Bone marrow-disseminated tumor cells in patients with carcinoma of the esophagus or cardia

BACKGROUND: The long-term prognosis after surgical therapy for esophageal carcinoma depends on tumor stage and completeness of resection. Similarly to other epithelial tumors, the presence of micro deposits of neoplastic cells in the bone marrow may indicate residual disease and the potential for recurrence. This study assesses the prevalence of bone marrow-disseminated tumor cells in patients undergoing surgical resection for esophageal carcinoma. In addition, we investigated the agreement between immunohistochemical and molecular techniques for the detection of micrometastases in a subgroup of patients. METHODS: Between January 1998 and November 1999, forty-eight patients with adenocarcinoma of the esophagogastric junction (n = 29) or squamous cell carcinoma of the thoracic esophagus (n = 19) and no evidence of overt metastatic disease entered the study. An immunohistochemical assay (capable of detecting 1 carcinoma cell in 7 x 10(5) bone marrow cells) was used to test bone marrow obtained by flushing a resected rib or by needle aspiration either of the iliac crest or of a rib. A polymerase chain reaction (PCR) molecular technique was also used to identify bone marrow and peripheral blood epithelial cells. RESULTS: Cytokeratin-positive cells were found in 79.1% of the bone marrow samples obtained from the rib, and in only 8% of the needle aspirates either from the iliac crest or from a contiguous rib: This difference is probably explained by the improved removal of metastatic cells with the flushing of the rib. Comparable results were obtained at a qualitative level by the PCR technique on bone marrow. In addition, PCR-positive results were found in 3 of 18 peripheral blood samples. There was no association with tumor type, neoadjuvant therapy, or lymph node status. Patients with a pT3 or pT4 tumor showed, at a borderline statistical level, a higher proportion of cytokeratin-positive cells in the flushed rib. CONCLUSIONS: Bone marrow-disseminated tumor cells are present in the resected rib of a high proportion of patients undergoing esophagectomy for carcinoma, and immunohistochemistry seems to be the method of choice for their quantitative assessment. However, the prognostic and therapeutic implications of this finding need further investigation.     

Reconstruction of anterior iliac crest bone graft donor sites: presentation of a surgical technique

This study is a prospective evaluation of the reconstruction of anterior iliac crest bone graft donor sites using a technique developed by the authors. We present the technique and the initial results obtained with its use in 15 patients followed up for a period ranging from 6 to 16 months. Reconstruction of the iliac crest is performed using a rib, which is removed by an anterior approach to the spine. The rib is divided into two segments, which are fitted into the defect created in the iliac crest after removal of the bone graft. Clinical evaluation of the patients found a good cosmetic appearance of the reconstruction site, and the rib segments used showed good radiologic integration. Partial resorption of the segments was observed in two patients, with no effects on the cosmetic result. Received: 3 May 1999 Revised: 13 August 1999 Accepted: 18 August 1999     

Primary lymphoma of bone originating in a rib.

Malignant lymphoma originating in the bone is rare and is now recognized as being an independent clinicopathologic entity known as primary lymphoma of bone. A 60-year-old man complaining of right chest and back pain consulted our hospital for further examination. Chest X-ray and computed tomogram revealed osteolysis and a surrounding soft tissue mass in the sixth right rib. An ultrasonically-guided needle biopsy of the tumor was performed, and histologic examination indicated the dense proliferation of similar-sized atypical cells with nucleoli and an irregular nuclear border. A diagnosis of diffuse, medium-sized non-Hodgkin's lymphoma, B-cell type was made. En block resection of the tumor and chest wall was performed. Macroscopically, the tumor measured 7.5 x 4.8 x 3.0 cm in diameter, and the histologic findings were similar to those of the preoperative needle biopsy. Unfortunately, postoperative treatment with radiation therapy and chemotherapy was ultimately unsuccessful, and a local recurrence and metastatic lesions appeared in the stomach and para-aortic abdominal lymph nodes 7 months after the first symptom appeared. The patient died 3 months later. Surgery was chosen as the initial therapy as it was considered that a rib resection would not result in serious respiratory compromise and the complete resection of the tumor would be superior to radiation therapy for local control. Some authors have reported that the surgical resection of a primary lymphoma of the bone originating in a rib can yield a good prognosis. However, it is a systemic disease and a more effective therapeutic strategy should be developed.     

