Bone Marrow Research Articles

Current Concepts in Orthobiologics for Achilles Tendon Injuries: A Critical Analysis Review.

» Platelet-rich plasma and hyaluronic acid are low-risk and potentially high-reward treatments for Achilles tendinopathy, although clinical studies have yielded mixed results with questionable methodological quality» Case series and reports have reported that bone marrow aspirate, stem cells, and amniotic membrane products can improve functional outcomes, alleviate pain, and facilitate return to sport and activities, but high-level evidence studies are lacking» Exosomes are a promising novel biologic with laboratory studies showing improved collagen organization and cell proliferation, greater tendon mechanical properties, and prevention of extracellular matrix breakdown.» Standardization of protocols with clear reporting is necessary for future studies evaluating orthobiologic therapies for Achilles tendon injuries.

Neutrophil pyroptosis regulates corneal wound healing and post-injury neovascularisation.

The cornea is a unique structure that maintains its clarity by remaining avascular. Corneal injuries can lead to neovascularisation (CNV) and fibrosis and are the third most common cause of blindness worldwide. Corneal injuries induce an immune cell infiltration to initiate reparative processes. However, inflammation caused by sustained immune cell infiltration is known to be detrimental and can delay the healing process. This study was designed to understand the potential role of neutrophil and epithelial cell crosstalk in post-injury CNV. Western blotting and immunostaining assays demonstrated that neutrophils infiltrated corneas and underwent pyroptosis following acute alkali injury. In vivo studies showed that genetic ablation of Gasdermin D (GsdmD), a key effector of pyroptosis, enhanced corneal re-epithelialisation and suppressed post-injury CNV. In vitro co-culture experiments revealed that interleukin-1β (IL-1β) was released from pyroptotic neutrophils which suppressed migration of murine corneal epithelial cells. Real-time RT-PCR and immunostaining assays identified two factors, Wnt5a and soluble fms-like tyrosine kinase-1 (sflt-1), highly expressed in newly healed epithelial cells. sflt-1 is known to promote corneal avascularity. Bone marrow transplantation, antibody mediated neutrophil depletion, and pharmacological inhibition of pyroptosis promoted corneal wound healing and inhibited CNV in an in vivo murine corneal injury model. Taken together, our study reveals the importance of neutrophil/epithelium crosstalk and neutrophil pyroptosis in response to corneal injuries. Inhibition of neutrophil pyroptosis may serve as a potential treatment to promote corneal healing without CNV. Neutrophil pyroptosis delays re-epithelialization after corneal injury Compromised re-epithelialization promotes corneal neovascularization after injury Inhibition of post-injury pyroptosis could be an effective therapy to promote corneal wound healing.

Safety and low incidence of anaphylaxis in performing oral drug and food challenges in mastocytosis

BackgroundPatients with mastocytosis (MC) have an increased risk of severe anaphylaxis. They report hypersensitivity reactions to drugs and food, but causality often remains questionable. Most allergy centers avoid oral challenge tests (OCT) in MC patients. ObjectiveTo determine the safety of food and drug OCT in patients with MC. MethodsTo retrospectively evaluate the safety and outcome of challenges to drugs and food including food additives, out of 126 adult MC inpatients, 83 patients had a suspicion of food or drug hypersensitivity and 445 OCTs were performed. History, clinical data and allergy test results highlighting OCTs were analyzed. ResultsOnly 10 of 445 OCTs (2.2%) in 9 patients resulted in anaphylaxis. Drugs elicited reactions in 4/170 (2.4 %) patients: two patients to acetylsalicylic acid (2/39 tested patients, 5.1%), one to tramadol (1/12, 8.3%) and one to flurbiprofen (1/1). Anaphylaxis to food was recorded in 6/275 OCTs (2.2%); two out of 48 (4.1%) to α-gal, two to other foods and two to sulfites. Flushing or diarrhea occurred to histamine in 5/48 (10.4%), but also in 5/50 (10.0%) placebo challenges strongly questioning its relevance. Patients with proven anaphylaxis had more bone marrow MC and higher basal serum tryptase (71.3 vs. 44.3ug/l; p<0.05). ConclusionsChallenge-confirmed food and drug anaphylaxis was rare in MC patients. Results have to be interpreted cautiously as placebo reactions did occur. Severe anaphylaxis was seldom, but may occur and should be met by emergency preparedness.

