Background Bone Marrow Research Articles

Diagnostic outcomes of bone marrow aspirate and trephine biopsies performed at a hospital in KwaZulu-Natal, South Africa.

BackgroundBone marrow aspiration and trephine biopsy (BMAT) are widely performed in adults to evaluate haematological and malignant conditions. However, the diagnostic yield from the procedure in unselected patients in the South African public sector has not previously been described.ObjectivesWe identified the main indications and most common diagnoses encountered for BMAT and described the demographic and blood profiles of patients, including HIV-positive patients, who had undergone the procedure at a tertiary hospital in KwaZulu-Natal.MethodsWe retrospectively reviewed laboratory data from January 2016 to December 2016 for all patients aged ≥ 13 years who underwent the procedure and stratified findings by demographic data.ResultsAmong 120 BMAT biopsies studied, 80 (67%) cases were performed to evaluate suspected malignancy and a further 40 (33%) cases for non-malignant indications. The main indications for bone marrow examination were: cytopenias 38 (32%), lymphoma 35 (29%), leukaemia 21 (18%), and multiple myeloma 17 (14%). BMAT results revealed that 60 cases (50%) were malignant in origin, 30 cases (25%) were non-malignant and 30 cases (25%) were classified as normal. The common diagnoses were: leukaemia, 24 (20%); multiple myeloma, 16 (13%) and lymphoma, 13 (11%). Cases aged ≥ 50 years were more likely to have a malignant diagnosis (odds ratio: 5.8 (95% confidence interval: 2.2–17.1) p-value < 0.001).ConclusionThe diagnostic yield of BMAT was high, with significant abnormalities detected in three quarters of cases. Haematological malignancy was the more common diagnosis. Increasing age was associated with an increase in reporting of haematology malignancy.

Phase I and registry study of autologous bone marrow concentrate evaluated in PDE5 inhibitor refractory erectile dysfunction

BackgroundBone marrow mononuclear cells have been successfully utilized for numerous regenerative purposes. In the current study, patients suffering from erectile dysfunction (ED) unresponsive to phosphodiesterase 5 inhibitors were administered autologous bone marrow concentrate delivered intracavernously utilizing a point of care FDA cleared medical device.MethodsA total of 40 patients were treated in the primary trial and 100 in the clinical registry, with the longest follow up of 12 months.ResultsMinimal treatment associated adverse effects where observed related to short term bruising at the site of harvest or injection. No long-term adverse events were noted related to the intervention. Mean improvements in IIEF-5 score were 2 in the Caverstem 1.0 low dose group, 3 in the high dose Caverstem 1.0 group and 9 in the Caverstem 2.0 group. Furthermore, improvements peaked by 3 months and maintained at 6 months follow-up.ConclusionThese data support the safety and efficacy of point of care, minimally to non-manipulated, non-expanded bone marrow concentrate for the treatment of ED.Trial registration Funded by Creative Medical Health, Inc.; Clinicaltrials.gov number: NCT03699943; https://clinicaltrials.gov/ct2/show/NCT03699943?term=caverstem&rank=1; initially registered December 12, 2015.

Decoding Bone Marrow Fibrosis in Myelodysplastic Syndromes

BackgroundBone marrow fibrosis (BMF), a poor prognostic factor in myelodysplastic syndromes (MDS), in the context of new risk stratifications of MDS has not been fully explored. We examined the relationship between BMF in MDS and survival outcomes, and explored the landscape of somatic gene mutations in the setting of BMF. Patients and MethodsWe retrospectively evaluated 2624 MDS patients for BMF who were divided into 2 groups: grade 0-2 BMF (96%) and severe/grade 3 BMF (4%) based on analysis presented. Commonly MDS tested acquired somatic mutations were also compared between those 2 groups of patients with available next-generation sequencing data. ResultsOnly grade 3 BMF was associated with worse overall survival independent from the Revised International Prognostic Scoring System (IPSS-R) (hazard ratio = 1.6; 95% confidence interval, 1.2-1.9; P < .005). More patients with severe BMF were classified as MDS-EB1 and -EB2 by the World Health Organization 2016 classification, a higher-risk International Prognostic Scoring System score, and a high/very high IPSS-R risk category than patients with grade 0-2 BMF. A complex karyotype, higher bone marrow myeloblasts, lower platelets, and higher rate of elevated lactate dehydrogenase were observed more often in patients with severe BMF. No differences in response to hypomethylating agents or lenalidomide were observed. Among somatic gene mutations tested in MDS, TP53 mutation and SETBP1 were more frequent in patients with grade 3 BMF. ConclusionThe presence of grade 3 BMF is associated with reduced overall survival independent from IPSS-R; however, BMF grade did not affect response to hypomethylating agent or lenalidomide treatment. TP53 and SETBP1 mutations occurred with greater frequency among patients with severe fibrosis.

