Ultrastructural characteristics of bone marrow in patients with hematological disease: a study of 13 cases

There are few transmission electron microscopic studies on bone marrow biopsies of patients with hematological disease owing to the difficulty of overcoming the artifacts of decalcification. Following the fixation of bone marrow biopsies thoroughly before a mild decalcification procedure, ultrastructural studies were performed on 13 patients with varied hematological diseases. Notable features included blood cell disorganization, fibroblast activation, myofibroblast transformation, as well as accumulation of collagen and extracellular amorphous matrix. In addition, excessive blood cell death in leukemia, apoptosis, and macrophage phagocytosis in myelodysplastic syndrome and polycythemia vera, as well as degranulation of eosinophils and megakaryocytes in chronic idiopathic myelofibrosis were predominant, respectively. The observations suggest that polyclonal fibroblast proliferation and extracellular matrix accumulation may result from inflammation resulting from excessive cell death and active material release of blood cells in the bone marrow of patients with hematological disease.     

Ultrastructural Characteristics of Bone Marrow in Patients with Hematological Disease: A Study of 13 Cases

There are few transmission electron microscopic studies on bone marrow biopsies of patients with hematological disease owing to the difficulty of overcoming the artifacts of decalcification. Following the fixation of bone marrow biopsies thoroughly before a mild decalcification procedure, ultrastructural studies were performed on 13 patients with varied hematological diseases. Notable features included blood cell disorganization, fibroblast activation, myofibroblast transformation, as well as accumulation of collagen and extracellular amorphous matrix. In addition, excessive blood cell death in leukemia, apoptosis, and macrophage phagocytosis in myelodysplastic syndrome and polycythemia vera, as well as degranulation of eosinophils and megakaryocytes in chronic idiopathic myelofibrosis were predominant, respectively. The observations suggest that polyclonal fibroblast proliferation and extracellular matrix accumulation may result from inflammation resulting from excessive cell death and active material release of blood cells in the bone marrow of patients with hematological disease.     

Star volume in bone research. A histomorphometric analysis of trabecular bone structure using vertical sections

Conventional bone histomorphometry performed on iliac crest biopsies does not generally provide unbiased stereological estimates of parameters related to bone structure due to the anisotropy of trabecular bone; this, however, can be obtained with vertical sections, which are anisotropic sections, in combination with an anisotropic test system. A practical procedure for obtaining vertical sections from bone is described. The new stereological parameter, the star volume, can provide an unbiased estimation of the absolute mean size of the marrow space and thus give an indirect estimate of the connectivity of trabecular bone structure. The marrow space star volume of vertebral bodies and iliac crest increases with age in both sexes demonstrating that the structural bone changes which occur with age is a topological one with changes in trabecular connectivity. The practical procedure including sampling efficiency for marrow space star volume is described. © 1993 Wiley-Liss, Inc.     

Histopathology of bone marrow reconstitution after allogeneic bone marrow transplantation

In order to study haematopoietic reconstitution in allogeneic bone marrow transplantation we investigated bone marrow histology in 61 biopsies of 37 patients, treated with HLA-compatible bone marrow grafts for leukaemia or severe aplastic anaemia. The biopsies were taken from the day of transplantation until 100 d after transplantation. Stromal changes, in particular oedema, fibrosis and granulomas, were found during the whole period of observation. These changes were more prominent in biopsies from leukaemia patients than from patients with aplastic anaemia. The cellularity in the biopsies increased until 28 d after bone marrow transplantation and was stable thereafter. Initially, only clusters of cells belonging to a single cell lineage were seen, suggesting that the first outgrowth of haematopoietic cells is by proliferation of committed precursor cells. Long-lasting abnormalities in localization of haematopoietic cells in the bone marrow space and of the myeloid: erythroid ratio were seen; dyserythropoiesis was common.     