Unsuspected lymphoma diagnosed with use of biopsy during kyphoplasty

BACKGROUND: Vertebral augmentation procedures are currently widely performed to treat vertebral compression fractures. In selecting appropriate patients for these procedures, it is important to distinguish the pain caused by a fracture from other causes of back pain. The purpose of this study was to determine the frequency of underlying, previously unrecognized malignant tumors in a consecutive series of patients undergoing kyphoplasty to treat vertebral compression fractures. Our hypothesis was that an unsuspected malignant tumor will exist and that a bone-marrow aspiration from the iliac crest would enhance our ability to detect a malignant tumor. METHODS: A prospective histological evaluation of vertebral body biopsy specimens from presumed osteoporotic vertebral compression fractures and a concurrent bone-marrow aspiration from the iliac crest were performed in order to identify latent hematopoietic dyscrasias. Over a four-year period, vertebral body biopsies from 523 vertebral levels as well as iliac crest bone-marrow aspirations were performed in 238 patients. Both specimens were evaluated histologically, and the prevalence of an underlying occult malignant neoplasm was determined. RESULTS: All specimens from the vertebral bodies showed signs of bone-remodeling and/or fracture-healing. However, in three patients, both the bone biopsy specimen and the bone-marrow aspirate showed evidence of B-cell lymphoma. The bone-marrow aspirate did not provide any additional information compared with the vertebral body biopsy specimen, and multiple myeloma was not identified in any patient. CONCLUSIONS: Lymphoma is an uncommon cause of a vertebral compression fracture, but on the basis of our experience in this series, we recommend that vertebral body biopsy specimens be obtained in all patients managed with kyphoplasty and vertebroplasty to rule out an unsuspected malignant tumor. However, we do not recommend the routine use of an additional bone-marrow aspiration from the iliac crest during vertebral augmentation procedures because doing so did not appear to enhance our ability to detect a malignant tumor.     

Editorial Commentary: Bone Marrow Aspirate Concentrate: Time to Harvest Locally?

Cell therapies hold great promise as primary and adjuvant treatments for a range of musculoskeletal conditions. Bone marrow harvested from the iliac crest represents the gold-standard source of progenitor cells with a recognized ability to release trophic factors, modulate local immune environments, and differentiate into multiple musculoskeletal cell types in vitro. Identifying accessible locations that limit donor-site morbidity while increasing efficiency during aspiration of bone marrow is essential. There is increasing evidence to suggest that the number of progenitor cells present in bone marrow aspirated from multiple sites, including the proximal humerus and ilium, is at least equivalent to that from the iliac crest. Because many of these sources lie within the surgical field, the requirement for iliac crest harvest and the risks associated with secondary harvest sites can be mitigated. Although there is a clear need for further studies evaluating the biological attributes and clinical benefit of bone marrow aspirate concentrate in a range of clinical settings, the use of local harvesting sites is likely to reduce morbidity and improve the experience for patients.     

The comparative values of bone marrow aspirate and trephine for obtaining bone scan-targeted metastases from hormone-refractory prostate cancer

Samples of metastatic prostate cancer to bone are difficult to obtain. The aim of this study was to compare the results of bone marrow aspirate and trephine biopsy for obtaining metastatic hormone-refractory prostate cancer (HRPC) samples using previous diagnostic planar 99(m)Tc-HDP bone scans to guide the procedure. All samples taken were for the purposes of research and molecular studies on HRPC. Twenty patients with HRPC had bone marrow aspirate and trephines taken from lesions in the posterior superior iliac spine or sacro-iliac region when shown on diagnostic 99(m)Tc-HDP bone scans. Three patients also underwent plain X-ray, 18F-positron emission tomography bone scan, pelvic MRI scan and 99(m)Tc nanocolloid bone marrow scans. These images were used to assess if the extra imaging information provided, such as three-dimensional localisation of the bone metastases, was of value for target bone metastases. Cancer cells were obtained in 15/20 (75%) cases in which a trephine biopsy was attempted and 0/20 of cases in which a bone marrow aspiration was attempted. The additional information provided by the range of other imaging investigations was of little benefit in obtaining tumour samples, but did suggest why negative biopsies were obtained in some cases after targeting with planar bone scans. We recommend the use of bone marrow trephine biopsy alone, guided by previous diagnostic 99(m)Tc planar bone scan as a practical method to obtain prostate cancer cells from bone metastases.     

