Marrow Proliferation Research Articles

Clinical and immunological characteristics of high-risk double-hit multiple myeloma

At present, the characteristics of double-hit multiple myeloma (DHMM) are unknown. We retrospectively analyzed the clinical data from 433 new diagnosed MM patients and found that DHMM have a higher β2-MG level and percentage of bone marrow plasma cell. Cox regression analysis showed that the prognosis of DHMM was not limited by clinical indicators. The abnormal proliferation of bone marrow in DHMM is obvious, and the proportion of poorly differentiated plasma cell is high. By collecting specimens from our center and performing flow cytometry to analyze the immunophenotypic and functional characteristics of lymphocyte subpopulations, we found that DHMM had a higher ratio of Tregs cells, and the proportion of iTregs cells was also significantly higher than non-DHMM (P < 0.05). Moreover, DHMM had higher levels of TGF-β1 and IL-10, and TGF-β1 and IL-10 were positively correlated with iTregs (P < 0.05). In addition, DHMM was highly expressed PD-1 on CD8 + T cells and had a higher proportion of CD38highTregs cells. In vitro we have shown that the addition of TGF-β1 antibody or CD38 antibody can effectively inhibit the proportion of CD38high Tregs. This study describes the characteristics of DHMM based on bicentric data, which is helpful to better provide theoretical support for the treatment of DHMM.

Characteristics of DNMT3a mutation in acute myeloid leukemia and its prognostic implication

BackgroundAcute myeloid leukemia (AML) is a clonal disorder arising from the differentiation arrest of myeloid precursor and malignant proliferation of a bone marrow derived, self-renewing stem or progenitor cells inside the bone marrow (BM) and blood due to numerous genetic mutations. Some mutations can also adjust DNA methylation and may play a critical function in pathogenesis in Cytogenetically Normal Acute Myeloid Leukemia (CN-AML). Somatic mutations in DNMT3a were pronounced in approximately 20% and ∼30–35% of overall AML and CN-AML, respectively. Most DNMT3a mutations in AML have been observed to be heterozygous, A missense mutation, R882, located inside Hot spot exon 23, has been found to be the maximum common mutation. This is a preliminary study conducted on 20 adult Egyptian patients newly diagnosed as AML where Sanger sequencing of Hotspot Exon 23 of DNMT3a gene was performed on their initial bone marrow samples and were followed up to 3 months post-induction therapy. Only De Novo AML patients were included in our study.ResultsOur results revealed that overall DNMT3a mutations were present in 25% of our patients, 10% having the R882 (rs147001633) mutation being 5% R882C and 5% R882H. Immunophenotyping analysis among Mutated DNMT3a (R882 and Non R882) and Wild DNMT3a revealed that AML markers exhibited no significant differences except for myeloperoxidase positivity which was significant among the groups (0.050). Regarding cytogenetics, only one case of the mutated DNMT3a had positive FISH inv (16), where the rest were FISH negative. After 28 days of induction, 75% of all our patients achieved complete response (CR), 20% achieve partial response (PR) out of which 75% are DNMT3a mutated. After 3 months follow-up, 10% of all patients faced mortality where 5% was DNMT3a wild type (died due to treatment-related mortality) and 5% was R882 mutated DNMT3a.ConclusionDNMT3a mutations are present in 25% (5/20) of our AML patients, with 10% (2/20) having the R882 mutation being 5% (1/20) R882C and 5% (1/20) R882H. R882 mutation is associated with resistance to chemotherapy, and poorer outcomes, highlighting its poorer prognostic significance in AML.

Dose-Response Relationships Between Radiation Exposure, Bone Marrow Function as Measured by 18F-Fluorothymidine Positron Emission Tomography, and Lymphocyte Counts During Chemoradiation for Non-Small Cell Lung Cancer