IGF-1 enhances BMSC viability, migration, and anti-apoptosis in myocardial infarction via secreted frizzled-related protein 2 pathway

BackgroundBone marrow mesenchymal stem cell (BMSC) transplantation represents a promising therapeutic strategy for ischemic heart disease. However, its effects are hampered by the poor viability of transplanted cells and the hostile microenvironment of the ischemic region. Insulin-like growth factor-1 (IGF-1) is an important paracrine growth factor of BMSC and plays an important role in the properties of BMSC. Here, we investigated whether overexpressing IGF-1 could enhance the BMSC viability, migration, anti-apoptosis, and protective effects of cardiomyocytes, and explore the underlying mechanisms’ focus on the role of the AKT/secreted frizzled-related protein 2 (SFRP2)/β-catenin pathway.MethodsWe constructed BMSCs overexpressing insulin-like growth factor-1 (BMSCs-IGF-1) or empty vector (BMSCs-NC) using lentivirus, and evaluated cell survival, proliferation, and migration under normoxic and hypoxic conditions. Co-culture of rat cardiomyoblasts with BMSCs was performed to explore the paracrine effect of BMSCs-IGF-1 for rescuing cardiomyoblasts under hypoxia. Transplantation of BMSCs in acute myocardial infarction rats was used to explore the effect of BMSCs-IGF-1 therapy.ResultsBMSCs-IGF-1 exhibited a higher cell proliferation rate, migration capacity, and stemness, and were more resistant to apoptosis under hypoxia. Overexpression of IGF-1 upregulated the expression of total and nuclear β-catenin via the AKT-secreted frizzled-related protein 2 (SFRP2) pathway, which enhanced cell survival. Inhibition of AKT or SFRP2 knockdown by siRNA significantly antagonized the effect of IGF-1 and decreased the expression of β-catenin. The expression of β-catenin target genes, including cyclin D1 and c-Myc, were accordingly decreased. Moreover, BMSCs-IGF-1 could rescue cardiomyoblasts from hypoxia-induced apoptosis and preserve cell viability under hypoxia. Transplantation of BMSCs-IGF-1 into myocardial infarction rats greatly reduced infarct volume than BMSCs-NC, with significantly greater expression of SFRP2 and β-catenin.ConclusionsThese results suggest that in BMSCs overexpressing IGF-1, SFRP2 is an important mediator for the enhancement of stem cell viability via activating, rather than antagonizing, the Wnt/β-catenin pathway.

Role of diffusion-weighted magnetic resonance imaging in evaluation of bone marrow edema of the knee

Background Bone marrow edema (BME) has not been recognized as a significant pain generator or as a primary contributor to disease until recently. BME is characterized by a nonspecific pattern of ill-defined high signal intensity on short-tau inversion recovery and T2-weighted MRI and low signal intensity on T1-weighted images. Objective To assess the value of diffusion-weighted MRI in the diagnosis and evaluation of BME at the knee region. Patients and methods This study was conducted on 130 patients referred with clinical suspicion of knee edema with or without history of trauma. They were divided into group ‘A,’ with lesions less than 15 mm, and group ‘B,’ with lesions greater than 15 mm. Their ages ranged between 9 and 75 years, with a mean age of 42 years. There were 43 females and 87 males. Results Between groups A and B, we found that bone marrow lesions (BMLs) were approximately two times larger on apparent diffusion coefficient (ADC) maps than on fat-saturated (FS) proton density (PD)-weighted turbo spin echo images. The most important value of adding the ADC-derived maps in routine MRI examination of the knee is to show additional BMLs that only can be detected at the ADC maps but could not be detected at the FS PD. Approximately 27 more lesions in 27 patients and one patient with two more lesions were detected. These nonconcordant lesions accounted totally for 29 more BMLs. Conclusion MRI is the mainstay of diagnosis, staging, and management plan of knee traumatic and nontraumatic disorders. ADC maps are more sensitive than corresponding FS PD-weighted turbo spin echo images allowing for detection of significantly more and larger BMLs. In addition, they represent a valuable add-on in knee imaging protocols.

