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PubMed1994

Fundamentals of bone marrow examination.

Bone marrow evaluation is an important and effective way of diagnosing and evaluating primary hematologic and metastatic neoplasms as well as nonhematologic disorders. Many variations exist for obtaining marrow samples (sites, instruments, techniques); however, the method outlined in this article has proven reliable. Complete evaluation of bone marrow samples should include a brief patient history, pertinent laboratory data, peripheral blood films, bone marrow aspirate smears and sections, and biopsy imprints and sections. Routine examination of the bone marrow as described previously is usually adequate for interpretation. However, application of additional studies using cytochemical, immunocytochemical, immunohistochemical, cytogenetic, and molecular techniques may prove to be of critical importance in the diagnosis of hematologic malignancies.

Biopsy, Needle↗
PubMed1996

Relationship between bone metabolism and plasma cytokine levels in patients at risk of post-transplantation bone disease after bone marrow transplantation.

Bone marrow transplant recipients may carry an increased risk of bone diseases, involving numerous factors that affect bone mineral metabolism. Interleukin-6 is a potent stimulator of bone resorption in vivo. The soluble fraction of interleukin-6 receptor is reported to trigger osteoclast formation by interleukin-6 in vitro. In a cross-sectional study we measured serum bone alkaline phosphatase concentrations and the urinary excretion of pyridinium cross-links in 21 patients after bone marrow transplantation, and investigated the relationship between these values and those for the plasma levels of interleukin-6 and soluble interleukin-6 receptor. Following bone marrow transplantation female--but not male--patients showed higher serum bone alkaline phosphatase values than age- and sex-adjusted controls (p < 0.05). Both female and male patients were characterized by increased urinary excretion values of pyridinium cross-links (p < 0.05). In contrast to a marked increase of interleukin-6 plasma levels (p < 0.001) no significant difference in the soluble interleukin-6 receptor levels was found between patients and apparently healthy persons (p = 0.838). Multiple regression analysis (taking into account different variables of the immunosuppressive regimen applied) revealed the plasma concentration of interleukin-6 as an independent predictor of the urinary excretion of pyridinium cross-links (p < 0.05). In conclusion, in patients following bone marrow transplantation, these findings indicate (a) an increase of bone formation in female--but not in male--patients possibly reflecting primary ovarian failure and (b) an enhancement of bone resorption possibly mediated by circulating interleukin-6.

Adult↗
PubMed2001

High-dose chemotherapy shows a dose-dependent toxicity to bone marrow osteoprogenitors: a mechanism for post-bone marrow transplantation osteopenia.

BACKGROUND: Osteoporosis is a sequela of hemopoietic cell transplantation with a complex multifactorial pathogenesis in which the relative role of chemotherapy and irradiation is not completely understood. Therefore, the authors investigated the toxicity of chemotherapy-only conditioning regimens on bone homeostasis and bone marrow osteoprogenitors, its dose dependency, and the mechanism of chemotherapy-induced osteopenia. METHODS: Fifty-one patients with high-grade non-Hodgkin lymphoma or breast carcinoma who had been treated previously with high-dose + peripheral blood progenitor cell or conventional chemotherapy or who had not received any treatment (prechemotherapy) were enrolled. The authors measured the bone marrow colony-forming unit fibroblast (CFU-f) and long-term culture-initiating cell frequency, forearm bone mineral density, serum osteotropic hormones and metabolic markers of bone formation (plasma osteocalcin), and resorption (urinary collagen I C-crosslinks). RESULTS: Both high-dose chemotherapy regimens caused a 50% reduction in CFU-f frequency, independently of gonadal function status, whereas conventional chemotherapy and prechemotherapy groups were unaffected. Bone mineral density was measured in 26 non-Hodgkin lymphoma patients and again only high-dose chemotherapy caused a 10% loss in cortical bone and 20% in trabecular bone. No endocrine abnormality was found except for the secondary amenorrhea uniformly induced in the high-dose chemotherapy group. In these patients, plasma osteocalcin unexpectedly failed to increase in response to the menopausal increase in bone resorption rate, showing a selective impairment of the osteoblast compartment to cope with increased functional demand. CONCLUSIONS: Chemotherapy without irradiation shows a dose-dependent toxicity to bone marrow stromal osteoprogenitors and can cause osteopenia by direct damage of the osteoblastic compartment, as a mechanism distinct from and summable to hypogonadism.