Simplified bone marrow biopsy needle for staging of carcinoma of prostate

Bone marrow biopsy and bone marrow aspiration for acid phosphatase determination are simple adjuncts to the staging of carcinoma of the prostate. The Vim-Silverman needle has been modified to a double-sheath needle to obtain a bone marrow biopsy and bone marrow aspirate with a single puncture of the bone.     

The vertebral interbody grafting site’s low concentration in osteogenic progenitors can greatly benefit from addition of iliac crest bone marrow

The ability of bone substitutes to promote bone fusion is contigent upon the presence of osteoinductive factors in the bone environment at the fusion site. Osteoblast progenitor cells are among these environmental osteoinductive factors, and one of the most abundant and available sources of osteoblastic cells is the bone marrow. As far as biological conditions are concerned, the vertebral interbody space appears as a favorable site for fusion, as it is surrounded by spongy bone, theoretically rich in osteogenic cells. This site may, however, not be as rich in osteogenic precursor cells especially at the time of grafting, because decortication of the vertebral end plates during the grafting process may modifiy cell content of the surrounding spongy bone. We tested this hypothesis by comparing the abundance of human osteogenic precursor cells in bone marrow derived from the iliac crest, the vertebral body, and the decorticated intervertebral body space. The number of potential osteoblast progenitors in each site was estimated by counting the alkaline phosphatase–expressing colony-forming units (CFU-AP). The results, however, demonstrate that the vertebral interbody space is actually poorer in osteoprogenitor cells than the iliac crest (P<0.001) and vertebral body (P<0.01), especially at the time of graft implantation. In light of our results, we advocate addition of iliac crest bone marrow aspirate to increase the success rate of vertebral interbody fusion.     

Operative technique of distal radius resection and wrist fusion with iliac crest bone graft

Malignant lesions of distal radius and appropriately selected cases of benign aggressive lesions (giant cell tumor) of distal radius require resection for limb salvage. Post resection, reconstruction of that defect can be accomplished by either arthrodesis or arthroplasty both having their own pros and cons. In cases undergoing arthrodesis as modality of reconstruction, small defects (≤6 cm) can be reconstructed using autologous iliac crest bone graft which results in good cosmetic appearance and functional outcome. We have described in detail, the preoperative planning, surgical steps and rehabilitation of wrist fusion with iliac crest bone grafting post distal radius resection.     

Bone marrow micrometastases in breast cancer patients

BACKGROUND: The status of the axillary nodes has historically been the most important indicator of prognosis in breast cancer patients. However, approximately one third of node-negative patients recur with systemic disease. The detection of bone marrow micrometastases (BMM) may represent additional information in predicting distant recurrence and survival. METHODS: Bone marrow aspiration is obtained from the patient's anterior iliac crest at the time of breast cancer surgery. Cytospins are prepared from this aspirate and stained for polymerase chain reaction (PCR) analysis. RESULTS: Multiple studies have evaluated the clinical implications of BMM. The majority of studies have found a significant correlation between the presence of BMM and decreased survival. The information of bone marrow status may serve as a complement to axillary lymph node status in assessing the prognosis of breast cancer patients. CONCLUSIONS: There is a strong correlation between the presence of bone marrow micrometastases and poorer survival. These results may have an impact upon therapeutic recommendations in breast cancer patients.     

Experimental posterolateral spinal fusion with beta tricalcium phosphate ceramic and bone marrow aspirate composite graft

Background:

Beta tricalcium phosphate is commonly used in metaphyseal defects but its use in posterolateral spinal fusion remains controversial. There are very few published animal studies in which use of beta tricalcium phosphate has been evaluated in the posterolateral lumbar arthrodesis model. Hence we conducted a study to evaluate the potential of composite graft of beta tricalcium phosphate and bone marrow aspirate in comparison to autologous bone graft, when used for posterolateral spinal fusion.