Analgesic efficacy of intramuscular ketamine/ketorolac versus fentanyl/ketorolac for children undergoing bone marrow biopsy and aspiration

Background Bone marrow aspiration and/or biopsy (BMAB) in children with hematological disorders is a painful procedure, is required to be repeated at regular intervals, and is under researched. Objective To compare intramuscular administration of fentanyl/ketorolac (F) versus ketamine/ketorolac (K) as an analgesic regimen in pediatric population undergoing BMAB under general anesthesia. A total of 80 children underwent BMAB under general anesthesia, who were randomized into two equal groups: group K received ketamine 0.5 mg/kg/ketorolac 1 mg/kg and group F received fentanyl 1.5 μg/kg/ketorolac 1 mg/kg. The study drugs are injected intramuscularly after induction of general anesthesia. Result No statistically significant difference was found between groups regarding patient characteristics, duration of procedure or duration of anesthesia, pain score, time to first analgesic request, emergence-agitation score, or incidence of vomiting. There was a significant decrease in heart rate, systolic blood pressure, and respiratory rate in both groups at 15 and 20 min compared with basal values. Conclusion Ketamine 0.5 mg/kg+ketorolac 1 mg/kg and fentanyl 1.5 μg/kg+ketorolac 1 mg/kg administered intramuscular are similarly effective analgesic regimens in pediatric population undergoing BMAB under general anesthesia without adverse effects.

Effectiveness of ketamine and propofol (ketofol) in 1:2 versus 1:3 combinations for procedural sedation and analgesia in pediatric patients undergoing bone marrow aspiration and / or biopsy: A prospective cohort study

BackgroundBone marrow aspiration (BMA) and Biopsy is a painful procedure done in children with hematologic disorders. Ketamine and Propofol mixture is widely used for its combined effect of amnesia and analgesia as well as hemodynamic stability and decreased post procedural adverse events. Smooth analgesia and anesthesia is mandatory in such like pediatrics procedures. ObjectiveTo compare the effectiveness of ketofol in 1:2 versus 1:3 combinations for Procedural sedation and analgesia in children undergoing Bone marrow aspiration and Biopsy at Tikur Anbessa Specialized Hospital from December 2019–March 2020. MethodsThis prospective cohort study recruits 128 American Society of Anesthesiologist class I and II, pediatric patients age between 1–12 years undergoing for BMA and Biopsy with ketofol 1:2 or 1:3 combination were included. Hemodynamic, respiratory and RSS (Ramsay sedation score) variables were noted and recorded every 10 min during the procedure. Normality of the data was checked using Kolmogorov-smirnov test and analyzed using student t-test for normally distributed data and chi-square test for categorical data. Non-parametric data was analyzed using Mann –Whitney U test with 95% CI and p-value less than 0.05 is considered as statistically significant. Resultsketofol 1:2 group (n = 64) compared with ketofol 1:3 group (n = 64) had similar sedation level assessed by RSS, hemodynamic and respiratory outcome, as well as general postoperative adverse events profile, but the total intraoperative analgesia consumption was significantly higher in ketofol 1:3 group (29.7%) when compared to ketofol 1:2 group (7.8%) with p = 0.002. ConclusionKetofol 1:3 group shows the need for additional analgesia in this combination. Whereas ketofol 1:2 combination for pediatrics undergoing BMA and Biopsy has decreased intraoperative analgesia requirement.

Variation of the apparent diffusion coefficient of skull bone marrow by age group, pubertal status, and gender in a pediatric population.

Bone marrow composition varies with stage of development. To assess differences in apparent diffusion coefficient (ADC) derived from clivus bone marrow in healthy children by age, pubertal status, and gender as a benchmark when monitoring local and systemic treatment-induced effects. Non-oncological pediatric patients (30 pre-pubertal [15 girls, 15 boys] and 30 post-pubertal [15 girls, 15 boys]) with previous normal magnetic resonance imaging (MRI) of the brain including diffusion-weighted magnetic resonance imaging (DW-MRI; 1.5-T Philips Achieva-Ingenia, b-values 0 and 1000s/mm2) were studied. A 4-6 mm diameter region of interest (ROI), drawn within the clivus on two or three DW-MRI slices, yielded mean and centile ADC values. Pubertal status was recognized from imaging appearances of the pituitary gland and from fusion of the spheno-occipital synchondrosis. Correlations between ADC and age were assessed (Pearson's coefficient). Mann-Whitney U tests compared ADC by age, pubertal status, and gender. Age and ADC were significantly negatively correlated (median ADC r=-0.48, mean ADC r=-0.42, P=0.0001 and 0.0008, respectively) which held true when divided by gender. Mean and median ADC differed significantly before and after puberty for the whole population (P=0.0001 and 0.0001, respectively). There was a left shift of the ADC histogram after puberty with significant differences in centile values. ADC differences before and after puberty remained when divided by gender (girls: P=0.04 and 0.009, respectively; boys: P=0.005 and 0.0002, respectively). ADC of clivus bone marrow correlates with age in children. ADC decreases significantly after puberty, likely due to replacement of hypercellular marrow with fat. There are no gender-related differences in clivus bone-marrow ADC before or after puberty.