Pansitopenia: Challenges Of Approach And Case Handling In Peripheral Hospitals

Pancytopenia is a hematological condition characterized by a decrease in three blood cell lines: erythrocytes, leukocytes, and platelets. This condition is not an independent disease, but a manifestation of various underlying diseases. Pancytopenia can be caused by impaired blood cell production in the bone marrow or increased destruction of blood cells in the periphery. Precise and prompt identification of the etiology is essential, given that pancytopenia can be life-threatening if not treated properly. This study aims to analyze and identify the causes of pancytopenia in patients at Batang Regency Hospital, as well as provide recommendations for appropriate treatment based on the etiology found. The methods used were clinical observation, laboratory examination, and diagnostic imaging in a 58-year-old patient with the main complaint of weakness and a history of pancytopenia. The examination includes a complete blood count, a smear of peripheral blood, and a clinical evaluation. The results showed that the patient had central type pancytopenia, with a suspected etiology of aplastic anemia or myelodysplastic syndrome. Other differential diagnoses include vitamin B12 deficiency. Although a bone marrow biopsy is necessary to confirm the diagnosis, initial therapy with blood transfusions and nutritional improvements has shown an improvement in the patient's condition. The conclusion of this study is the importance of a thorough evaluation to identify the etiology of pancytopenia. Proper treatment should be based on the underlying cause, which will affect the prognosis and prevent recurrence. A comprehensive approach can improve patients' quality of life and reduce the risk of complications

Identification of CD141+vasculogenic precursor cells from human bone marrow and their endothelial engagement in the arteriogenesis by co-transplantation with mesenchymal stem cells

BackgroundCritical limb ischemia (CLI) is a condition characterized by insufficient blood flow to the lower limbs, resulting in severe ischemia and potentially leading to amputation. This study aims to identify novel vasculogenic precursor cells (VPCs) in human bone marrow and evaluate their efficacy in combination with bone marrow-derived mesenchymal stem cells (BM-MSCs) for the treatment of CLI.MethodsEx vivo cultured VPCs and BM-MSCs from bone marrow were characterized and their effects on neovascularization and long-term tissue regeneration were tested in a mouse CLI model.ResultsVPCs, expressing high levels of hepatocyte growth factor and c-MET, were identified from human bone marrow aspirates. These cells exhibited strong vasculogenic capacity in vitro but possessed a cellular phenotype distinct from those of previously reported endothelial precursor cells in circulation or cord blood. They also expressed most surface markers of BM-MSCs and demonstrated multipotent differentiation ability. Screening of 376 surface markers revealed that VPCs uniquely display CD141 (thrombomodulin). CD141+VPCs are present in BM aspirates as a rare population and can be expanded ex vivo with a population doubling time of approximately 20 h, generating an elaborate vascular network even under angiogenic factor-deficient conditions and recruiting BM-MSCs to the network as pericyte-like cells. Intramuscular transplantation of a combination of human CD141+VPCs and BM-MSCs at a ratio of 2:1 resulted in limb salvage, blood flow recovery, and regeneration of large vessels in the femoral artery-removed CLI model, with an efficacy superior to that of singular transplantation. Importantly, large arteries and arterioles in dual cell transplantation expressed human CD31 in the intima and human α-smooth muscle actin in media layer at 4 and 12 weeks, likely indicating their lineage commitment to endothelial cells and vascular smooth muscle, respectively, in vivo.ConclusionDual-cell therapy using BM-derived CD141+ VPCs and BM-MSCs holds potential for further development in clinical trials to treat peripheral artery disease and diabetic ulcers.