Materials and Methods:

Single level posterolateral lumbar fusion was performed in 40 adult male Indian rabbits, which were assigned randomly into one of the four groups based on graft materials implanted; a) 3 gm beta tricalcium phosphate plus 3 ml bone marrow aspirate (Group I); b) 3 ml bone marrow aspirate alone (Group II); c) 3 gm beta tricalcium phosphate (Group III) and d) 3 gm autologous bone graft (Group IV). Each group had 10 rabbits. Half of the rabbits were sacrificed by injecting Phenobarbitone intraperitoneally after eight weeks and the remaining after 24 weeks, and were evaluated for fusion by X-rays, computed tomography (CT) scans, manual palpation test and histology.

Results:

Beta tricalcium phosphate used with bone marrow aspirate produced best results when compared to other groups (P =.0001). When beta tricalcium phosphate was used alone, fusion rates were better as compared to fusion achieved with autologous iliac crest bone graft though statistically not significant (P =0.07). Autologous bone graft showed signs of new bone formation. However, the rate of new bone formation was comparatively slow.

Conclusion:

Composite graft of beta tricalcium phosphate and bone marrow aspirate can be used as an alternative to autologous iliac crest bone graft.     

Age- and gender-related changes in the cellularity of human bone marrow and the prevalence of osteoblastic progenitors.

Bone marrow harvested by aspiration contains connective tissue progenitor cells which can be induced to express a bone phenotype in vitro. The number of osteoblastic progenitors can be estimated by counting the colony-forming units which express alkaline phosphatase (CFU-APs). This study was undertaken to test the hypothesis that human aging is associated with a significant change in the number or prevalence of osteoblastic progenitors in the bone marrow. Four 2-ml bone marrow aspirates were harvested bilaterally from the anterior iliac crest of 57 patients, 31 men (age 15-83) and 26 women (age 13-79). A mean of 64 million nucleated cells was harvested per aspirate. The mean prevalence of CFU-APs was found to be 55 per million nucleated cells. These data revealed a significant age-related decline in the number of nucleated cells harvested per aspirate for both men and women (P = 0.002). The number of CFU-APs harvested per aspirate also decreased significantly with age for women (P = 0.02), but not for men (P = 0.3). These findings are relevant to the harvest of bone marrow derived connective tissue progenitors for bone grafting and other tissue engineering applications, and may also be relevant to the pathophysiology of age-related bone loss and post-menopausal osteoporosis.     

In vivo evaluation of coralline hydroxyapatite and direct current electrical stimulation in lumbar spinal fusion.

STUDY DESIGN: An animal model of posterolateral intertransverse process lumbar spinal fusion using autologous bone, coralline hydroxyapatite, and/or direct current electrical stimulation. OBJECTIVES: To evaluate the effect of an osteoconductive bone graft substitute and direct-current electrical stimulation on the rate of pseudarthrosis in a rabbit spinal fusion model. SUMMARY OF BACKGROUND DATA: Conventional techniques for the surgical treatment of degenerative conditions in the lumbar spine have a substantial failure rate and associated morbidity. Bone graft substitutes and electrical stimulation are alternative techniques to enhance fusion rates and limit the morbidity associated with posterolateral intertransverse process fusion using autologous iliac crest bone graft. METHODS: Fifty-three adult female New Zealand White rabbits underwent single-level lumbar posterolateral intertransverse process fusion. Animals were assigned to one of four groups using either autologous bone (Group I), coralline hydroxyapatite with autologous bone marrow aspirate (Group II), coralline hydroxyapatite with a 40-microA implantable direct current electrical stimulator and bone marrow aspirate (Group III), or coralline hydroxyapatite with a 100-microA implantable direct current electrical stimulator and bone marrow aspirate (Group IV). Animals were killed at 8 weeks, and fused motion segments were subjected to manual palpation, mechanical testing, and radiographic and histologic analysis to assess the fusion mass. RESULTS: Successful fusion was achieved in 57% (8/14) of animals in Group I, 25% (3/12) in Group II, 50% (6/12) in Group III, and 87% (13/15) in Group IV. Mean stiffness and ultimate load to failure were significantly higher in Group IV than in all other groups (P < 0.05). Histologic analysis demonstrated a qualitative increase in fusion mass in Group IV versus all other groups. CONCLUSIONS: Direct-current electrical stimulation increased fusion rates in a dose-dependent manner in a rabbit spinal fusion model. Coralline hydroxyapatite is an osteoconductive bone graft substitute, and thus requires an osteoinductive stimulus to ensure reliable fusion rates. Furthermore, coralline hydroxyapatite and direct current electrical stimulation can be used together to increase fusion rates in a rabbit spinal fusion model while avoiding the morbidity associated with harvesting iliac crest bone.     