Heat shock pretreatment improves mesenchymal stem cell viability by heat shock proteins and autophagy to prevent cisplatin-induced granulosa cell apoptosis

BackgroundBone marrow mesenchymal stem cells (BMSCs) can partially repair chemotherapy-induced ovarian damage. However, low survival rate after transplantation hampers the therapeutic efficiency of BMSCs. Heat shock pretreatment (HSP) effectively improves the cell survival. This study attempted to investigate the mechanisms of HSP on BMSCs survival and the effects of heat shock-pretreated BMSCs (HS-MSCs) on cisplatin-induced granulosa cell (GC) apoptosis.MethodsBMSCs were isolated, cultured, and identified. After receiving HSP for different duration times in a 42 °C water bath, the apoptotic rates of BMSCs were detected by Annexin V-FITC/PI to determine the optimal condition of HSP. Cisplatin was added to the medium of HS-MSCs to simulate chemotherapy environment. The proliferative curve, apoptotic rate, and viability of HS-MSCs were determined by CCK-8, Annexin V-FITC/PI, and Hoechst33342/PI respectively to explore the alteration of biological characteristics. The levels of heat shock protein 70 and 90 (HSP70 and HSP90) and the expressions of autophagy-related markers (Beclin1 and LC3B) were detected by Western blot. In addition, the autophagosomes were observed by transmission electronic microscopy to discuss the possible mechanisms. The GCs were isolated, cultured, and identified. The HS-MSCs were co-cultured with GCs before and after the addition of cisplatin. Then, the apoptotic rate and viability of GCs were detected to investigate the therapeutic and preventive effects of HS-MSCs on GC apoptosis.ResultsAfter receiving HSP at 42 °C for 1 h, BMSCs represented the lowest apoptotic rate. After the addition of cisplatin, the apoptotic rate of HS-MSCs (11.94% ± 0.63%) was lower than that of BMSCs (14.30% ± 0.80%) and the percentage of HS-MSCs expressing bright blue/dull red fluorescence was lower than that of BMSCs. The expression of HSP70 and HSP90 increased, while the number of autophagosomes, the expression of Beclin1, and the LC3BII/LC3BI ratio decreased in HS-MSCs. The apoptotic rates of GCs co-cultured with HS-MSCs before and after the addition of cisplatin were 39.88% ± 1.65% and 36.72% ± 0.96%, both lower than those of cisplatin-induced GCs (53.81% ± 1.89%).ConclusionHSP can alleviate the apoptosis and improve the survival of BMSCs under chemotherapy environment. The mechanism may be associated with the elevated expression of HSP70 and HSP90 and the attenuation of autophagy. Moreover, HS-MSCs have both therapeutic and preventive effects on cisplatin-induced GC apoptosis.

The mechanism of miR-889 regulates osteogenesis in human bone marrow mesenchymal stem cells

BackgroundBone marrow mesenchymal stem cells (BMMSCs) can be used for bone regeneration in the specified condition. Osteogenic differentiation of BMMSCs is controlled by microRNAs (miRNAs) and other factors. This study was aimed to identify the role and mechanism of miR-889 in regulating the osteogenic differentiation of BMMSCs.MethodsOsteoporosis patients and normal control bone tissues were collected and used PCR techniques to identify the change of miR-889 and WNT7A. Moreover, the dynamic change of miR-889 and WNT7A during osteogenic differentiation of BMMSCs was also measured. Bioinformatic analysis was performed to identify the target genes and potential pathways of miR-889. Then, we constructed miR-889 mimic and inhibitor, ALP staining, ARS, osteoblastic-related protein, and Wnt β-catenin signaling pathway-related protein were also measured. WNT7A siRNA was also used to verify the function of miR-889.ResultsIn the present study, we showed that miR-889 expression was upregulated in osteoporosis patients than healthy control. However, the miR-889 expression was downregulated during osteogenic differentiation. Bioinformatics analysis found that miR-889 targets 666 genes and mainly through Wnt β-catenin signaling pathway. Administrated miR-889 mimic, the ALP activity, and calcium deposition were decreased than the control group, while miR-889 inhibitor shown the opposite trend. And miR-889 could bind the 3′UTR of WNT7A. We further used WNT7A siRNA to explore the function of miR-889, and the results revealed that co-cultured with miR-889 inhibitor and WNT7A siRNA was associated with a reduction of ALP activity and calcium deposition and osteoblastic-related proteins than miR-889 inhibitor alone.ConclusionOur results revealed that miR-889 plays a negative role in inducing osteogenic differentiation of BMSCs through Wnt β-catenin signaling pathway.

