↗

Eosinophilic fibrohistiocytic lesion of bone marrow associated with monoclonal gammopathy and osteolytic lesions.

We describe a 63-year-old male patient with severe osteoporosis, multiple lytic bone lesions, and monoclonal gammopathy (IgG lambda). Whereas the tentative diagnosis in this case was multiple myeloma, bone marrow trephine biopsies of the iliac crest and from an osteolytic lesion of the tibia both showed a peculiar infiltrate consisting of numerous elongated mast cells, eosinophils, and some plasma cells and lymphocytes. The bone marrow lesions fit the diagnosis of eosinophilic fibrohistiocytic lesion of bone marrow (EFHBM). The patient had no abnormality that could be related to a known allergic disease, and no relationship to drug hypersensitivity could be established. The features of the bone marrow infiltrate and its association with monoclonal gammopathy may suggest a linkage between EFHBM and the monoclonal gammopathy.

Bone Marrow Diseases↗

Anaplastic large cell Ki-1 lymphoma involving bone marrow: marrow findings and association with reactive hemophagocytosis.

This report describes the bone marrow findings in four patients whose marrow was involved by anaplastic large-cell Ki-1 lymphoma, an uncommon event in this type of lymphoma. In the marrow aspirate smears, the involvement was subtle, and was in the form of isolated large cells with irregular nuclear configuration, coarse chromatin, prominent nucleoli, and basophilic cytoplasm which might be vacuolated. One case showed paradoxically massive involvement in the trephine biopsy taken from the same site as the marrow aspirate. Reactive histiocytic proliferation with hemophagocytosis was also present. Since marrow aspirate or biopsy may be the first pathologic specimen examined in patients having anaplastic large-cell Ki-1 lymphoma, it is important to be able to recognize the small population of neoplastic cells, which should lead to prompt treatment or further investigations as deemed necessary.

Adolescent↗

Erythropoiesis in primary (idiopathic) osteomyelofibrosis: quantification, PCNA-reactivity, and prognostic impact.

In 64 patients with primary (idiopathic) osteomyelofibrosis (OMF), a morphometric analysis has been performed on bone marrow trephine biopsies following sequential double-immunostaining with monoclonal antibodies against proliferating cell nuclear antigen (PCNA) and erythroid precursor cells (glycophorin C). The purpose of this study was to quantify erythropoiesis and its PCNA-staining capacity and, further, to determine the impact of these parameters for the development of anemia and for prognosis. In comparison with a control group (15 patients), a significant reduction in the number of erythro-normoblasts could be demonstrated, associated with an increase in PCNA-labelling. Moreover, significant correlations between the amount of nucleated erythroid marrow cells and degree of anemia (hemoglobin level, hematocrit, erythrocyte count) and survival could be calculated. Adverse relationships were assessed between number of erythroid cells, thrombocyte count, and spleen size, and also argyrophilic (reticulin/collagen) fiber density. These interactions were thought to reflect the biological behaviour of the disease process, i.e., the progression or extent of myeloid metaplasia. Our findings support ferrokinetic studies suggesting erythroid hypoplasia as one of the major causes of anemia in OMF. The remarkable high PCNA-labelling index of the macrocytic-megaloblastoid appearing erythropoiesis is probably caused by an overexpression of this marker protein. A comparative evaluation of Ki-67 antigen immunostaining in splenic tissue (myeloid metaplasia) and of the PCNA-labelling in pernicious anemia lend support to the assumption of an undue prolongation of the S-phase generated by secondary folate (hematinic) deficiency.

Aged↗

History and development of paleopathology.