The Prevalence of Blastocystis Hominis and Gastrointestinal System Parasites in Patients With Newly Diagnosed Multiple Myeloma Diagnosis

Introduction: Multiple myeloma (MM) is a hematological malignancy characterized by the presence of abnormal clonal plasma cells in the bone marrow. Immunodeficiency seen in patients with multiple myeloma (MM) is a serious problem. Blastocystis sp. is found in the gastrointestinal tract and can infect humans. Background/ Aims: In our study, we compared the prevalence of B.hominis and other parasites with the control group and patients who had recently been diagnosed with multiple myeloma. Material-Methods: Ninety-five multiple myeloma patients from our center and 95 volunteers as a control group were included in our study. The patients did not have any symptoms when stool samples were taken. For B.hominis, three consecutive stool samples were examined by saline-lugol trichrome staining and formalin-ethyl acetate concentration. Data were recorded and analyzed using SPSS 25.0. Results: Patients with newly diagnosed multiple myeloma and any of the patients in the control group did not have any parasites other than B.hominis. A significant difference was found between the newly diagnosed MM patients and the control group in terms of the number of B.hominis occurrences in the stool (p &amp;lt;0.001). Conclusions: According to this result, parasite load in stool increased in patients with MM. This result may reflect the suppression of the immune system of patients with MM. Accordingly, B.hominis should be considered as a causative agent in the presence of gastrointestinal symptoms that may occur during treatment in patients with MM.

Immunophenotyping myelodysplastic neoplasms: the role of flow cytometry in the molecular classification era.

The unique heterogenous landscape of myelodysplastic syndromes/neoplasms (MDS) has resulted in continuous redefinition of disease sub-entities, in view of the novel translational research data that have clarified several areas of the pathogenesis and the progression of the disease. The new international classifications (WHO 2022, ICC 2022) have incorporated genomic data defining phenotypical alterations, that guide clinical management of specific patient subgroups. On the other hand, for over a decade, multiparameter flow cytometry (MFC) has proven its value as a complementary diagnostic tool for these diseases and although it has never been established as a mandatory test for the baseline evaluation of MDS patients in international guidelines, it is almost universally adopted in everyday clinical practice for the assessment of suspected cytopenias through simplified scoring systems or elaborate analytical strategies for the detection of immunophenotypical dysplastic features in every hematopoietic cell lineage in the bone marrow (BM). In this review, we explore the clinically meaningful interplay of MFC data and genetic profiles of MDS patients, to reveal the currently existing and the potential future role of each methodology for routine clinical practice, and the benefit of the patients. We reviewed the existing knowledge and recent advances in the field and discuss how an integrated approach could lead to patient re-stratification and guide personalized management.

Epstein–Barr Virus encephalitis associated hemophagocytic lymphohistiocytosis in childhood-onset systemic lupus erythematosus: a case-based review

BackgroundHemophagocytic lymphohistiocytosis (HLH) is characterized by immune dysregulation that results in an uncontrolled hyperinflammatory state. HLH is classified into two main categories: primary (familial) HLH and secondary (acquired) HLH. Secondary HLH can result from various underlying, including infection-associated hemophagocytic syndrome (IAHS) and macrophage activation syndrome (MAS) associated with rheumatologic disorders, among others. Epstein–Barr virus (EBV) often causes IAHS, but central nervous system (CNS) involvement is rare among systemic lupus erythematosus (SLE) patients. We report a case of EBV encephalitis associated with HLH in a patient with childhood-onset SLE.Case presentationA 12-year-old girl had received a diagnosis of SLE 2 months before presentation. After a period of inactive disease on treatment, fever and seizures, with altered mental status and hallucinations, developed over several weeks. A complete blood cell count (CBC) revealed pancytopenia, accompanied by elevated levels of inflammatory markers: 86 mm/hr erythrocyte sedimentation rate, 8.9 mg/dl c-reactive protein, and 3,966 ng/mL of ferritin. The differential diagnosis included active neuropsychiatric SLE, CNS infection and neurological manifestations in secondary HLH, which could have represented either IAHS or MAS. Meropenem and acyclovir were initially administered for clinical acute encephalitis, followed by pulse methylprednisolone; however, the fever persisted, and another CBC revealed progressive cytopenia. A bone marrow study showed hypocellularity and active hemophagocytic activity, and intravenous immunoglobulin was additionally given due to the diagnosis of HLH. Cerebrospinal fluid (CSF) analysis showed 60/mm3 white blood cells (N 55%, L 45%), 141 mg/dL glucose (0.7 blood-CSF glucose ratio), < 4 mg/dL protein; results of Gram stain and bacterial culture were negative. The viral encephalitis panel from the CSF confirmed EBV infection. Bone marrow immunohistochemistry examination revealed increasing levels of CD8 + T-cell and equivocal positive results for EBV-encoded RNA in situ hybridization; therefore, HLH potentially associated with EBV was diagnosed. After treatment with IVIg, cyclosporin A, and prednisolone, the patient’s symptoms gradually improved and she was eventually able to return to school.ConclusionsOur case highlights the importance of a thorough differential diagnosis, including EBV encephalitis associated with HLH, in patients with childhood SLE, particularly in cases of clinical deterioration occurs after initial treatment.