Next Generation Sequencing May be Helpful in Designing Novel Combinations Among Heavily Pretreated Myeloma Patients, Refractory to All Available Approved Anti-Myeloma Drugs

Introduction: Multiple genetic alterations that occur at diagnosis or relapse are not only prognostic characteristics of multiple myeloma (MM) but also provide evidence for clonal evolution. Uncovering and dissecting true driver events in MM might provide rational for new potential targets and therapeutic approaches. However, whereas genetic diagnostics in MM namely FISH and gene expression profiling are well-established prognostic tools, individual mutation profiling has not yet been adopted for this purpose. Herein, we aimed to analyze the Next Generation Sequencing (NGS) platform results investigating mutational profiling of patients with relapsed and refractory MM (RRMM). Also, the clinical results of those who had a targetable mutation and were treated "off label" will be presented. Material and Methods: A total of 14 consecutive patients with MM referred to our center between November 2018 and May 2019 were studied. Plasma cells were isolated from bone marrow samples using Selection Kit microbead specific for EasySepTM Human CD138 marker (StemCell Technologies). DNA extracted form magnetic bead enriched cells, bone marrow aspiration smears for bone marrow involved, from FFPE tissue samples for extramedullary-involved cases. NGS method was performed on llumina Miseq platform (USA) by using QIAseq targeted DNA panel (12)- Human myeloid neoplasm panel, covers all exons and exon-intron junctions of 141 target genes. For the data analysis QCI Analyze Universal 1,5.0 was performed. The PCL analysis was performed on CD138 and Ki67 double immune stained paraffin sections, and the quantification was done by using 3DHistech digital pathology platform. Results: We obtained 16 samples of DNA from 12 heavily pretreated and two newly diagnosed myeloma patients. Female/male: 5/9 with a median age of 57 years (range, 39-87) patients had received a median four lines (range; 1-13) of treatment. Out of a panel of 141 genes, 59 mutations in 26 genes were detected (Figure-1). Among these recurrent genomic abnormalities, concomitant missense protein coding alterations were detected in all patients. The PTEN mutation was the most frequently detected, followed by mutations of RAS/MAPK pathway genes. The hotspots of mutation in KRAS codon 61 and NRAS included codons 61 and 13 as well as codon 600 in BRAF. In addition, we detected novel ie myeloproliferative and myelodysplasia associated mutations previously un-described in myeloma. A diverse range of recurrent gains and losses were detected in our cohort. Two patients at diagnosis also carried mutations of PTEN and KRAS. Based on these results three patients were able to obtain off-label approval for treatment with Everolimus (for PTEN) (Patient-1) or Trametinib (for KRAS) (Patient 5 &amp; 6) in combination with Pomalidomide (EvoPomDex) w/wo Daratumumab or Tra-PomDex. Patient-1 had extensive extramedullary disease (EMD) in the skin, which responded completely to Dara-EvoPomDex combination. Complete disappearance of initial lesions (presented in Figure-2) and VGPR duration was only two months. Subsequent refractoriness and appearance of new lesions lead to death of the patient, one year from the initiation of EMD. Patient-6, also presented with EMD, was treated with TraPomDex as the seventh treatment line. TraPomDex treatment was well tolerated, the most significant adverse event diarrhea, infections and pancytopenia. Her biochemical response was a transient VGPR, which was lost during interruption of treatment due to infection. She also died four months after initiation of TraPomDex. Patient-5, plasma cell leukemia, has been on Tra-PomDex for a month and his response is PR yet. Conclusion: The detection of mutations can improve our ability to treat multiple refractory patients who have ran out of all therapeutic options. Though the responses observed among such very heavily pretreated patients are not durable, they are highly promising. Also, of additional importance is detection of age-related cumulative mutations belonging to background bone marrow precursors. Detection of sub-clonal mutations is very helpful in depth analysis of clonal response to treatment and clonal evolution. In the coming years, the identification of actionable mutations in myeloma opens the way for targeted therapy. Acknowledgement:This study is supported by Ankara University Research Grants (Project: 14A0230003) and Turkish Academy of Sciences. Disclosures Ozcan: Takeda: Honoraria, Other: Travel support, Research Funding; Bayer: Research Funding; AbbVie: Other: Travel support, Research Funding; Sanofi: Other: Travel support; Abdi Ibrahim: Other: Travel support; Celgene Corporation: Other: Travel support, Research Funding; Janssen: Other: Travel support, Research Funding; Archigen: Research Funding; Roche: Other: Travel support, Research Funding; Jazz: Other: Travel support; MSD: Research Funding; Novartis: Research Funding; Amgen: Honoraria, Other: Travel support; BMS: Other: Travel support. Beksac:Takeda: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene: Speakers Bureau; Janssen: Research Funding, Speakers Bureau; Amgen: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. OffLabel Disclosure: trametinib and everolimus for myeloma patients

The MP0250-CP201 Mirror Study: A Phase 2 Study Update of MP0250 Plus Bortezomib and Dexamethasone in Relapse/Refractory Multiple Myeloma (RRMM) Patients Previously Exposed to Proteasome Inhibitors and Immunomodulatory Drugs