Adult↗
PubMed2003

Generation of a chimeric mouse reconstituted with green fluorescent protein-positive bone marrow cells: a useful model for studying the behavior of bone marrow cells in regeneration in vivo.

Studies have indicated that bone marrow contains both hematopoietic stem cells and mesenchymal stem cells that can differentiate into a variety of mesenchymal tissues, such as bone, cartilage, muscle, and adipose tissue. Therefore, bone marrow cells are thought to be very useful for cell and gene therapy for various diseases. However, the multipotentiality of these cells remains unclear. To address this issue, we established a chimeric model mouse stably reconstituted with green fluorescent protein (GFP)-marked bone marrow cells. We injected bone marrow cells from GFP-transgenic C57BL/6 mice into the tail veins of recipient wild-type C57BL/6 mice that had been irradiated with a lethal dose of 10 Gy from a cesium source. Microscopic examination and fluorescence-assisted cell sorter (FACS) analysis showed that bone marrow cells, including mesenchymal cells, were almost completely reconstituted with GFP+ cells 5 weeks after transplantation. FACS analysis with lineage-specific antibodies confirmed that the GFP+ cells could differentiate into all types of blood cells. To confirm the usefulness of this mouse model, we studied the role of circulating bone marrow-derived cells in healing of damaged intestine. We performed amputation and anastomosis of the jejunum 10 cm from the pyloric region of the stomach. On the third day after operation, a large number of GFP+ cells were infiltrated in the area of anastomosis, and these cells were positive for CD45 and F4/80 antigens. In 7 days, several cells became negative for CD45 and F4/80 and positive for alpha smooth muscle actin antigen, which is specific for smooth muscle. This finding suggested that bone marrow-derived cells had differentiated into smooth muscle. Because reconstituted bone marrow cells as opposed to injected bone marrow cells, behave naturally, this model is ideal for studying the multipotentiality of bone marrow cells in vivo.

Animals↗
PubMed1994

Detection and significance of bone marrow infiltration at the time of autologous bone marrow transplantation in Hodgkin's disease.

Bilateral iliac crest biopsies were performed at the time of bone marrow (BM) harvest in patients with poor-risk Hodgkin's disease (HD) undergoing autologous bone marrow transplantation (ABMT). 67 consecutive patients with normal trephine biopsies (58 unilateral; nine bilateral) taken 2 weeks prior to harvest were studied. Nine (13%) of patients had BM infiltration with HD on the harvest biopsies. These patients had all previously had only unilateral biopsies. Three patients did not proceed to ABMT because disease progression became clinically apparent, but the remaining six patients were infused with involved marrow. One patient died 6 weeks post ABMT, with no evidence of disease at post-mortem. One patient showed no response to ABMT and four patients either had a partial or complete response to ABMT. Of the responders, three patients are now in complete remission at 53, 39 and 33 months past ABMT.

Adolescent↗
PubMed1986

[Stromal cells of the bone marrow in growing bone].

By means of electron microscopy, cytochemistry and radioautography with 3H-thymidine, the bone marrow stromal cells have been studied in the zones of endochondral osteogenesis in the rabbit and rat femoral bones. In the stromal cells demonstrating a high alkaline phosphatase activity are distinguished: perivascular, reticular fibroblastic, osteogenic cells. Populations of the perivascular phosphatase-positive cells include poorly differentiated DNA-synthesizing forms, as well as cells with signs of differentiation into stromal fibroblasts. Cleft-like spaces in cytoplasm of the fibroblastic reticular cells are, probably, formed as a result of lymphocyte-like mononuclears passing through. Phagocyting stromal elements are presented by macrophages, having perivascular localization and including into composition of erythroblastic islets. Mononuclear macrophages are revealed also on the surface of osseous trabecules, where they participate in destruction of hemopoetic and osteogenic cells.

Alkaline Phosphatase↗
PubMedDate not supplied

[Human bone marrow interferon].

Bone marrow cells from suddenly dying people can produce interferon in response to incoulation of an inducer. The interferon obtained in bone marrow cells did not differ by its properties from the reference preparation. the lots of bone marrow interferon which had been prepared were not inferior in the antiviral activity to leukocyte interferon or were even superior to it. It is suggested that bone marrow cells be used for preparation of human interferon.

Bone Marrow↗
PubMed2001

Successful allogeneic bone marrow transplantation (BMT) by injection of bone marrow cells via portal vein: stromal cells as BMT-facilitating cells.