Breast cancers that disseminate to bone marrow acquire aggressive phenotypes through CX43-related tumor-stroma tunnels.

Estrogen receptor-positive (ER+) breast cancer commonly disseminates to bone marrow, where interactions with mesenchymal stromal cells (MSCs) shape disease trajectory. We modeled these interactions with tumor-MSC co-cultures and used an integrated transcriptome-proteome-network-analyses workflow to identify a comprehensive catalog of contact-induced changes. Conditioned media from MSCs failed to recapitulate genes and proteins, some borrowed and others tumor-intrinsic, induced in cancer cells by direct contact. Protein-protein interaction networks revealed the rich connectome between 'borrowed' and 'intrinsic' components. Bioinformatics prioritized one of the 'borrowed' components, CCDC88A/GIV, a multi-modular metastasis-related protein that has recently been implicated in driving a hallmark of cancer, growth signaling autonomy. MSCs transferred GIV protein to ER+ breast cancer cells (that lack GIV) through tunnelling nanotubes via connexin (Cx)43-facilitated intercellular transport. Reinstating GIV alone in GIV-negative breast cancer cells reproduced approximately 20% of both the 'borrowed' and the 'intrinsic' gene induction patterns from contact co-cultures; conferred resistance to anti-estrogen drugs; and enhanced tumor dissemination. Findings provide a multiomic insight into MSC→tumor cell intercellular transport and validate how transport of one such candidate, GIV, from the haves (MSCs) to have-nots (ER+ breast cancer) orchestrates aggressive disease states.

In Vitro Purging of Acute Lymphoblastic Leukemia (B-ALL) Cells with the Use of PTL, DMAPT, or PU-H71.

Acute lymphoblastic leukemia (ALL) is a hematopoietic disorder that mainly affects the child population, and it is characterized by the presence of lymphoid progenitor or precursor cells with different genetic alterations. The origin of this disease is controversial, since some authors assumed that leukemic transformation occurs in a lymphoid progenitor, and there is also evidence that suggests the existence of leukemic initiating cells (LIC). PTL, DMAPT, and PU-H71 are agents that have been shown to eliminate bulk and stem cells from myeloid leukemias, but this effect has not been analyzed in lymphoblastic leukemias. In this study, we evaluated the effect of these compounds in different populations from pediatric B-ALL. For this, bone marrow samples from pediatric patients without treatment were obtained and cultured in the presence or absence of PTL, DMAPT, and PU-H71. The viability and apoptosis index were analyzed by flow cytometry in different hematopoietic subpopulations. These observations indicate that PTL and DMAPT are able to reduce B-ALL cells with a minimum effect in normal hematopoietic and non-hematopoietic cells. In contrast, PU-H71 was able to reduce the leukemic population and had a minimal effect in normal cells. These results present evidence that PTL and DMAPT are able to abrogate in vitro different populations of B-ALL and could represent a possibility of treatment, as well as prevent disease progression or relapse.

Biologic Augmentation of Rotator Cuff Repair: Current Concepts Review.

Rotator cuff tears are common in an aging population. Thus far, primary repairs have shown high re-tear rates suggesting the need for improved healing modalities. Current augmentations of rotator cuff repairs include synthetic and biological scaffolds, surgical bone marrow venting, and infusing the repair with a variety of stem cells and growth factors aimed at restoring the native cellular structure and function of the repaired tissue. This current concepts review discusses the anatomy, physical presentation, diagnosis, and treatment of rotator cuff tears; biological adjuvants for rotator cuff repairs; and the current literature on outcomes after biologically augmented rotator cuff repairs. [Orthopedics. 202x;4x(x):xx-xx.].

Construction and Evaluation of Hepatic Targeted Drug Delivery System with Hydroxycamptothecin in Stem Cell-Derived Exosomes.