Purpose18F-Fluorothymidine(FLT)-PET enables sensitive imaging of bone marrow (BM) proliferation. Sequential FLT-PET/CT scans before and during chemoradiation (CRT) for non-small cell lung cancer (NSCLC) were repurposed to investigate dose-response effects of radiation on BM proliferation. Methods and materialsTwenty-six NSCLC patients underwent platinum-based CRT to 60Gy in 30 fractions with FLT-PET/CT scans at baseline, week 2 (20Gy) and week 4 (40Gy). FLT uptake in BM was isolated using Medical Image Merge software. Week 2 and week 4 FLT-PET BM scans were fused with contemporaneous radiation isodose distributions. Relationships between radiation dose and FLT BM uptake (SUVmax and visual parameters) were analyzed using generalized-linear and restricted cubic-spline models. Percentage volumes of total BM without appreciable FLT uptake (“ablated”) on week 2 and week 4 FLT-PET scans were calculated by comparisons with baseline scans. ResultsThoracic FLT uptake was ablated in BM regions exposed to cumulative radiation doses ≥3Gy by week 2. In all cases BM FLT SUVmax declined rapidly as radiation dose increased. BM proliferation significantly decreased by more than 95% after ≥3-4Gy at 2 weeks and ≥4-5Gy at 4 weeks. The ablated BM volume increased from week 2 to week 4 as BM in the penumbra accumulated radiation dose. Median percentage of total BM ablated was 13.1% (range 5.6-20.3) at 2 weeks and 15.7% (range 9.2 - 24.1) at 4 weeks. Mean lymphocyte counts fell from baseline of 2.01×109/L to 0.77 week 2, and 0.60 week 4. Lymphocyte decline strongly correlated with percentage of total BM ablated by week 4 (y=-46 -1.64 x, R2adj = 0.34, p=0.001). ConclusionsBM ablation associated with low dose-radiation exposure during CRT correlated significantly with lower week 4 lymphocyte counts. BM is a potential organ-at-risk and reducing the BM volume exposed to ≥3Gy may help preserve lymphocytes essential for effective adjuvant immunotherapy.

Tartrate-resistant acid phosphatase (TRAP/ACP5) promotes bone length, regulates cortical and trabecular bone mass, and maintains growth plate architecture and width in a sex- and site-specific manner in mice

Tartrate-resistant acid phosphatase (TRAP) serum levels reflect osteoclast number, bone remodeling activity, and fracture risk. Deletion or loss of function of TRAP results in short stature in mice and man. Yet, the impact and mechanisms of TRAP for the site- and sex-specific development of bone and cartilage is not well understood. Here, we use a global TRAP knockout (TRAPKO) and wildtype littermate control (WT) mice of both sexes to investigate TRAP as a possible sex- and site-specific regulator of bone and growth plate development. TRAPKO mice of both sexes weighed less and had shorter tibial length than their WT, features that were more accentuated in male than female TRAPKO mice. These changes were not associated with a general reduction in growth as not all organs displayed a proportionally lower mass, and serum IGF-1 was unchanged. Using μCT and site-specificity analysis of the cortical bone revealed wider proximal tibia, a higher trabecular thickness, and lower trabecular separation in male TRAPKO compared to WT mice, an effect not seen in female mice. Histomorphometric analysis revealed that the growth plate height as well as height of terminal hypertrophic chondrocytes were markedly increased, and the number of columns was decreased in TRAPKO mice of both sexes. These effects were more accentuated in female mice. Proliferation and differentiation of bone marrow derived macrophages into osteoclasts, as well as C-terminal cross links were normal in TRAPKO mice of both sexes. Collectively, our results show that TRAP regulates bone and cartilage development in a sex-and site-specific manner in mice.

Electrochemical Mineralization Regulates Hydroxyapatite Deposition of Silk Fibroin Nanofibers for Promoting Osteogenic Differentiation of Human Mesenchymal Stem Cells

ABSTRACTScaffold and stem cells are the key elements in the procedure of bone repair. Bombyx mori silk fibroin (SF) is the proper material for bone tissue engineering due to its biocompatibility and its easy to obtain nanofiber structure. The suitable cell substrate is also an important factor because different cells have different adaptations to composite. For this case, selecting a scaffold that can promote osteogenic differentiation of various cells is crucial and meaningful. In this work, hydroxyapatite (HA) was deposited precisely onto each silk nanofiber by electrochemical mineralization (EC) to form SF/HA. SF/HA can promote the proliferation and osteogenic differentiation of both human bone marrow mesenchymal‐derived stem cells (hMSCs) and human adipose‐derived mesenchymal stem cells (hAMSCs). It also promot the expression of alkaline phosphatase (ALP) and osteogenic differentiation related genes. Western blot analyses show mitogen‐activated protein kinase (MAPK) signal pathway is regulated by SF/HA. Therefore, the study provides a proper method to obtain a good composite SF/HA and it promotes the osteogenic differentiation of both hMSCs and hAMSCs.

Clinical Characteristics and Diagnosis of Nonaccelerating MDS/MPN-U Patient with Granulocyte Dysplasia.