In the nineteenth century, accurate descriptive studies, often dealing with pre-Columbian "syphilis," were made by Virchow, J. Jones, and Putnam. Hrdlicka, Moodie, Ruffer, and others in the early twentieth century carried out research on subjects ranging from trephination to schistosomiasis. However, it was not until Hooton, Williams, Stewart, Wells, and others (1930 to 1965) that paleopathology was combined with paleodemography to get a population perspective on health as an equilibrium with disease. Not until recently, with the creative summaries of Jarcho and Brothwell, have we had bone pathologists (Lent Johnson and especially Putschar) and others (Moseley in blood, Cockburn and Allison in epidemiology and mummies) working actively with physical anthropologists and historians. This cooperation has triggered a major revival in the study of bone physiology and responses to disease (Ortner), disease ecology, demography and health, and paleonutrition (Goldstein, Laughlin, Cook and Buikstra, Ubelaker, Angel). New techniques range from histology to immunochemistry and now involve a host of young investigators.

History, 20th Century↗

Surgical treatment of colostomy complications.

One hundred and twenty-three patients (M:F, 0.9:1; mean age 62 years) underwent 156 operations between 1954 and 1984 for correction of late colostomy complications (stenosis 65 patients, prolapse 16 patients, paracolostomy hernia 42 patients). Sixty-three per cent of patients eventually had a good result but in some up to 5 operations were necessary. Local excision of scar tissue at the mucocutaneous junction was associated with a 61 per cent (43/71) success rate for relief of colostomy stenosis. Where local fixation failed to prevent recurrent colostomy prolapse (13/20, 65 per cent of local fixation operations), colectomy and ileostomy was the most effective second procedure (2/3, 67 per cent success rate). Where local repair of a paracolostomy hernia failed (15/32, 47 per cent of local operations), resiting of the stoma to the umbilicus or right side of the abdomen produced better results (3/7, 43 per cent success rate) than resiting to another trephine on the left side of the abdomen (2/14, 14 per cent success rate).

Colostomy↗

Parastomal hernia.

BACKGROUND: Parastomal hernia following formation of an ileostomy or colostomy is common. This article reviews the incidence of hernia, the technical factors related to the construction of the stoma that may influence the incidence, and the success of the different methods of repair. METHODS: A literature search using the Medline database was performed to locate English language articles on parastomal hernia. Further articles were obtained from the references cited in the literature initially reviewed. RESULTS: Parastomal hernia affects 1.8-28.3 per cent of end ileostomies, and 0-6.2 per cent of loop ileostomies. Following colostomy formation, the rates are 4.0-48.1 and 0-30.8 per cent respectively. Site of stoma formation (through or lateral to rectus abdominis), trephine size, fascial fixation and closure of lateral space are not proven to affect the incidence of hernia. The role of extraperitoneal stoma construction is uncertain. Mesh repair gives a lower rate of recurrence (0-33.3 per cent) than direct tissue repair (46-100 per cent) or stoma relocation (0-76.2 per cent). CONCLUSION: The incidence of parastomal hernia is between 0 and 48.1 per cent, depending on the type of stoma and length of follow-up. No technical factors related to the construction of the stoma have been shown to prevent herniation. If repair is required, a prosthetic mesh technique should be considered. Further randomized clinical trials (particularly of extraperitoneal stoma construction) are needed.

Colostomy↗

A case of giant cell myocarditis and malignant thymoma: a postmortem diagnosis by needle biopsy.

This paper reports a case of fulminant giant cell myocarditis arising in association with a malignant thymoma causing death in a 46-year-old woman. Although the diagnosis was suspected in life, postmortem examination was required for confirmation of giant cell myocarditis. Consent was obtained only for percutaneous needle biopsy of the heart. In order to respect the family's wishes and harvest sufficient diagnostic myocardium, a simple needle-based biopsy technique was devised. A bone marrow trephine needle was attached to a 20 ml syringe and, with suction, multiple passes were used to fill 15 tissue cassettes. The cores were placed immediately in formalin and B5 fixatives. High-quality tissue preservation was obtained without crush artefact. Immunohistochemical studies of the biopsy tissue confirmed that the giant cells were of macrophage derivation.