Hydroxycamptothecin (HCPT) is commonly used in the treatment of liver cancer; however, its low water solubility and poor stability significantly limit its clinical application. In recent years, research on exosomes has deepened considerably. Exosomes possess a unique phospholipid bilayer structure, enabling them to traverse tissue barriers, which provides natural advantages as drug carriers. Nevertheless, delivering exosomes safely and efficiently to target cells remains a major challenge. In this study, we utilized the affinity of the SP94 peptide for human liver cancer cell receptors. HCPT was coated with exosomes in our experimental design, and the exosome membrane was modified with SP94 peptide to facilitate drug delivery to liver cancer cells. Exosomes were purified from bone marrow mesenchymal stem cells, and targeted peptides were attached to their surfaces via post-insertion techniques. Subsequently, HCPT was incorporated into the exosomes through electroporation. Using the HepG2 hepatoma cell line, we evaluated a series of in vitro pharmacodynamics and studied pharmacokinetics and tissue distribution in animal models. The results indicated that ligand-targeted, modified drug-carrying exosomes significantly enhance drug bioavailability, prolong retention time in vivo, and facilitate liver targeting. Moreover, this approach reduces drug nephrotoxicity, enhances anti-tumor efficacy, and lays the groundwork for the development of novel liver cancer-targeting agents.

Bilateral exudative retinal detachment with subretinal light-chain protein in a patient with multiple myeloma -case report-

BackgroundExudative retinal detachment in patients with multiple myeloma is exceedingly rare. Only two Cases are known to us. We successfully identified light-chain proteins in subretinal fluid, allowing for a more precise understanding of the pathogenesis of this complication.Case presentationA 68-year-old patient presented with bilateral exudative retinal detachment. The visual impairment was reported one day after stem cell mobilization by granulocyte-colony stimulating factor (G-CSF) and the additional administration of Plerixafor. The symptoms began during stem cell apheresis. The patient underwent surgical treatment for both eyes through vitrectomy and silicone oil tamponade. Light-chain proteins were detected in the collected subretinal fluid through electrophoresis in one eye.ConclusionsWe successfully identified light-chain proteins in subretinal fluid, allowing for a more precise understanding of the pathogenesis of this complication. The pathomechanism likely involves damage to the outer blood-retina barrier due to the deposition of light-chain proteins. Whether mobilization of bone marrow cells with Plerixafor led to a breakdown of the outer blood-retina barrier in these patients is a topic for discussion and has to be considered in the use of Plerixafor.

From Cord to Cure: Repurposing Biological Waste for UAE's Health Revolution with Umbilical Cord Stem Cell Therapy

In the UAE, current hospital policies mandate the disposal of biological waste, such as umbilical cords. However, this tissue represents a valuable source of therapeutic material like mesenchymal stem cells (MSCs) and their secretome. The disposal of this valuable therapeutic material imposes a significant burden on public health. In the United States alone, approximately 18,000 people per year are diagnosed with diseases that could be ameliorated with stem cell therapy. These individuals often remain untreated due to the lack of available stem cells, as current methods for MSC obtention, such as Bone Marrow-MSCs, require an invasive bone marrow aspirate procedure, which is painful and can pose risks to donors, including infection and prolonged recovery times. This paper advocates for policy reform to facilitate the collection, storage, and utilization of umbilical cords. By addressing regulatory barriers and implementing standardized protocols, hospitals in the UAE can not only contribute to advancements in regenerative medicine but also solidify the nation’s position as a leader in this emerging field, ultimately improving patient outcomes and reducing the disease burden both locally and regionally. This is particularly relevant given the significant burden of cardiovascular diseases and type 2 diabetes mellitus (T2DM) in the UAE, where 17.3% of the population aged 20 to 79 is affected with T2DM. Cardiovascular diseases and T2DM are few of the many disorders that can benefit from MSC therapy. Utilizing umbilical cord biowaste for such diseases can further build the UAE’s status as a hub for medical tourism and accentuate it’s growing prominence in the field of regenerative medicine.

TARDBP drives T-cell acute lymphoblastic leukemia progression by binding MDM2 mRNA, involving β-catenin pathway.