The MP0250-CP201 Mirror Study: A Phase 2 Study Update of MP0250 Plus Bortezomib and Dexamethasone in Relapse/Refractory Multiple Myeloma (RRMM) Patients Previously Exposed to Proteasome Inhibitors and Immunomodulatory Drugs

428P - A pilot study on comparative efficacy of tramadol or eutectic mixture of local anaesthetics (prilocaine plus lignocaine) in preventing bone marrow aspiration associated pain

BackgroundBone marrow aspiration although being a common procedure is associated with significant pain and its reduction remains an unmet need. We evaluated the use of tramadol and eutectic mixture of local anaesthetics (prilocaine plus lignocaine) (EMLA) for reducing the severity of pain. MethodsIn this pilot study, we compared the addition of either Tramadol 50 mg per oral (Arm T) or EMLA local application (Arm E) or no added intervention (Arm L) in addition to local infiltration with lignocaine 2% before bone marrow aspiration in adults suspected / confirmed with malignancy. Both, tramadol and EMLA were administered one hour prior to the procedure. Primary end point was reduction in pain intensity with these interventions compared to local infiltration alone. Pain was assessed using numerical FACES pain scale ( 0-10), a visual analogue scale.Secondary end points were to see the effect on pre procedure apprehension and to find out other factors associated with increased pain associated with the procedure. ResultsA total of 225 patients were included in the study, 75 each in tramadol(T), EMLA(E) and only lignocaine local infiltration(L) arms respectively. The mean pain intensity on the visual scale was significantly lower in tramadol arm ( T,E,L - 3.3, 4.5, 4.7 respeactively)(p < 0.05). There was significant reduction in percentage of patients who experienced moderate / severe pain (4 or more) in tramadol arm (T,E,L - 47%,76%,84% respectively)(p < 0.05). Duration of procedure > 10 minutes and age > 50 were positively correlated with more pain. Leukemia patients had significantly more pain compared to lymphoma and other solid malignancies. Only 4 patients had sedation/dizziness in tramadol arm compared to 2 each in the other two arms. No significant effect on pre procedure apprehension was noted in any of the arms. ConclusionsTramadol appears to have a preventive effect on bone marrow aspiration associated pain and appears to be well tolerated. Whereas EMLA was not associated with such effect. Larger studies may be done to ascertain the same. Legal entity responsible for the studyDepartment of medical oncology, Kidwai cancer institute. FundingHas not received any funding. DisclosureAll authors have declared no conflicts of interest.

Combinatorial treatment of acute myocardial infarction using stem cells and their derived exosomes resulted in improved heart performance

BackgroundBone marrow mesenchymal stem cells (MSCs) are among the most common cell types to be used and studied for cardiac regeneration. Low survival rate and difficult retention of delivered MSCs in infarcted heart remain as major challenges in the field. Co-delivery of stem cell-derived exosomes (Exo) is expected to improve the recruitment and survival of transplanted MSCs.MethodsExo was isolated from MSCs and delivered to an acute myocardial infarction (AMI) rat heart through intramyocardial injection with or without intravenous infusion of atrovastatin-pretreated MSCs on day 1, day 3, or day 7 after infarction. Echocardiography was performed to evaluate cardiac function. Histological analysis and ELISA test were performed to assess angiogenesis, SDF-1, and inflammatory factor expression in the infarct border zone. The anti-apoptosis effect of Exo on MSCs was evaluated using flow cytometry and Hoechst 33342 staining assay.ResultsWe found that intramyocardial delivery of Exo followed by MSC transplantation (in brief, Exo+MSC treatment) into MI hearts further improved cardiac function, reduced infarct size, and increased neovascularization when compared to controls treated with Exo or MSCs alone. Of note, comparing the three co-transplanting groups, intramyocardially injecting Exo 30 min after AMI combined with MSCs transplantation at day 3 after AMI achieved the highest improvement in heart function. The observed enhanced heart function is likely due to an improved microenvironment via Exo injection, which is exemplified as reduced inflammatory responses and better MSC recruitment and retention. Furthermore, we demonstrated that pre-transplantation injection of Exo enhanced survival of MSCs and reduced their apoptosis both in vitro and in vivo.ConclusionsCombinatorial delivery of exosomes and stem cells in a sequential manner effectively reduces scar size and restores heart function after AMI. This approach may represent as an alternative promising strategy for stem cell-based heart repair and therapy.

1097P - Is bone marrow examination indispensable in chronic myeloid leukemia at diagnosis?