Core decompression with marrow stem cells

Association of core decompression with technique of bone graft is frequent and many different techniques have been described. In this series, grafting was done with autologous bone marrow obtained from the iliac crest of patients operated for hips osteonecrosis. The bone marrow was harvested under general anesthesia. A beveled metal trocar of 6 to 8 cm in length and a bore of 1.5 mm was pushed deep into the cancellous bone. A 10-mL syringe that has been flushed with heparin is used to aspirate the marrow. Once the needle has been inserted to the desired depth, the tip is swept around a full circle in 45° steps, with the bevel pointing in different directions at each step. This procedure is continued until a sufficient quantity of bone marrow has been harvested (150 mL). All the marrow aspirated is discharged into a plastic collection bag containing ACD (acid citrate dextrose) anticoagulant solution. It is then filtered, to remove fat aggregates and clots. The aspirated marrow was reduced in volume by concentration and injected in the femoral head after core decompression with a small trocar. To measure the number of progenitor cells transplanted, we used the fibroblast colony forming unit as an indicator of the stromal cell activity and performed in vitro cultures of the fibroblast progenitor cells. The average volume of bone marrow aspiration was 147 mL ± 12 mL per hip. The number of progenitor cells was average 12.4 ± 3.4 per 10⁶ bone marrow nucleated cells and the number of nucleated cells was estimated to be 16.4 million cells per milliliter of bone marrow.     

Biological implications of tumor cells in blood and bone marrow of pancreatic cancer patients

BACKGROUND: Patients with pancreatic cancer often have tumor recurrence despite curative resection. Cancer cells detected in blood or bone marrow at the time of diagnosis may relate to tumor stage and to prognosis. Recent research emphasis has centered on tumor cells in bone marrow aspirates, but whether these represent early micrometastases or blood-borne cells in transit is unknown. PATIENTS AND METHODS: We developed a specific immunocytochemical assay that evaluated more than 5.3 x 10(6) extracted mononuclear cells per sample of blood and bone marrow and that could identify a single tumor cell in that population. The assay was applied to samples of blood and bone marrow from 105 patients with pancreatic cancer and 66 controls. The prevalence of isolated tumor cells was compared with Union Internationale Contre le Cancer (UICC) stage. A multivariate Cox regression analysis for survival was performed. RESULTS: Pancreatic cancer cells were detected in 26% of blood samples and in 24% of bone marrow specimens. Specificity for cancer was 96%. The prevalence of isolated tumor cells in patients with proven resectable cancer was 9% in blood and 13% in bone marrow. The prevalence increased with UICC tumor stage in blood (P =.04) but not in bone marrow (P =.52) and correlated in blood with resectability (P =.02), progression of disease (P=.08), and peritoneal dissemination (P =.003). While survival correlated significantly with tumor stage (P <.001) and isolated tumor cells in blood correlated with tumor stage, the finding of cancer cells in blood or bone marrow, or both, was not independently associated with survival in patients with pancreatic cancer. CONCLUSIONS: Isolated tumor cells in blood but not bone marrow reflect the stage of growth and spread of pancreatic cancer, particularly in the peritoneal cavity. The findings are consistent with cells in bone marrow aspirates being in transit, not implanted. These disseminated cancer cells may be the consequence, rather than the cause, of progression.     