We examined the importance of the coadministration of bone marrow (BM) stromal cells with BM cells via the portal vein. A significant increase in the number of day-14 colony-forming unit-spleen (CFU-S) was observed in the recipient mice injected with hemopoietic stem cells (HSCs) along with donor BM stromal cells obtained after three to four weeks of culture. Histological examination revealed that hematopoietic colonies composed of both donor hemopoietic cells and stromal cells coexist in the liver of these mice. However, when donor HSCs plus BM stromal cells were administered i.v., neither the stimulatory effects on CFU-S formation nor the hemopoietic colonies in the recipient liver were observed. These findings suggest that the interaction of HSCs with stromal cells in the liver is the first crucial step for successful engraftment of allogeneic HSCs. It is likely that donor stromal cells and HSCs trapped in the liver migrate into the recipient BM and spleen, where they form CFU-BM and CFU-S, respectively.

Animals↗
PubMed2003

Tc-99m methoxyisobutylisonitrile bone marrow imaging for predicting the levels of myeloma cells in bone marrow in multiple myeloma: correlation with CD38/CD138 expressing myeloma cells.

The percentage of myeloma cells in bone marrow is subsequently an important index of disease in patients with multiple myeloma (MM). Bone marrow myeloma cells can be detected by strong CD38/CD138 positivity and light scatter characteristics using flow cytometry. The aim of the study was to evaluate the relationship between the degree of Tc-99m methoxyisobutylisonitrile (MIBI) uptake and the percentage of CD38/CD138 expressing myeloma cells in the bone marrow of patients with MM. A total of 15 patients with MM (mean age: 61.7+/-2.4 years; 7 F and 8 M) were included in the study. Tc-99m MIBI imaging was obtained 20 min after injection of 740 MBq Tc-99m MIBI. Planar spot images of the pelvis and thorax were acquired. The uptake of Tc-99m MIBI in the bone marrow was evaluated using a qualitative and also a semiquantitative scoring system for the bone marrow in areas that included the proximal femurs, anterior iliac crest, and sternum. In all patients, flow cytometry was performed for assessing the percentage of CD38/CD138 expressing myeloma cells in the bone marrow samples. There was a statistically significant positive correlation between the percentage of CD38/CD138 expressing plasma cells in bone marrow and both mean qualitative (r=0.689, p=0.005) and semiquantitative (r=0.669, p=0.006) results of Tc-99m MIBI uptake. In conclusion, our results indicate that increased Tc-99m MIBI uptake of bone marrow is related to the percentage of plasma cell infiltration of bone marrow. Tc-99m MIBI bone marrow imaging may be a useful tool for predicting the levels of myeloma cells in bone marrow of patients with MM.

ADP-ribosyl Cyclase↗
PubMed1993

[Changes of bone marrow MRI pattern in aplastic anemia before and after bone marrow transplantation].

In order to evaluate the usefulness of magnetic resonance imaging (MRI) in estimating bone marrow cellularity, we performed T1-weighted MRI of the lumbar vertebral marrow in two patients with severe aplastic anemia, before and after bone marrow transplantation (BMT). High signal intensity was detected in the bone marrow before BMT, but the bone marrow pattern of MRI became normal after BMT. MRI seems to be effective for evaluating bone marrow cellularity and recovery, and for monitoring therapeutic effects in patients with aplastic anemia before and after treatment with BMT.

Adult↗
PubMed1993

Does in vitro bone marrow purging improve the outcome after autologous bone marrow transplantation?

High-dose therapy with bone marrow rescue is now regarded as an effective treatment for a variety of malignancies. In an attempt to circumvent the scarcity of HLA-matched donors, autologous marrow has been successfully used as the source of cells for hematopoietic reconstitution. There has been considerable controversy as to whether any malignant cells present in the graft could contribute to relapse of disease post-transplant. In this paper we review clinical results obtained by using autologous transplantation and present evidence on how ex vivo tumor purging may contribute to the outcome.

Acute Disease↗
PubMed1993

Gene-marking to trace origin of relapse after autologous bone-marrow transplantation.

Bone marrow harvested for autologous bone-marrow transplantation may contain residual malignant cells even when it is judged to be in remission. Genetic marking and subsequent detection of these cells in recipients would give useful information about the origin of relapse after transplantation. We transferred the neomycin-resistance gene into bone-marrow cells harvested from children with acute myeloid leukaemia in remission. Two patients have relapsed since reinfusion of the marked cells. In both, the resurgent blast cells contained the neomycin-resistance gene marker; thus, remission marrow can contribute to disease recurrence. This method of tracking malignant cells should enable the development of better marrow purging strategies.