4248例骨髓活检组织病理的初步分析

对４２４８例骨髓活检组织的病理变化进行分析，结果显示：骨髓活检对许多血液疾病的诊断、预后、分类、分期有一定价值，对早期白血病、慢粒急变，活检可发现先于涂片的变化。分析结果与表明活检对于骨髓纤维化入恶性肿瘤转移等的诊断有较大的作用。并讨论了活检和涂片的优点和不足，指出对有些疾病的诊断，如淋巴瘤、白血病的分类等，两者结合起来分析。     

Polymerase chain reaction in the diagnosis of T-cell lymphoma in paraffin-embedded bone marrow biopsies: a comparative study

AIMS: In routine histological analysis of bone marrow biopsies, the distinction between reactive T-cell infiltrates and T-cell lymphoma can be difficult, even with the use of extensive immunohistochemistry. The aim of this study was to evaluate the diagnostic contribution of TCR-gamma gene rearrangement analysed by PCR. METHODS and RESULTS: The samples studied consisted of 46 paraffin-embedded bone marrow biopsies (diagnosis, staging and follow-up) from 26 patients with T-cell lymphoma. The bone marrow biopsies were categorized into three groups according to the morphological and immunohistochemical results. Group 1, positive for T-cell lymphoma (24 bone marrow biopsies), group 2, suspicion of T-cell lymphoma (15 bone marrow biopsies) and group 3, negative for T-cell lymphoma (seven bone marrow biopsies). DNA could be amplified in 45/46 bone marrow biopsies (98%). Clonal rearrangement was detected in 30/45 bone marrow biopsies tested (67%) including 15/24 bone marrow biopsies (62.5%) of group 1, 11/14 (78.5%) of group 2 and 4/7 (57%) of group 3. In total, PCR analysis supported a diagnosis of T-cell lymphoma in 15/45 bone marrow biopsies (33%), in which histological and/or immunohistochemical examination provided inconclusive evidence of malignancy. CONCLUSIONS: TCR-gamma PCR is a complementary tool for the assessment of T-cell lymphoma in bone marrow biopsies. Optimal evaluation of bone marrow biopsies requires an integrative approach of all available results from morphology, immunohistochemistry, molecular biology and clinical data.     

Bone marrow trephine interpretation: diagnostic utility and potential pitfalls

Good quality paraffin-embedded sections can be obtained from bone marrow trephines, allowing the routine use of these biopsies in diagnostic histopathology. Trephines are superior to aspirates, especially for the assessment of marrow cellularity, the extent and pattern of tumour infiltration and the cell type. Bone marrow biopsies are useful in several areas, e.g. non-haematological disorders, myelodysplasia, the myeloproliferative group, lymphomas and leukaemias, which are briefly discussed below. Further advances in interpretation are likely with more frequent use of bone marrow trephines.     

Role of touch imprint and core biopsy for detection of tumor metastases in bone marrow

A total number of 248 bone marrow trephine biopsies were reviewed and 21 paired biopsies and touch imprints were identified to estimate the role of these two diagnostic methods for detection of tumor metastases from nonhematologic malignancies in the bone marrow. The study period ran between January 1, 1993, and June 1, 1996. Eight histology sections and eight touch imprints were prepared from every case and were reviewed by two pathologists independently. In 20 cases, tumor cells were present without a doubt both on cytologic and histologic preparations. In one touch imprint, single cells were suspicious for malignancy, and the final pathology report was prepared after examination of the histologic sections. There was no positive biopsy in which the imprint was negative for tumor cells, and no positive touch imprint in which the biopsy was negative for such a cells. The results from our study show that every one of these approaches has advantages and disadvantages and that they have a complementary role for identification of solid malignancies in the bone marrow. Diagn. Cytopathol. 1998;18:323–324. © 1998 Wiley-Liss, Inc.     

Value of long-core biopsy in the detection of discrete bone marrow lesions

A series of 256 bone marrow biopsy specimens was obtained at different times from a group of patients with acute myeloid leukaemia, chronic granulocytic leukaemia and acute lymphoblastic leukaemia, and was analysed in parallel with peripheral blood smears and bone marrow aspirate samples. In five of these biopsy specimens the bone marrow lesion was discrete, deeply seated within the marrow cavity and detected only in long-core biopsies. Neither peripheral blood nor bone marrow aspirates obtained at the same time established the correct diagnosis. This suggests that when peripheral blood and bone marrow aspirate samples fail to indicate the diagnosis a long-core biopsy may yield positive results.     