Biopsy, Needle↗

Hodgkin's disease in the bone marrow.

Tumor involvement of the bone marrow in patients with Hodgkin's disease may be suspected in the presence of other manifestations of advanced disease such as fever, lymphopenia, hypoalbuminemia, mixed cellular histologic type, or Stage III or IV disease by other clinical parameters. It occurs more frequently in the older age groups. When anemia, leucopenia, and/or thrombocytopenia are present and are unrelated to recent bone marrow suppressant chemotherapy, marrow involvement is likely to be present. Bone marrow examination, done by multiple trephine biopsies, provides an adequate sampling of tissue and results in a high incidence of detection of involvement by Hodgkin's disease. This manifestation of Hodgkin's disease is associated with a relatively short survival. Aggressive combination chemotherapy is necessary to produce a significant remission.

Adult↗

Migration of A7 immortalized astrocytic cells grafted into the adult rat striatum.

The A7 cell line is an SV40 large T antigen-immortalized astrocyte cell line produced from the neonatal rat optic nerve. Previous studies have demonstrated that A7 cells provide a favorable environment for the survival and growth of cultured neurons and can also stimulate axonal growth after grafting into the rat striatum. The current study was designed to investigate whether A7 cells grafted into adult rat striatum can migrate away from the implantation site. A7 cells were labelled in culture by incorporation of bromodeoxyuridine (BrdU) or by expression of an alkaline phosphatase transgene. The labelled cells were then transplanted into the left striatum of normal adult rats by introducing a blunt-end 22 gauge needle through a trephine hole. The rats were euthanized at periods of up to 30 days after grafting. The A7 cells did not appear to alter the cytoarchitecture of the surrounding brain parenchyma. Labelled A7 cells were observed in both gray and white matter areas, and many were located in areas free of damage due to the implantation procedure. The migration of the BrdU-labelled A7 cells with respect to the implantation needle track was determined on coronal sections. The radial migration distance from the needle tract was similarly determined on horizontal sections. A7 cells migrated progressively longer distances with increasing survival time of the animals: The largest migration distance (1,125 +/- 52 microns) occurred at 30 days after grafting with an estimated migration rate of 31 microns per day. There was no significant directional polarity in the migration of these cells within the striatum. Some of the labelled A7 nuclear profiles were associated with blood vessels, some appeared to be associated with fiber bundles within the striatum, and some were found within the gray matter without apparent association with any anatomical structure. These results demonstrate that A7 immortalized astrocytic cells migrate away from a single implantation site following grafting into the adult rat striatum to populate a large area of the striatum.

Age Factors↗

Validating the boundary element method for forward and inverse EEG computations in the presence of a hole in the skull.

Holes in the skull may have a large influence on the EEG and ERP. Inverse source modeling techniques such as dipole fitting require an accurate volume conductor model. This model should incorporate holes if present, especially when either a neuronal generator or the electrodes are close to the hole, e.g., in case of a trephine hole in the upper part of the skull. The boundary element method (BEM) is at present the preferred method for inverse computations using a realistic head model, because of its efficiency and availability. Using a simulation approach, we have studied the accuracy of the BEM by comparing it to the analytical solution for a volume conductor without a hole, and to the finite difference method (FDM) for one with a hole. Furthermore, we have evaluated the influence of holes on the results of forward and inverse computations using the BEM. Without a hole and compared to the analytical model, a three-sphere BEM model was accurate up to 5-10%, while the corresponding FDM model had an error <0.5%. In the presence of a hole, the difference between the BEM and the FDM was, on average, 4% (1.3-11.4%). The FDM turned out to be very accurate if no hole is present. We believe that the difference between the BEM and the FDM represents the inaccuracy of the BEM. This inaccuracy in the BEM is very small compared to the effect that holes can have on the scalp potential (up to 450%). In regard to the large influence of holes on forward and inverse computations, we conclude that holes in the skull can be treated reliably by means of the BEM and should be incorporated in forward and inverse modeling.