Bone Marrow Cells↗
PubMed2000

[Graft of AAA bone combined with autologous red bone marrow for treatment of bone defects].

To evaluate the effect of autolyzed antigen-extracted allogenic bone ("AAA" bone) graft combined with autogenic red bone marrow on bone defects, 58 cases with benign bone tumor(66 long bones) were treated with curettage operation, and the post-curettage bone defects were filled with the complex of "AAA" bone combined with autogenic red bone marrow. Forty two cases(49 bones) were followed up by taking X-ray radiography every 2 months in a period of 1-3 years. The result showed that 49 bones all got bony healing within 9 months(6 months in average). It is suggested that "AAA" bone combined with autogenic red bone marrow may be an ideal material for bone defect repairing.

Adolescent↗
PubMed1989

Haemopoiesis in murine bone marrow and spleen after fractionated irradiation and repeated bone marrow transplantation. I. Erythropoiesis.

Erythropoiesis was studied in mice repeatedly subjected to individual doses of 3 Gy of 60Co gamma-rays at 4-day intervals up to a total dose of 24 Gy on the basis of total bone marrow and spleen cellularity follow-up and analysis of myelograms and splenograms. Half of the mice received 10(6) nuclear cells of syngeneic bone marrow after each fractional radiation dose. It was mainly the spleen which was involved in the adaptation and regeneration of erythropoiesis, its contribution to total erythropoiesis in bone marrow recipients having been as much as 73.9% (day 20 of experiment, total dose 15 Gy). In mice only irradiated, the number of nuclear cells of erythroid lineage decreased to zero values sooner in the spleen (day 16 of experiment, total dose 12 Gy) when compared to the bone marrow (day 24 of experiment, total dose 18 Gy). Analysis of the results of collections made on day 9 after the last irradiation, however, revealed that the haemopoietic microenvironment of the spleen and haemopoietic cells capable of differentiation in the erythroid direction are so resistant to irradiation in only irradiated mice that erythropoiesis in their spleens exhibits signs of regeneration even after the highest total dose of 24 Gy.

Animals↗
PubMed2002

Remyelination of the spinal cord following intravenous delivery of bone marrow cells.

Bone marrow contains a population of pluripotent cells that can differentiate into a variety of cell lineages, including neural cells. When injected directly into the demyelinated spinal cord they can elicit remyelination. Recent work has shown that following systemic delivery of bone marrow cells functional improvement occurs in contusive spinal cord injury and stroke models in rat. We report here that secondary to intravenous introduction of an acutely isolated bone marrow cell fraction (mononuclear fraction) from adult rat femoral bones separated on a density gradient, ultrastructurally defined remyelination occurs throughout a focal demyelinated spinal cord lesion. The anatomical pattern of remyelination was characteristic of both oligodendrocyte and Schwann cell myelination; conduction velocity improved in the remyelinated axons. When the injected bone marrow cells were transfected to express LacZ, beta-galactosidase reaction product was observed in some myelin-forming cells in the spinal cord. Intravenous injection of other myelin-forming cells (Schwann cells and olfactory ensheathing cells) or the residual cell fraction of the gradient did not result in remyelination, suggesting that remyelination was specific to the delivery of the mononuclear fraction. While the precise mechanism of the repair, myelination by the bone marrow cells or facilitation of an endogenous repair process, cannot be fully determined, the results demonstrate an unprecedented level of myelin repair by systemic delivery of the mononuclear cells.

Animals↗
PubMed2005

Defective adhesion and homing of hemopoietic precursors from the bone marrow of patients with myelodysplasia to stromal cells of normal bone marrow cultures.

Hemopoietic precursors from the bone marrow of patients with myelodysplastic syndrome were characterized by lower adhesion to normal stromal sublayer compared to bone marrow precursors from healthy donors, while adhesion to fibroblast monolayer and fibronectin was similar in bone marrow cells from patients and donors. In vitro experiments showed that the percentage of adherent hemopoietic precursors from the bone marrow of patients with myelodysplastic syndrome in normal stromal sublayer and fibroblasts was lower compared to healthy donors. The decrease in adhesive activity of hemopoietic precursors from the bone marrow of patients with myelodysplastic syndrome probably contributes to impairment of cell-cell interactions in the bone marrow of these patients.

Bone Marrow Cells↗