The goal was to improve the clinical cognition of nonaccelerating myelodysplastic/myeloproliferative neoplasms-unclassifiable (MDS/MPN-U) and avoid misdiagnosis or delayed diagnosis. The clinical manifestations, laboratory indicators, histopathology, and therapeutic effects of a patient with nonaccelerating MDS/MPN-U were analyzed and the relevant literature was reviewed. Blood routine: white blood cell 98.48 x 109/L, red blood cell 3.20 x 1012/L, basophils 0.42 x 109/L, eosinophils 1.31 x 109/L, hemoglobin 112 g/L, and platelet 113 x 109/L. Blood smears showed granulocytosis and cells at various stages, polylobular granulocytes also can be seen. Bone marrow images show granulocytosis and dysplastic neutrophils, such as binuclear granulocyte, cyclic nuclear granulocyte, nuclear punch, cytoplasm vacuoles, polylobular granulocytes and so on. Bone marrow biopsy: Bone marrow proliferation tumor, combined with cell morphology and molecular biochemistry is recommended. Gene test showed Jak-2 positive, BCR/ABL and MPL negative. Chromosome examination indicated the presence of 46, XY, add (2)(p25), del (12) (p11.2p13)[16]/46, XY. MDS/MPN-U with granulocytosis and dysplastic neutrophils is rare, mostly in the elderly, and the diagnosis should be made except for other myeloid tumors. Currently, there is no uniform treatment guideline or expert consensus. The treatment options are limited and need to be further confirmed by more studies. MDS/ MPN-U with granulocytosis and dysplastic neutrophils has adverse prognostic factors such as advanced age, increase of bone marrow original cells and related gene mutations. Whether the adverse prognosis is related to specific gene mutations and cytogenetic variation remains to be clarified by more research data.

Polysaccharide hybrid scaffold encapsulated endogenous factors for microfracture enhancement by sustainable release and cell recruitment

Autologous matrix-induced chondrogenesis improves microfracture (MF), but the lack of functional cells and factors inhibit a curative effect. Here, inspired by the healing process of wounds, we prepared a polysaccharide hybrid scaffold (HDCP) to immobilize endogenous growth factors (eGFs), which were derived from platelet-rich plasma (PRP). Compared with the absence of PRP, the hybrid cross-linking network endowed this scaffold with a denser pore structure, enhanced mechanical properties, and slower protein mass transfer rate, thus ensuring the long-term sustainable release of eGFs. In vitro studies confirmed that HDCP promoted bone marrow mesenchymal stem cell recruitment, proliferation, and chondrogenic differentiation. In vivo implantation indicated that HDCP regenerated a smooth articular cartilage surface with a regular fibrous matrix arrangement structure, and the mechanical strength of the regenerated cartilage considerably improved. The obvious MF enhancement implied that PRP and polysaccharide scaffold hybridization could be used as an implantable matrix for cartilage repair.

In vitro evaluation of BMSCs early proliferation on minocycline-loaded electrospun nanofibers membrane.

Electrospun nanofibers could simulate the natural extracellular matrix (ECM) of the host bone, while minocycline (MINO) is a broad-spectrum tetracycline antibiotic which has been found to have multiple non-antibiotics biological effects that promotes osteogenesis in vitro and in vivo. The present study aims at constructing a polylactic acid (PLA) electrospun nanofiber membrane loaded with MINO to enhance Bone marrow mesenchymal stem cells (BMSCs) adhesion and proliferation for early clinical treatment. The MINO-PLA membrane were characterized by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR) and in vitro drug release study. The antibacterial ability was also investigated. In addition, in vitro cellular proliferation experiment was performed to verify whether the PLA electrospun nanofibers membrane loaded with MINO enhance BMSCs adhesion and proliferation. Analyzing the drug release and cell growth results, it was found that only the effective concentration of MINO-PLA could help the growth of BMSCs in the short term. This is related to the drug release rate of MINO-PLA and the initial concentration of MINO. This study shows that by controlling the concentration and release rate of MINO with electrospinning PLA, BMSCs could proliferate on it, and a new bone repair material had been made in this study.

Role of ITGA5 in Malignant Proliferation and Infiltration of Acute Myeloid Leukemia