BackgroundBone marrow aspiration and biopsy at presentation is important to demonstrate the phase of the disease, to provide material for cytogenetic studies and to demonstrate the myelofibrosis, which have prognostic and therapeutic implications. In the current era of molecular diagnostics, the role of bone marrow examination is questionable. The present study was designed to identify the role of bone marrow examination on phase migration, karyotyping and myelofibrosis. MethodsData of newly diagnosed CML patients seen between January 2013 and June 2018 was retrospectively analysed. CML was confirmed by demonstration of BCR-ABL fusion gene by PCR or FISH. Patients were stratified into chronic phase (CP), accelerated phase (AP) and blast phase (BP) according to WHO diagnostic criteria (2016). ResultsOf 749 patients, 541 patients had evaluable data. The male female ratio was 1.52:1 The median age at presentation was 37 years (range, 9-80 years). According to EUTOS risk stratification, 406 (75.1%) patients were low risk and 135(24.9%) were high risk. Based on bone marrow blasts criterion, phase migration occurred in 16 (2.9%). Of these 16 patients, phase migration from CP to AP, CP to BP and AP to BP were seen in 8 (50%), 4(25%) and 4 (25%) patients, respectively. Data on karyotyping was available in 195 (36%) patients. Of 195 patients, additional cytogenetic abnormalities were detected in 30 (15.3%) which includes, major route abnormalities in 11 (5.6%) patients. Of these 11 patients, 6 patients were in CP, 4 patients were in AP and 1 patient in BP. Myelofibrosis was demonstrated in 264 (48.7%) patients of which grade 1, grade 2, grade 3 and grade 4 were seen in 72 (13.3%), 100 (18.5%), 70 (12.9%); 20 (3.7%) patients respectively. ConclusionsBone marrow examination at presentation in CML patients results in phase migration, provides adequate material for cytogenetic studies and demonstrates presence of myelofibrosis. Hence, bone marrow examination is indispensable in newly diagnosed CML patients. Legal entity responsible for the studyNizam’s Institute of Medical Sciences. FundingHas not received any funding. DisclosureAll authors have declared no conflicts of interest.

Intramyocardial Bone Marrow Stem Cells in Patients Undergoing Cardiac Surgical Revascularization

Bone marrow stromal or stem cells (BMSCs) remain a promising potential therapy for ischemic cardiomyopathy. The primary objective of this study was to evaluate the safety and feasibility of direct intramyocardial injection of autologous BMSCs in patients undergoing transmyocardial revascularization (TMR) or coronary artery bypass graft surgery (CABG). A phase I trial was conducted on adult patients who had ischemic heart disease with depressed left ventricular ejection fraction and who were scheduled to undergo TMR or CABG. Autologous BMSCs were expanded for 3 weeks before the scheduled surgery. After completion of surgical revascularization, BMSCs were directly injected into ischemic myocardium. Safety and feasibility of therapy were assessed. Cardiac functional status and changes in quality of life were evaluated at 1 year. A total of 14 patients underwent simultaneous BMSC and surgical revascularization therapy (TMR+BMSCs= 10; CABG+BMSCs= 4). BMSCs were successfully expanded, and no significant complications occurred as a result of the procedure. Regional contractility in the cell-treated areas demonstrated improvement at 12 months compared with baseline (TMR+BMSCs Δ strain:-4.6% ± 2.1%; P=.02; CABG+MSCs Δ strain:-4.2% ± 6.0%; P= .30). Quality of life was enhanced, with substantial reduction in angina scores at 1 year after treatment (TMR+BMSCs: 1.3 ± 1.2; CABG+MSCs: 1.0 ± 1.4). In this phase I trial, direct intramyocardial injection of autologous BMSCs in conjunction with TMR or CABG was technically feasible and could be performed safely. Preliminary results demonstrate improved cardiac function and quality of life in patients at 1 year after treatment.

Effect of Teaching Program on Nurs's Performance Regarding Bone Marrow Transplantation

Background Bone marrow transplantation is an advanced therapeutic intervention that is required for a number of malignant and nonmalignant medical conditions, often for critically ill patients. the nurses needed to be provided with knowledge and practice to give high quality of care for patient. Aim of the study: was to evaluate The effect of teaching program on nurses`performance regarding bone marrow transplantation. Study Design: Quasi-experimental pre-post test research design was utilized.. Setting The study was conducted in bone marrow transplantation unit and other critical care unit at south Egypt cancer institute at Assiut University hospital Sample: All nurses (30) from above mentioned setting. Tools: An interview questionnaire for the nurse, Tool 1 knowledge assessment tool will be utilized, Tool 2. Observational chicklist for practise assessment tool will be utilized Results nurses knowledge was significant improved from 18.47﴾to﴿32.77 respectively and nurses practice was improved from 48.47 to96.5 after the implementation of the program. Conclusion: There were highly significant improvement in knowledge and practice after implementation of program about bone marrow transplantation. Recommendation; Continues nursing education and training programs at critical care units should be organized within south Egypt cancer institute

