Effects of air tourniquet on the antibiotics concentration,in bone marrow,injected just before the start of operation

Infection is one of the most serious complications after artificial arthroplasty. In order to establish the effective prevention for after operative infection, we measured the serum and bone marrow blood cefmetazole (CMZ) concentration time dependently (1 g CMZ, one shot). Furthermore, we studied the effect of air tourniquet on CMZ transmit into bone marrow blood. Thirteen knees with total knee arthroplasty (TKA) were included in the study. As a control group, 11 hips with total hip arthroplasty (THA) were also included. In TKA, air tourniquet was used during operation in all cases. Just before the start of the operation, 1 g CMZ was injected intravenously (one shot). Subsequently we sampled peripheral blood and bone marrow blood time dependently. Cefmetazole concentration was measured with HPLC. In the THA group, serum and bone marrow blood CMZ concentration showed almost the same time-dependent change. On the other hand, in the TKA group we could not detect CMZ in bone marrow blood in cases where CMZ was injected within 8 min before starting use of an air tourniquet. If CMZ was injected more than 10 min before starting use of the air tourniquet, CMZ concentration in bone marrow blood was much lower than minimum inhibitory concentration (MIC) for Staphylococcus aureus; but after releasing the air tourniquet, CMZ concentration in bone marrow blood was higher than MIC for S. aureus. These data suggested that our injection method is effective for prevention of infection both during and just after operation in the THA but in the TKA, CMZ should be injected more than 10 min before starting to use the air tourniquet.     

The bone marrow aspirate of healthy subjects

Bone marrow aspirates were obtained from the right or left posterior superior iliac spine of 50 healthy volunteers, 30 men and 20 women. Reference ranges were derived for each cell type and for the myeloid : erythroid (M : E) ratio. The M : E ratio and the percentage of neutrophils were significantly higher in the women and the erythroid component significantly lower. All 28 evaluable men and 11/17 evaluable women had storage iron present in more than trace amounts. The percentage of erythroblasts with detectable iron granules varied very widely, from 3% to 69% in those with more than a trace of storage iron. Minor dyserythropoietic features were present in a high percentage of subjects and 19/50 subjects had one or two dysplastic megakaryocytes. Granulocytic dysplasia was not detected.     

Y-body study in bone marrow precursors, peripheral blood cells and alveolar macrophages for demonstration of haemopoietic engraftment in allogeneic bone marrow transplantation

The value of Y-body study for assessment of haemopoietic engraftment was analyzed in 50 consecutive patients submitted to allogeneic bone marrow transplantation (BMT) (sex-matched in 28 cases, sex-mismatched in 22). The study was performed weekly on bone marrow and peripheral blood smears in all cases, and alveolar macrophages were also studied in 15 patients in whom bronchoalveolar lavage was carried out because of concurrent respiratory disturbances. The analysis was performed blindly by 2 independent observers. In both sex-matched and sex-mismatched cases there was an absolute concordance between recipient and donor Y-body results, as well as with the simultaneous cytogenetic study. The engraftment of erythroid and granulopoietic lines was documented at day +14 in all cases of sex-mismatched BMT, whereas megakaryocyte and lymphocyte take was demonstrated at d +21. On the other hand, the results from alveolar macrophages were in accordance with those obtained in the simultaneous study of bone marrow precursors after BMT. The above results indicate that Y-body analysis is a simple and useful tool for the demonstration of bone marrow take in sex-mismatched BMT.     

The effect of granulocyte colony-stimulating factor administration in healthy donors before bone marrow harvesting

To investigate whether granulocyte colony-stimulating factor (G-CSF) administration to donors before harvest may lighten the burden imposed on them and accelerate the bone marrow (BM) recovery, we administered 2 microgram/kg/d of G-CSF for five consecutive days before the marrow harvest. All of the donors tolerated the G-CSF administration well without severe adverse events. After 5 d of G-CSF treatment, CD34+ cells and granulocyte-macrophage colony-forming units (GM-CFU) in the donors' BM exceeded baseline values by 4.2-fold (range 0.71-316) and 1.6-fold (0.28-118) respectively. The concentration of total nucleated cells (x 107/ml) in the graft increased from 1.61 (0.95-3.23) to 2.44 (1.27-4.01). Although we collected 1020 ml of BM and obtained 1.50 x 1010 nucleated cells from unprimed donors, 940 ml of BM were sufficient to obtain 2.14 x 1010 nucleated cells from primed donors. However, G-CSF-primed BM did not shorten the time to tri-lineage engraftment and the duration of hospitalization compared with unprimed BM, although primed BM contained more CD34+ cells than baseline values. We consider that the advantages of BM priming are not the acceleration of BM recovery but rather the reduction of blood loss during BM harvesting.     