Detection of hematogenous tumor cell dissemination predicts tumor relapse in patients undergoing surgical resection of colorectal liver metastases

OBJECTIVE: To examine the prognostic significance of disseminated tumor cells in blood and bone marrow of patients undergoing surgical resection of colorectal liver metastases. SUMMARY BACKGROUND DATA: Despite curative hepatic resection of colorectal liver metastases, a high percentage of patients develop tumor recurrence. These recurrences probably originate from disseminated tumor cells released into the circulation before or during surgery. METHODS: Thirty-seven patients with potentially curative (R0) resection of colorectal liver metastases were prospectively enrolled into the study. Preoperative bone marrow samples and preoperative, intraoperative, and postoperative blood samples were examined for disseminated tumor cells by CK20 RT-PCR. RESULTS: Tumor cells were detected in preoperative blood samples in 11 of 37 (30%) patients, in intraoperative blood samples in 17 of 37 (46%) patients, and in postoperative blood samples in 8 of 37 (22%) patients. Four of 25 (16%) patients tested positive for disseminated tumor cells in bone marrow samples. Median follow-up time for all patients was 38 months (range, 10-63 months). Multivariate analysis confirmed tumor cell detection in intraoperative blood (P = 0.009) and in bone marrow samples (P = 0.013) to be independent prognostic factors of tumor relapse. CONCLUSIONS: This is the first study demonstrating that detection of hematogenous tumor cell dissemination during hepatic resection of colorectal cancer metastases predicts tumor relapse. Detection of disseminated tumor cells may help to individualize adjuvant therapy for patients with colorectal liver metastases and to develop surgical strategies to prevent intraoperative hematogenous tumor cell shedding.     

Aspiration of osteoprogenitor cells for augmenting spinal fusion: comparison of progenitor cell concentrations from the vertebral body and iliac crest

BACKGROUND: Successful arthrodesis in challenging clinical scenarios is facilitated when the site is augmented with autograft bone. The iliac crest has long been the preferred source of autograft material, but graft harvest is associated with frequent complications and pain. Connective tissue progenitor cells aspirated from the iliac crest and concentrated with allograft matrix and demineralized bone matrix provide a promising alternative to traditional autograft harvest. The vertebral body, an even larger reservoir of myeloproliferative cells, should provide progenitor cell concentrations similar to those of the iliac crest. METHODS: Twenty-one adults (eleven men and ten women with a mean age of 59 +/- 14 years) undergoing posterior lumbar arthrodesis and pedicle screw instrumentation underwent transpedicular aspiration of connective tissue progenitor cells. Aspirates were obtained from two depths within the vertebral body and were quantified relative to matched, bilateral aspirates from the iliac crest that were obtained from the same patient at the same time. Histochemical analysis was used to determine the prevalence of vertebral progenitor cells relative to the depth of aspiration, the vertebral level, age, and gender, as compared with the iliac crest standard. The cell count, progenitor cell concentration (cells/cc marrow), and progenitor cell prevalence (cells/million cells) were calculated. RESULTS: Aspirates of vertebral marrow demonstrated comparable or greater concentrations of progenitor cells compared with matched controls from the iliac crest. Progenitor cell concentrations were consistently higher than matched controls from the iliac crest (p = 0.05). The concentration of osteogenic progenitor cells was, on the average, 71% higher in the vertebral aspirates than in the paired iliac crest samples (p = 0.05). With the numbers available, there were no significant differences relative to vertebral body level, the side aspirated, the depth of aspiration, or gender. An age-related decline in cellularity was suggested for the iliac crest aspirates. CONCLUSIONS: The vertebral body is a suitable site for aspiration of bone marrow for graft augmentation during spinal arthrodesis.     

Iliac crest biopsy: Representativity for the amount of mineralized bone

The aim of the study was to evaluate the representativity of iliac crest biopsy for the amount of mineralized trabecular and cortical bone in the skeleton. The following data were obtained on bone from 14 necropsies: right sided iliac crest biopsy, lumbar spine biopsy, dry fat free weight of lumbar spine, bone mineral density (BMD) in the lumbar spine and dry fat free weight of cortical and trabecular bone from the left distal forearm.

The amount of mineralized cortical and trabecular bone from various sites was compared by linear regression analysis. The results confirm iliac crest biopsy as a good predictor of the amount of trabecular bone, but not of cortical bone. Furthermore, iliac crest biopsy is a better estimate of the amount of trabecular bone in the lumbar spine than spinal BMD.