Selective local irradiation improves islet engraftment and survival in intra–bone marrow islet transplantation

BackgroundBone marrow (BM) is as an alternative site for islet transplantation, but it is not an immunoprotected microenvironment and allogeneic islets are rejected. However, the BM, for its structure and anatomic position, offers the possibility to modulate microenvironment by local interventions. We here investigate whether local irradiation is able to improve islet engraftment and prevent rejection in BM in the absence of immunosuppression. MethodsA model of BM local irradiation was set up. Islets were transplanted in syngeneic and fully major histocompatibility complex–mismatched recipients in control and locally irradiated BM; gain of normoglycemia and time to rejection were evaluated. ResultsBM local irradiation proved to be a selective and safe procedure. Syngeneic islet transplantation into locally irradiated BM had better outcome compared with not irradiated recipients in terms of capacity to gain normoglycemia (100% versus 56% in irradiated versus not irradiated mice). In the allogenic setting, glycemia was significantly lower in the first days after transplantation in the group of irradiated mice and local irradiation also delayed time to graft rejection (from 4 ± 1 days for not irradiated to 11 ± 1 days for locally irradiated mice). DiscussionThese data indicate that local immunosuppression by irradiation before islet transplantation in BM favors islet engraftment and delays time to rejection.

Effect of thalidomide on bone marrow angiogenesis in multiple myeloma patients

BackgroundBone marrow angiogenesis is increased in multiple myeloma (MM) patients, prompting the rationale for using antiangiogenic drugs in the treatment of these patients. ObjectiveTo assess angiogenesis in patients with MM at diagnosis and following treatment with an antiangiogenic drug. Patients and MethodsTwenty-three patients with newly diagnosed MM were treated with thalidomide-based regimens. Bone marrow evaluation was made before and following treatment and included angiogenesis assessment, which was quantified through microvessel density (MVD) determination, by means of anti-CD34 immunohistochemical labeling, and classified either as high MVD or low MVD, according to the mean CD34 count: above or below the median of 12.6. ResultsThe pre-therapy median MVD was 12 (7.5–18.3) versus 8.7 (5.35–18.5) post-therapy, p=0.2114. ConclusionsOur study found no reduction in MVD before and following treatment and, accordingly, we could establish no relationship between MVD and response to therapy in the sample we studied.

Screening differentially expressed proteins from co-cultured hematopoietic cells and bone marrow-derived stromal cells by quantitative proteomics (SILAC) method

BackgroundBone marrow stromal cells protect hematopoietic cells and provide drug resistance by delivering bunch of variable proteins. Thus, alterations of protein expression are typically associated with cell–cell signal transduction and regulation of cellular functions.MethodsCo-culture models of bone marrow stromal cells and hematopoietic cells are often used in studies of their crosstalk. Studies of altered protein expression initiated by stromal cell/hematopoietic cell interactions are an important new trend in microenvironmental research. There has been no report to date of global quantitative proteomics analysis of crosstalk between hematopoietic cells and stromal cells. In this study, we analyzed quantitative proteomes in a co-culture system of stromal HS5 cells and hematopoietic KG1a cells, and simultaneously tracked differentially expressed proteins in two types of cells before and after co-culture by stable isotope labeling by amino acids in cell culture (SILAC) method.ResultsWe have shown that in co-cultured KG1a, 40 proteins (including CKAP4, LMNA, and SERPINB2) were upregulated and 64 proteins (including CD44, CD99, and NCAM1) were downregulated relative to KG1a alone. We utilized IPA analysis to discover that the NOD-like receptor signaling pathway was upregulated, whereas platelet activation was downregulated in co-cultured KG1a cells. Furthermore, 95 proteins (including LCP1, ARHGAP4, and UNCX) were upregulated and 209 proteins (including CAPG, FLNC, and MAP4) were downregulated in co-cultured HS5 relative to HS5 alone. The tight junction pathway was downregulated and glycolysis/gluconeogenesis pathway was dysfunctional in co-cultured HS5. Most importantly, the significantly differentially expressed proteins can also be confirmed using different co-cultured cell lines.ConclusionAltogether, we recommend such quantitative proteomics approach for the studies of the hematopoietic–stroma cross-talk, differentially expressed proteins and related signaling pathways identification. The differentially expressed proteins identified from this current SILAC method will provide a useful basis for ongoing studies of crosstalk between stromal cells and hematopoietic cells in co-culture systems. All these result suggested our ongoing studies can focus on the mechanisms underlying CKAP4 increase and CD44 decrease in co-cultured hematopoietic cells, and the increase of LCP1 and decrease of CAPG in co-cultured stromal cell. The proteomic profiles from the KG1a/stromal cell co-culture system give new molecular insights into the roles of these cells in MDS pathophysiology and related bone disease.