Effects of a preformed extracellular matrix on long-term serum-free bone marrow culture

Summary The extracellular matrix (ECM) produced by the stromal layer plays a key role in the regulation of commitment and differentiation of hematopoietic cells. Long-term bone marrow culture (LTBMC) allows analysis of the stromal microenvironment. Recently, serum-free LTBMC has been described, but the formation of a classical adherent layer was never observed under these conditions. We have evaluated the effect(s) of a chemically well defined ECM on serum-free and serum-dependent LTBMC. In serum-dependent cultures ECM did not induce a significant increase of hematopoiesis. In serum-free conditions, a marked improvement of hematopoiesis was observed, both in terms of CFU-GM and BFU-E yield and in duration of cultures. A confluent stromal layer was observed only in the presence of ECM. The present results indicate that the addition of ECM to serum-free cultures provides a standardized culture condition, while improving progenitor cell recovery and allowing formation of a confluent stromal layer. Moreover, ECM+ LTBMC may provide a model to study the effect(s) of adhesive proteins and hematopoietic growth factors normally present in serum.This work was supported by an AIRC grant and the Program Terapia dei Tumori,Istituto Superiore di Sanità, Rome     

The dynamics of adult haematopoiesis in the bone and bone marrow environment

This review explores the dynamic relationship between bone and bone marrow in the genesis and regulation of adult haematopoiesis and will provide an overview of the haematopoietic hierarchical system. This will include the haematopoietic stem cell (HSC) and its niches, as well as discuss emerging evidence of the reciprocal interplay between bone and bone marrow, and support of the pleiotropic role played by bone cells in the regulation of HSC proliferation, differentiation and function. In addition, this review will present demineralized bone matrix as a unique acellular matrix platform that permits the generation of ectopic de novo bone and bone marrow and provides a means of investigating the temporal sequence of bone and bone marrow regeneration. It is anticipated that the utilization of this matrix‐based approach will help researchers in gaining deeper insights into the major events leading to adult haematopoiesis in the bone marrow. Furthermore, this model may potentially offer new avenues to manipulate the HSC niche and hence influence the functional output of the haematopoietic system.     

Effects of obstructive jaundice on neutrophil production and acquisition of chemotactic activity in the bone marrow

Background and Aims:  Increased numbers and enhanced functions of peripheral neutrophils have been observed in obstructive jaundice. However, the effects of obstructive jaundice on the bone marrow, that is neutrophil production and acquisition of neutrophil chemotactic activity, have been poorly understood. In the present study, differentials of bone marrow cells and chemotactic activity of bone marrow neutrophils were evaluated in bile duct-obstructed rats.
Methods:  Male Wistar rats underwent either bile duct obstruction for 10 days or bile duct obstruction for 4 days followed by 6 days' internal biliary drainage. Differentials of peripheral blood and bone marrow cells were sequentially determined. Chemotactic activity of peripheral and bone marrow neutrophils was evaluated with a modified Boyden method using interleukin-8 (recombinant rat Gro-β) as a chemoattractant.
Results:  Numbers of peripheral neutrophils significantly increased after bile duct obstruction. Significant increases in the myeloid/erythroid (M/E) ratio of bone marrow cells were observed after bile duct obstruction. The neutrophil proliferative pool (promyelocytes and myelocytes) increased initially, followed by an increased neutrophil storage pool (metamyelocytes, bands, and segmented neutrophils). The M/E ratio as well as the neutrophil proliferative and storage pools normalized after internal biliary drainage. Chemotactic activity was enhanced in both peripheral and bone marrow neutrophils after bile duct obstruction, and enhanced chemotaxis was alleviated with internal biliary drainage.
Conclusion:  The present results strongly suggest the principal role of the bone marrow in increasing the number of neutrophils and their chemotactic activity during obstructive jaundice.     