Diagnosis and quantification of bone marrow fibrosis are significantly biased by the pre-staining processing of bone marrow biopsies

AIMS: Marrow fibrosis (MF) is an unfavourable, often lethal complication of haematological neoplasms. Although biopsy technique and staining procedure are standardized, the prestaining processing of bone marrow biopsies (BMBs) varies markedly without any existing data on its significance for the diagnosis of MF. METHODS AND RESULTS: In this study on 712 BMBs from 296 patients with chronic idiopathic myelofibrosis (CIMF), chronic myeloid leukaemia (CML), or healthy bone marrow, MF was a characteristic complication of CML and CIMF. However, diagnosis and quantification of MF and detection of its prognostic significance were significantly biased by fixation, decalcification, embedding, marrow tissue shrinkage during biopsy processing and the thickness of marrow sections (P < 0.000005). The relevance of these influences was explained by their effect on the marrow volume to which the fibre content was related, whereas the stainability of fibres was not affected. Semiquantitative grading of fibrosis and measurements of fibre density could not be adjusted to various methods of processing of bone marrow biopsies (P < 0.003). CONCLUSIONS: Evaluations of MF and its prognostic significance should consider the bias due to the prestaining processing of BMBs and the necessity of an adjustment to the thickness of tissue sections and the degree of marrow tissue shrinkage.     

Infiltrative myelopathy by paracoccidioidomycosis. A review and report of nine cases with emphasis on bone marrow morphology

AIMS: To report nine additional well-defined cases with infiltrative myelopathy by paracoccidioidomycosis (PCM), to describe the specific lesions and infection-related stromal abnormalities, to review the literature on this type of involvement and to introduce a new cause of granulomatous lesions of bone marrow. METHODS AND RESULTS: Different bone marrow specimens were studied (aspirated smears, aspirated clots, biopsy imprints and biopsies) from nine patients with acute or subacute forms of PCM known to have PCM infiltrative myelopathy. CONCLUSIONS: The biopsy specimens were the best for demonstrating bone marrow involvement by PCM. The lesions varied from compact and focal granulomas with few fungal cells to numerous disseminated fungal cells within a loose granulomatous inflammatory reaction, with a continuum between these extremes suggesting a spectrum of immune response to the fungi. Other findings such as bone marrow fibrosis, parenchymal coagulative necrosis and bone necrosis were also observed in the affected areas.     

Fine‐needle aspiration of primary osseous lesions: A cost effectiveness study

Fine‐needle aspiration (FNA) is not widely used in the work‐up of osseous lesions because of concerns regarding its high incidence of nondiagnostic specimens. Although several studies have shown that FNA is less expensive than surgical biopsy, the authors are aware of only one prior study evaluating the cost effectiveness of FNA, which includes the cost of incisional or core needle biopsies necessary to establish a diagnosis when the initial FNA was noncontributory. A computerized search of the pathology records of three medical centers was performed to obtain all FNAs of primary osseous lesions. For each FNA case, all subsequent core needle, incisional or excisional biopsies were recorded as was the result of the definitive operative procedure. The cost of obtaining the definitive diagnosis was calculated for each case including the cost of FNA, imaging guidance utilized, and cost of subsequent surgical biopsy when necessary. The cost of an alternate approach using only surgical biopsy was calculated. The average per patient costs of these two protocols were compared. A total of 165 primary bone tumors underwent FNA. One hundred six of these yielded a definitive cytologic diagnosis. In 59 cases, FNA yielded a result insufficient for definitive therapy necessitating surgical biopsy. FNA investigation of the 165 bone lesions cost 575,932 (average of 3,490 per patient). Surgical biopsy alone would have cost 5,760 per patient. FNA resulted in a cost savings of 2,215 per patient. Diagn. Cytopathol. 2010 © 2009 Wiley‐Liss, Inc.     