T-cell acute lymphoblastic leukemia (T-ALL) is a dangerous hematological malignancy. The trans-activation response DNA binding protein (TARDBP), an RNA/DNA binding protein, is involved in the growth and metastasis of multiple cancers. However, TARDBP has not been reported in T-ALL. It was found that TARDBP was highly expressed in pediatric T-ALL samples by microarray GSE26713 (log2 fold change >1, p < .05). Herein, TARDBP was silenced and overexpressed by lentivirus transduction in T-ALL cell lines, including Jurkat and Molt4 cells. Invitro, silencing TARDBP inhibited T-ALL cell proliferation and cycle progression and accelerated cell apoptosis, while overexpressing TARDBP induced the opposite effects. In addition, we investigated whether the β-catenin pathway could be activated by TARDBP in T-ALL cells. Moreover, XAV-939, a β-catenin inhibitor, was capable of suppressing the malignant phenotypes in TARDBP-overexpressed T-ALL cells. Invivo, TARDBP-silenced or TARDBP-overexpressed T-ALL cells were injected into mice. We found that TARDBP promoted T-ALL cell growth in the spleens and bone marrows of mice. On the basis of GSE26713, there was a significant correlation between TARDBP and mouse double minute 2 (MDM2). The RIP-PCR assay demonstrated that TARDBP bound MDM2 mRNA in T-ALL cells. The rescue experiments further revealed the roles of the TARDBP/MDM2 axis in T-ALL cell phenotypes, which was also reflected by mRNA-seq. In aggregate, we explored a promising biomarker, TARDBP, for T-ALL treatment. The underlying mechanisms might involve the interaction with MDM2 mRNA and the regulation of the β-catenin pathway.

Whole Blood Transcriptome Analysis in Congenital Anemia Patients.

Congenital anemias include a broad range of disorders marked by inherent abnormalities in red blood cells. These abnormalities include enzymatic, membrane, and congenital defects in erythropoiesis, as well as hemoglobinopathies such as sickle cell disease and thalassemia. These conditions range in presentation from asymptomatic cases to those requiring frequent blood transfusions, exhibiting phenotypic heterogeneity and different degrees of severity. Despite understanding their different etiologies, all of them have a common pathophysiological origin with congenital defects of erythropoiesis. We can find different types, from congenital sideroblastic anemia (CSA), which is a bone marrow failure anemia, to hemoglobinopathies as sickle cell disease and thalassemia, with a higher prevalence and clinical impact. Recent efforts have focused on understanding erythropoiesis dysfunction in these anemias but, so far, deep gene sequencing analysis comparing all of them has not been performed. Our study used Quant 3' mRNA-Sequencing to compare transcriptomic profiles of four sickle cell disease patients, ten thalassemia patients, and one rare case of SLC25A38 CSA. Our results showed clear differentiated gene map expressions in all of them with respect to healthy controls. Our study reveals that genes related to metabolic processes, membrane genes, and erythropoiesis are upregulated with respect to healthy controls in all pathologies studied except in the SLC25A38 CSA patient, who shows a unique gene expression pattern compared to the rest of the congenital anemias studied. Our analysis is the first that compares gene expression patterns across different congenital anemias to provide a broad spectrum of genes that could have clinical relevance in these pathologies.

Is Bone Marrow Metastasis in Gastric Cancer Adequate for Best Supportive Care Decisions? Or is There Still a Chance?

Is Bone Marrow Metastasis in Gastric Cancer Adequate for Best Supportive Care Decisions? Or is There Still a Chance?

Increased neutrophil extracellular traps caused by diet-induced obesity delay fracture healing.

Obesity, recognized as a risk factor for nonunion, detrimentally impacts bone health, with significant physical and economic repercussions for affected individuals. Nevertheless, the precise pathomechanisms by which obesity impairs fracture healing remain insufficiently understood. Multiple studies have identified neutrophil granulocytes as key players in the systemic immune response, being the predominant immune cells in early fracture hematomas. This study identified a previously unreported critical period for neutrophil infiltration into the callus. Invivo experiments demonstrated that diet-induced obesity (DIO) mice showed earlier neutrophil infiltration, along with increased formation of neutrophil extracellular traps (NETs), compared to control mice during the endochondral phase of fracture repair. Furthermore, Padi4 knockout was found to reduce NET formation and mitigate the fracture healing delays caused by high-fat diets. Mechanistically, invitro analyses revealed that NETs, by activating NLRP3 inflammasomes, inhibited the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and concurrently promoted M1-like macrophage polarization. These findings establish a connection between NET formation during the endochondral phase and delayed fracture healing, suggesting that targeting NETs could serve as a promising therapeutic approach for addressing obesity-induced delays in fracture recovery.