Effects of obstructive jaundice on neutrophil production and acquisition of chemotactic activity in the bone marrow

BACKGROUND AND AIMS: Increased numbers and enhanced functions of peripheral neutrophils have been observed in obstructive jaundice. However, the effects of obstructive jaundice on the bone marrow, that is neutrophil production and acquisition of neutrophil chemotactic activity, have been poorly understood. In the present study, differentials of bone marrow cells and chemotactic activity of bone marrow neutrophils were evaluated in bile duct-obstructed rats. METHODS: Male Wistar rats underwent either bile duct obstruction for 10 days or bile duct obstruction for 4 days followed by 6 days' internal biliary drainage. Differentials of peripheral blood and bone marrow cells were sequentially determined. Chemotactic activity of peripheral and bone marrow neutrophils was evaluated with a modified Boyden method using interleukin-8 (recombinant rat Gro-beta) as a chemoattractant. RESULTS: Numbers of peripheral neutrophils significantly increased after bile duct obstruction. Significant increases in the myeloid/erythroid (M/E) ratio of bone marrow cells were observed after bile duct obstruction. The neutrophil proliferative pool (promyelocytes and myelocytes) increased initially, followed by an increased neutrophil storage pool (metamyelocytes, bands, and segmented neutrophils). The M/E ratio as well as the neutrophil proliferative and storage pools normalized after internal biliary drainage. Chemotactic activity was enhanced in both peripheral and bone marrow neutrophils after bile duct obstruction, and enhanced chemotaxis was alleviated with internal biliary drainage. CONCLUSION: The present results strongly suggest the principal role of the bone marrow in increasing the number of neutrophils and their chemotactic activity during obstructive jaundice.     

Association of bone marrow lesions with knee structures and risk factors for bone marrow lesions in the knees of clinically healthy, community-based adults

OBJECTIVES: Subchondral bone marrow lesions (BML) are involved in pain and progression of knee osteoarthritis (OA). Little is known about their role in the knee in those without clinical OA. Our aim was to examine the prevalence and risk factors for BML, and their relationship with other knee structures in community-based adults without clinical OA. METHODS: Two hundred ninety-seven healthy subjects without knee pain or injury were recruited from an existing community-based cohort recruited at baseline in 1990-1994. Subjects with a single magnetic resonance imaging (MRI) of their dominant knee at follow-up were studied in 2003-2004. BML, cartilage defects, cartilage volume, and bone area of the knee were assessed using MRI. RESULTS: Thirty-nine subjects (13%) had evidence of BML. BML were associated with the presence of cartilage defects in the medial (odds ratio (OR) 1.80, P = 0.004) and lateral (OR 1.45, P = 0.04) tibiofemoral compartments, but not cartilage volume. BML were positively associated with total tibial bone area (OR 1.22, P = 0.02). Increasing age (OR 1.10, P < 0.001), male gender (OR 3.86, P = 0.01), and increasing body height (OR 1.07, P = 0.03) were independently associated with BML in the total tibiofemoral compartment. CONCLUSIONS: BML are present in the knees of community-based adults without clinical OA and are strongly associated with tibiofemoral cartilage defects. Risk factors for BML were age, male gender, and body height. Longitudinal studies will be needed to clarify the role of BML in structural change of the knee and how this relates to the pathogenesis of symptomatic knee OA.     

骨髓活检与涂片同步观察对诊断慢性淋巴细胞白血病的探讨

目的 :探讨骨髓活检和骨髓涂片同步观察在慢性淋巴细胞白血病 (CLL)诊断方面的重要性。方法 :针对 2 0例CLL患者 ,采用骨髓抽吸 活检双标本一步法取材 ,同时观察其骨髓涂片和活检塑料包埋切片。结果 :①骨髓活检切片增生度检出率为 10 0 % ,骨髓涂片检出率为 70 % ,两者差异有显著性意义 (P <0 .0 5 ) ;②切片中原始和幼稚淋巴细胞百分率略高于涂片 ;③ 85 %CLL患者伴有网状纤维增高 ,5 %CLL患者有胶原纤维增高。结论 :同步观察比常规涂片形态学观察的检出率、准确率均明显增高。     

Comparative evaluation of bone marrow aspirate particle smears, biopsy imprints, and biopsy sections