Static and dynamic bone changes in hospitalized patients suffering from rickets — a histomorphometric study

Aims : The aim of this study was to assess static and dynamic bone changes in patients suffering from rickets. Methods and results : Transcortical iliac crest biopsies of 15 hospitalized children with rickets were taken after labelling new bone formation with two cycles of tetracycline administration 10 days apart. Undecalcified sections were prepared, appropriately stained and histomorphometric analysis performed. Static and dynamic bone changes were measured including the volume of bone and osteoid, trabecular and cortical bone dimensions and resorptive and mineralization activities. The results were compared with normal values. The nature of the mineralization fronts was noted. Trabecular osteoid volumes of all but one patient was above the normal range of 1.9% (± 0.4%). This patient suffered rickets associated with the Kwashiorkor–Marasmus syndrome. Tetracycline labelling was found to be more sensitive than subjective evaluation of the nature of the mineralization fronts. Despite a balanced hospital diet, a bone formation rate of zero was found in three cases, indicating a need for vitamin D and mineral supplementation. Seven cases had decreased mineralization lag times, indicating response to the balanced diet. Conclusions : This study showed that histometric analysis of labelled bone biopsies is a helpful adjunct to the diagnosis but particularly assessment of response to management of deficiency states in children.     

Diagnostic accuracy and clinical utility of fine-needle aspiration cytology in the diagnosis of clinically primary bone lesions

Optimal treatment of primary bone sarcomas requires minimal disturbance of the tumor prior to preoperative radiation and chemotherapy. Currently, carefully planned incisional or cutting needle biopsies are the favored methods for procurement of specimens. Recently, fine-needle aspiration has gained favor as the initial diagnostic procedure at some centers. We investigated the diagnostic accuracy and the effects of errors in diagnosis and of complications on the patient's course in a series of 101 patients presenting with lesions clinically believed to have arisen in bone. We found that 29% of aspirates were insufficient for the diagnosis; 41% of aspirates yielded a correct diagnosis that had a significant favorable impact on the patient's course, while 20% of aspirates gave a correct diagnosis that did not significantly influence therapy. In 7% of cases, the aspirates were associated with an incorrect diagnosis that negatively influenced therapy, and in an additional 3% of cases an incorrect diagnosis was obtained that had no impact on patient outcome. No complications were encountered in this series of patients.     

Cytology of bone lesions by intraoperative sampling during fracture treatment

Cytology was performed on 314 patients who were treated by surgery for hip joint fracture, to determine and evaluate the role, accuracy, and perspective of intraoperative bone sampling. Specimens were collected from bone lesions during surgery by imprints or driller washing in normal saline. The results were compared with those of subsequent biopsies or clinical follow-up. All 13 neoplastic cases (malignant or benign) were identified by cytology. An accuracy rate of 69.2% was achieved by this method when the type and origin of the neoplasms were to be conclusive. There were no false-positive diagnoses, and all benign conditions showed negative results on cytology (specificity and sensitivity of 100%). Cytology can play a valuable role in the diagnosis of bone lesions. The morphologic diagnostic criteria allow for a high level of diagnostic accuracy of cytologic assessments in most cases of bone lesions, no matter the sampling technique.     

Immunohistology of bone marrow: a modified method of glycol-methacrylate embedding

We describe here a modified glycol-methacrylate embedding technique for bone marrow biopsies which allows high-quality visualization of morphological details and optimal antigen preservation. By the use of this technique it is possible to recognize haemopoietic and stromal cells which are known to play a key role in the regulation of haemopoiesis.     

An appraisal of the value of the bone marrow biopsy in the assessment of proliferative lesions of the bone marrow

The diagnostic value of the bone marrow needle biopsy has proved impressive in a variety of disorders. As a complementary procedure to the aspiration smear it adds an invaluable dimension to the examination of haematopoietic tissue. The procedure is easily learned and safe and should be utilized routinely in haematological practice. The usefulness of the bone marrow biopsy is examined in assessing proliferative lesions of the bone marrow.     