Triple immunostaining demonstrates the possible existence of segregated-nucleus-containing atypical monocytes in human primary myelofibrosis bone marrow: a case report

BackgroundSegregated-nucleus-containing atypical monocytes have recently been identified in mice. Segregated-nucleus-containing atypical monocytes are thought to originate from the bone marrow and induce fibrosis in the drug-injured lung. The Lyc6c− murine monocyte subset is the counterpart to human CD14−CD16++ non-classical monocytes; however, the human counterpart to murine segregated-nucleus-containing atypical monocytes has not yet been identified. Primary myelofibrosis is a well-known disease of progressive marrow fibrosis, and atypical megakaryocytes are thought to be closely related to fibrosis in primary myelofibrosis bone marrow. However, recently, monocytes have been reported to play an important role in marrow fibrosis in primary myelofibrosis. We speculated that, if there is a human counterpart to murine segregated-nucleus-containing atypical monocytes, it would present the same markers as murine segregated-nucleus-containing atypical monocytes, such as CD14−CD16+ macrophage-1 antigen (CD11b/CD18 complex)+, MSR1+, and CEACAM1+, and it might exist in the bone marrow of patients with primary myelofibrosis.Case presentationA 74-year-old Japanese male visited our hospital for clinical follow-up after total prostatectomy for prostatic cancer. Anemia, thrombocytosis, and elevated lactate dehydrogenase were suddenly observed in a periodic examination. CALR mutation type 2 (p.K385fs*47) was observed. The histological features of the patient’s bone marrow were consistent with fibrotic primary myelofibrosis. We immunohistochemically studied the bone marrow in an attempt to identify a human counterpart to murine segregated-nucleus-containing atypical monocytes. We detected a few CD16+MSR1+CEACAM1+ cells, but not CD14+MSR1+CEACAM1+ cells, by triple immunostaining. The patient is in a good condition and does not require treatment for primary myelofibrosis.ConclusionThere is a possibility that human segregated-nucleus-containing atypical monocytes exist in the bone marrow of primary myelofibrosis patients and might be related to marrow fibrosis.

The stem and progenitor cells and the functional activity of liver from age-different Wistar rats

The liver has a big potential for self-healing, but the activity of regeneration decreases with age. Changes are occurring, including in the functional activity of various liver cell populations, the study of the characteristics of which can become the basis for the development of new therapeutic approaches to the liver diseases treatment at older people. The aim of this research was to study the level of stem and progenitor cells and the functional activity of the healthy liver from age- different rats. Material and methods. Experiments were carried out on Wistar rats aged 6 and 12 months. Ultrasound and histological examination of the liver from rats was used to assess morphological changes. The lipid profile of blood serum was evaluated by biochemical methods. Cytometric methods were used to study the surface and intracellular antigens of stem and progenitor cells isolated from the bone marrow, arterial blood and liver of rats. Results and discussion. In 12-month-old male Wistar rats, compared with 6-month-old rats, excessive formation of extracellular matrix components, disruption of tissue architecture, development of portal hypertension, as well as an increase in the concentration of cholesterol, triglycerides, high- and low-density lipoproteins were revealed. We identified age- related differences in the content of hematopoietic and mesenchymal stem cells, epithelial cells (CD45–CD326+) in the bone marrow, blood and liver of rats. In the liver parenchyma, the populations of hepatocyte precursors (CD45– CD326+CD133+), oval cells (CD45–CD326+CD133+CD90+). At the same time, the level of all cell populations in the liver parenchyma of rats expressing the intracellular marker Sox9 was higher in one-year-old animals compared to younger ones, regardless of the cell phenotype. Conclusions. In the liver of 12-month-old rats, compared to 6-month-old rats, the number of cells expressing Sox9, lymphocytes with an inflammatory phenotype increases, the number of stem cells and various populations of epithelial and endothelial cells decreases, which leads to a decrease in the regenerative capacity of the liver, disruption of the tissue architecture of the organ and changes in lipid metabolism. These changes largely determine the increased susceptibility with age to the development of chronic liver diseases.