Bone marrow aspirate particle smears, biopsy imprints, and biopsy sections were compared to determine the accuracy of the three samples in assessing for overall cellularity, differential cell count, megakaryocyte density, iron stores, and tumor infiltration. Aspirate particle smears and biopsy imprints were stained by Wright-Giemsa method. Aspirate particle smears were also stained with Prussian-blue. Biopsy sections were 1 1/2-2 micron thick and were prepared from non-decalcified plastic embedded samples and stained with combined Prussian-blue-hematoxylin-eosin, and Giemsa. One hundred-eight sets of specimens from 99 patients were examined. In 20 cases, chi-square analysis showed a comparable degree of cellularity (p less than 0.001) and megakaryocyte density (p less than 0.001) among the three preparations. Differential count comparison by regression analysis indicated that mean percentages of neutrophilic cells in the proliferation compartment were comparable in the three groups (p less than 0.01). A better correlation was obtained among the three groups in the percent neutrophilic cells in the maturation-storage compartment, normoblasts, eosinophils, and plasma cells (p less than 0.001). Lymphocytes in the aspirate smears correlated with the biopsy imprints (p less than 0.01) but not with the biopsy sections (p greater than 0.05). Monocytes did not correlate in any of the groups (p greater than 0.05). In 47 cases, chi-square analysis of iron stores in the aspirate particle smears correlated well with those in the biopsy sections (p less than 0.001). Fifty-two marrows that were done for staging nonhematological malignancies revealed malignant cells in 21 cases, biopsy sections were positive in all, biopsy imprints were positive in 19 (90%), and aspirate particle smears were positive in 7 (33%). Thirty-six marrows done for staging non-Hodgkin's lymphoma showed malignant cells in 13 cases. Twelve (92%) biopsy sections, three (23%) biopsy imprints, and nine (69%) aspirate particle smears contained lymphoma cells. In conclusion, a satisfactory evaluation of marrow samples for diagnostic studies can be achieved by examination of biopsy sections along with aspirate particle smears or biopsy imprints. Any of the three marrow preparations alone is not sufficient for accurate diagnosis in all cases. The biopsy imprint is an accurate modality for identifying nonhematological tumor metastasis in the bone marrow.     

Persistence of multilineage host haemopoiesis following allogeneic bone marrow transplantation

Summary. The survival of host cells following high-dose cytotoxic therapy and allogeneic marrow transplantation has been established previously, but the identity of these cells has not been elucidated in detail. Four patients who received sex-mismatched marrow have been studied for up to 12 months post-transplant using a simultaneous immuno-phenotyping/fluorescence in situ hybridization technique. The results demonstrate residual host T cells (CD3⁺), B cells (CD22+) and myeloid cells (CDllc+ and CD13+), and additionally cells of progenitor cell phenotype (CD34⁺). The long-term persistence of host haemopoiesis may have major relevance to the post-transplant complications of marrow rejection, graft-versus-host disease, and malignant relapse.     

罗曼鹤鸡骨髓间充质干细胞的分离培养和鉴定

目的建立鸡骨髓间充质干细胞（BMSCs）的分离培养和鉴定体系。方法从1～14天龄罗曼鹤鸡骨髓中分离骨髓细胞,差速贴壁法纯化、扩增BMSCs。倒置显微镜下观察细胞形态特征,MTY法绘制生长曲线,流式细胞仪检测细胞标志物,并分别进行成骨、成脂诱导分化能力检测。结果分离培养的细胞呈成纤维细胞样或长梭形,生长状态良好;CD29阳性表达率为92．10％,CD34阳性表达率仅为0．80％;经成骨诱导分化,出现明显的钙化结节,茜素红染色阳性;经成脂诱导分化,油红O染色阳性,细胞内出现明显的脂质小滴。结论建立了操作简单、高效的鸡BMSCs分离培养和鉴定体系,为鸡BMSCs的进一步研究和应用提供了良好的基础。     

Avascular necrosis of bone after allogeneic bone marrow transplantation: clinical findings, incidence and risk factors