骨髓造血组织切片的形态学观察

采用塑料包理方法，并薄切片，进行ＨＥ、ＨＧＦ、Ｇｏｍｏｒｉ，ＰＡＳ，Ｐｕｓｓｉａｎ Ｂｌｕｅ染色，对１０４例正常人骨髓活检标本进行形态学观测。结果显示了我国正常人骨髓造血组织、脂肪组织、骨小梁、小血管的容量范围及骨髓基质中网硬蛋白，糖元、中性粘多糖、含铁血黄素的分布与含。以期明确我国成人骨髓活检的有关观测标准，为判断骨髓造血组织的病理变化提供依据。     

Histomorphometric assessment of bone turnover in uraemic patients: comparison between activation frequency and bone formation rate

AIMS: The histomorphometric assessment of bone formation rate (BFR/BS) in bone biopsies from uraemic patients is of crucial importance in differentiating low from high turnover types of renal osteodystrophy. However, since BFR/BS relies on osteoblasts, activation frequency (Ac.f), encompassing all remodelling phases, has recently been preferred to BFR/BS. This study was carried out to consider whether estimation of Ac.f is superior, in practical terms, to that of BFR/BS in distinguishing between different rates of bone turnover in uraemic patients. METHODS AND RESULTS: Bone biopsies from 27 patients in predialysis (20 men and seven women; mean age 53 +/- 12 years) and 37 in haemodialysis (22 men and 15 women; mean age 53 +/- 12 years) were examined. The types of renal osteodystrophy were classified on the basis of morphology. Bone formation rate and Ac.f were evaluated according to standardized procedures. The Ac.f was calculated both as a ratio between BFR/BS and wall thickness (W.Th) and as a reciprocal of erosion, formation and quiescent periods (EP, FP and QP). Patients were affected by renal osteodystrophy with predominant hyperparathyroidism (two predialysis and 16 dialysis), predominant osteomalacia (three predialysis and seven dialysis) or that of advanced (nine predialysis and five dialysis) or mild (seven predialysis and four dialysis) mixed type or adynamic type (six predialysis and five dialysis). Activation frequency, which with either formula requires the measurement of W.Th, i.e. the thickness of bone structural units (BSUs), was not calculated in three dialysis patients with severe hyperparathyroidism and in one predialysis and four dialysis patients with severe osteomalacia, because only incomplete BSUs were found. In dialysis, EP was higher in the adynamic than in the other types of osteodystrophy. During both predialysis and dialysis, FP was higher in osteomalacia than in the other forms of osteodystrophy, and in adynamic osteopathy than in hyperparathyroidism or in advanced and mild mixed osteodystrophy. During predialysis and dialysis, QP was higher in the adynamic than in the other forms of osteodystrophy. Correlations were found between BFR/BS and Ac.f, during predialysis (r=0.97) and dialysis (r=0.95). CONCLUSIONS: The superiority of Ac.f in assessing bone turnover, in comparison to BFR/BS, is conceptual rather than practical. The highest values for FP in osteomalacia and for QP in adynamic bone allow a clearer characterization of these low turnover conditions.     

Fine structure of nuclei and cytoplasm of osteoclasts in Paget's disease of bone

To investigate the fine structural features of nuclei and cytoplasm in osteoclasts from patients with Paget's disease of bone, eight bone biopsies were examined. Unusual paracrystalline filamentous formations were found within the nuclei of both actively resorbing and degenerating osteoclasts. These formations were made up of parallel arranged filaments which, in most cases, clustered together regularly. The filaments were slightly argyrophilic and this helped to differentiate them from the surrounding nuclear matrix, even when they were not aggregated in clusters. Argyrophilic filaments were also found loosely aggregated inside the cytoplasm of osteoclasts. In three biopsies two additional and unusual cytoplasmic inclusions were found. The first consisted of irregularly branching, highly argyrophilic membranous strands which were located inside membrane-bound vacuoles. The second consisted of spindle-shaped structures enclosed by two membranes and containing three or more tubules. It is suggested that these results stand as further morphological evidence in favour of the view that a virus, possibly of the paramyxovirus class, has a pathogenetic role in Paget's disease of bone.