Electroencephalography↗

Frontal bone defect with frontal sinus mucopyocele.

For this patient's treatment, all three consultants advise against the Lynch-type frontoethmoidectomy procedure, with or without mucoperiosteal flap reconstruction of the nasofrontal duct. Treatment plan: Culture and sensitivity of pus; 2-3 weeks of intravenous antibiotics followed by osteoplastic flap fat obliteration of frontal sinus; delayed defect repair with methyl methacrylate (Montgomery). Trephination followed by treatment with local and systemic antibiotics (Donald); removal of infected bone and soft tissue (sinus collapse) and delayed defect repair in 6-12 months (Donald, Calcaterra) with metylmethacrylate (Donald) or in situ cured silicone elastomer (Calcaterra).

Bone Diseases↗

Bone marrow examination in small cell carcinoma of the bronchus: an unnecessary procedure?

One hundred and thirty-seven patients with small cell carcinoma of the bronchus underwent bone marrow trephine and/or aspirate examination as part of their clinical staging. Twenty-four patients (17.5 per cent) were found to have malignant marrow infiltration. In no case was this an isolated finding of metastatic disease, indeed most patients had gross metastatic spread to liver and/or bone. Marrow infiltration has not been shown to be a major prognostic factor in response to chemotherapy or survival in previous studies. We recommend that this staging procedure be discontinued in routine clinical practice, and instead be confined to specific indications in clinical trials.

Bone Marrow Examination↗

Implanted octacalcium phosphate is more resorbable than beta-tricalcium phosphate and hydroxyapatite.

Our previous studies have suggested that synthetic octacalcium phosphate (OCP) could be resorbed and replaced by newly formed bone if implanted in rat skull defects. We hypothesized that the implanted OCP is more resorbable than other commonly used bone graft substitutes of calcium phosphate compounds, such as hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP). To test the hypothesis, the present study was designed to compare histomorphometrically resorption of the implanted OCP, HA, and beta-TCP, which were kept in the experimental cranial defect of rats for a long term. A full thickness of standardized trephine defect was made in the rat parietal bone, and the same volume of granules of OCP, HA, and beta-TCP were implanted into the defect. Five specimens of each group were fixed 6 months after implantation. The percentage of remaining implants (r-Imp%) and newly formed bone (n-Bone%) in the defect was analyzed histomorphometrically. The statistical analysis showed that the r-Imp% of OCP was significantly lower than that of HA and beta-TCP. In contrast, the n-Bone% of OCP was significantly higher than that of HA and beta-TCP. The present study has shown that the implanted OCP in the rat cranial defect is more resorbable than the implanted beta-TCP and HA, whereas the implanted OCP enhances bone formation more than the implanted beta-TCP and HA.

Animals↗

Investigation of a hydroxyapatite and calcium sulfate composite supplemented with an osteoinductive factor.

In the search for a suitable bone graft substitute, a study was conducted using a material that combined a proven osteoconductive composite, hydroxyapatite-calcium sulfate (HA/CS), with an osteoinductive factor, bovine osteogenic factor (OF). The initial study demonstrated the osteoinductive potential of OF in the rabbit muscle model. Once satisfied that the OF was active, it was added to the HA/CS composite and placed in 8-mm trephine defects in the rabbit cranium. This HA/CS/OF was directly compared to HA/CS augmented with a control protein, rabbit serum albumin (RSA). Animals were sacrificed at 4 and 8 weeks. Results demonstrated increased bone formation with the addition of the OF to the composite with bridging of the defects, in most cases, by 4 weeks. No bridging was seen, at this time period, in the other defects left unfilled or filled with HA/CS/RSA. Osteogenic factor, with the appropriate delivery system, can induce bone formation in the rabbit muscle. It may also increase the rate of bone formation at early time periods in a bony defect site when the delivery system is the osteoconductive composite HA/CS.

Animals↗