Summary. In the present study we describe the incidence, clinical course, and management of avascular necrosis of bone following allogeneic bone marrow transplantation, and identify risk factors related to its development. All patients developing avascular necrosis of bone after allogeneic bone marrow transplantation between January 1974 and September 1992 were included in the analysis and were studied using the Hôpital Saint Louis Bone Marrow Transplant Database and hospital records. 27/727 allogeneic transplant recipients developed avascular necrosis leading to an 8·1% incidence at 5 years, by product limit estimate, ranging from 5% to 11·2%. Symptoms developed 119–1747 d (median 398 d) after transplantation. In these 27 patients a total of 52 joints were affected (mean 1·92 per patient, range 1–7). The hip joint was most often affected (69% of patients). All patients had joint pain that led to diagnosis by means of standard radiographs with or without the help of technetium-99 scans and/or magnetic resonance imaging. All but three patients received steroid therapy for acute graft-versus-host disease. Among 10 factors tested, three were shown to be significantly linked to an increased risk for developing avascular necrosis by multivariate analysis: male gender (relative risk (RR) 4·2, P= 0·002), age older than 16 (RR = 3·87, P= 0·004), and acute graft-versus-host disease requiring steroid therapy (RR = 6·30, P= 0·0002). 10 patients (37%) required joint replacement within 19 months (range 2–42) following diagnosis of avascular necrosis. In conclusion, avascular necrosis of bone is a frequent late complication of allogeneic bone marrow transplantion causing significant morbidity and requiring replacement surgery in one-third of affected patients. In this 18-year single-centre survey, older age, male gender and steroid therapy given for acute graft-versus-host disease were shown to independently increase the risk of avascular necrosis of bone.     

异种骨复合自体骨髓修复骨缺损的实验研究

目的　观察异种骨复合自体红骨髓对骨缺损的修复能力。方法　选健康纯系 ６ 月龄大耳白兔 ２０ 只,双侧桡骨中段制成 １５～１８ ｃｍ 的骨缺损模型,骨蜡封闭两断端骨髓腔,均植入等长度的异种骨,左侧注射红骨髓作为实验组,右侧不注射作为实验对照组。术后 ２、４、６、８ ｗ 拍 Ｘ 线片,行组织学检查。结果　实验组 ８ ｗ 时, Ｘ 线示骨缺损修复良好、髓腔基本相通,组织学检查见皮质骨形成、骨髓腔相通。而对照组 ８ ｗ 时 Ｘ 线下仍可见未完全吸收的移植骨,光镜示缺损区有残存的坏死骨,纤维组织充填缺损区。结论　异种骨复合自体红骨髓后可明显促进骨融合,提高修复骨缺损的能力     

骨髓移植采髓术麻醉方式的选择

目的：为骨髓移植采髓术寻找一种安全、有效、简便和经济的麻醉方式。方法;对18例行自体骨髓移植术的恶性血液病患者采用局部麻醉方式进行采髓,局麻药为利多卡因,平均用量为93．3mg。结果：平均采髓量862.5ml,术中患者无疼痛感,术后局部有微痛。结论：在局麻下采髓是安全、有效,简便、经济的麻醉方式。     

茶黄素对人骨髓基质干细胞的成骨诱导作用

目的探讨茶黄素对体外培养人骨髓基质干细胞（BMSCs）的成骨诱导作用及其能力。方法将第二代人BMSCs分为茶黄素组、对照组,茶黄素组加入浓度为10 mmol/L茶黄素,对照组不干预。对比观察两组干预后细胞形态,钙结节、碱性磷酸酶（ALP）、Ⅰ型胶原及骨钙素（OCN）免疫组化染色结果,并检测各组4、8、12、16 d各时点ALP、OCN活性。结果茶黄素组钙结节、ALP、Ⅰ型胶原及OCN免疫组化染色均呈阳性,对照组均为阴性;茶黄素组各时点ALP、OCN活性均高于对照组（P均〈0.05）。结论茶黄素可促进人BMSCs向成骨细胞分化。     

冻干骨复合骨髓修复兔桡骨骨缺损的实验研究

目的 比较复合自体红骨髓冻干骨和单纯冻干骨对骨缺损的修复能力。方法 制备兔双侧桡骨骨缺损(1．5cm)模型，实验侧(左侧）植入复合自体红骨髓的冻干骨、对照侧植入单纯冻干骨，术后及术后2、4、6、8w行X线和组织学检查。结果 实验组于术后6w、对照组于术后8w,骨缺损修复良好、髓腔基本相通。结论 复合自体红骨髓的冻干骨修复骨缺损的能力优于单纯冻干骨。