Abstract I ntroduction : Acute myeloid leukemia is a clonal hematopoietic stem cell malignancy, and excessive leukemic cell migration into the peripheral blood can lead to fatal complications when leukemic cells in the bone marrow proliferate and infiltrate malignantly. Integrins are a class of adhesion molecules expressed on hematopoietic cells and leukemia cells. At present, it is believed that the interaction between integrins and their ligands also plays an important role in the migration and adhesion of leukemia cells. We performed proteomic sequencing analysis of AML bone marrow samples in our previous work and found that integrin ITGA5 expression was significantly increased in AML leukemia cells compared with normal subjects, and combined with previous work basis and literature reports we speculated that integrin molecule ITGA5 may be involved in the process of malignant proliferation and infiltration of AML. Methods: We collected bone marrow cells from AML patients and healthy controls for single-cell transcriptome and proteomics sequencing.Bone marrow CD34 + cells and AML cell lines from normal subjects and AML patients were collected, and integrin gene level expression was verified by PCR.We knocked down the ITGA5 gene in AML cell lines and analyzed ITGA5 expression by flow cytometry.AML cell lines were treated with anti-human ITGA5 monoclonal antibody and ITGA5 inhibitor ATN161, respectively, to determine the effects on apoptosis, migration, invasion, and adhesion, in addition, the surface molecular force of the cell lines was determined by a novel molecular fluorescence tension probe based on DNA structure.Using the MLL-AF9 mouse model, we tested the effect of blocking ITGA5 in combination with cytarabine chemotherapy.The doses of mice in each group were as follows: ATN161 (50 mg/kg) + cytarabine (50 mg/kg), ATN161 (50 mg/kg), cytarabine (50 mg/kg), and normal saline. Results: Proteomic sequencing analysis of bone marrow samples revealed that integrin ITGA5 expression was significantly increased in AML leukemia cells compared with normal subjects.RNA were extracted from AML cell lines Kasumi-1, MV411, MOLM-13, and bone marrow CD34 + cells from AML patients and detected by QPCR, and ITGA5 expression was significantly increased in AML cell lines and AML patient cells compared with normal controls.ITGA5 expression in AML cells was inhibited by either ITGA5 monoclonal antibody or ITGA5 inhibitor ATN161, and migration and invasion of AML cell lines were significantly decreased, but the effect on apoptosis was not very prominent.Using a novel molecular fluorescent tension probe based on DNA structure to visualize the mechanical signal of viable cells, AML cell lines, AML patients CD34 + can produce a strong signal on the base compared with controls, and the force signal generated on the base is significantly inhibited after the use of ATN161.We constructed a MLL-AF9 leukemia mouse model and found that ATN161 could significantly reduce the proportion of leukemic cells in the liver, spleen, and bone marrow of leukemia mice and effectively alleviate the malignant proliferation and infiltration of leukemia cells by ITGA5 inhibitor ATN161 combined with cytarabine treatment, and the therapeutic effect was more obvious after combination therapy. C onclusion : ITGA5 plays an important role in the migration, invasion and adhesion of leukemia cells. Inhibition of ITGA5 by ATN16 combined with chemotherapy can significantly play a better therapeutic role, and inhibit extramedullary infiltration of leukemia cells. Our findings will contribute to a better understanding of the role of ITGA5 in malignant proliferative infiltration of AML and the role of ITGA5 as a preclinical target in AML. Disclosures No relevant conflicts of interest to declare.

A novel degradable PCL/PLLA strapping band for internal fixation of fracture

Early fracture fixation is the critical factor in fracture healing. Common internal fracture implants are made of metallic materials, which often affects the imaging quality of CT and MRI. Most patients will choose secondary surgery to remove the internal fixation implants, which causes secondary damage to them. The development of new degradable internal fracture implants has attracted more and more attention from orthopedic surgeons and researchers. Based on these problems, we improved the various properties of medical grade polycaprolactone (PCL) by adding poly(L-lactide) (PLLA). We produced PCL/PLLA strapping bands with different mass ratios by injection molding. We compared the mechanical properties, degradation properties, cell biocompatibility, bone marrow mesenchymal stem cells (BMSCs) adhesion, proliferation, osteogenic differentiation and fracture fixation effect of these strapping bands. The results showed that the tensile strength and yield force of the strapping bands increased with the increase of the content of PLLA. The addition of PLLA could significantly improve the mechanical strength in the early stage and accelerate the degradation rate of the strapping band. PCL/PLLA (80/20) strapping band had no significant cytotoxicity toward rBMSCs and could promote osteogenic differentiation of rBMSCs. The strapping band could ensure femoral fracture healing of beagles in 3 months and didn’t cause damage to the surrounding tissues and main organs. This study will provide some new insights into the biodegradable products of PCL/PLLA blends for internal fixation of fracture.Graphical We produced novel degradable PCL/PLLA strapping bands with different mass ratios by injection molding. We tested the biological safety of the prepared internal fixation strapping bands for fracture, such as cell experiment in vitro and animal experiment, and studied the degradation behavior in vitro. The strapping bands could ensure femoral fracture healing of beagles. This study will provide some new insights into the biodegradable products of PCL/PLLA blends for internal fixation of fracture. A Immunofluorescence staining of rBMSCs (live cells: green; dead cells: red). B Young’s modulus change curve during strapping bands degradation. C The implantation process of strapping bands. D Micro-CT images of the beagle’s fracture recovery after the operation.

P1.28-05 Cessation of Marrow Proliferation on FLT-PET with Low RT doses during chemoRT for NSCLC Correlates with Lymphocyte Decline

P1.28-05 Cessation of Marrow Proliferation on FLT-PET with Low RT doses during chemoRT for NSCLC Correlates with Lymphocyte Decline

Expression Pattern and Prognostic Implication of SALL4 Gene in Acute Myeloid Leukemia

Abstract Background Acute myeloid leukemia (AML) is a clonal disorder arises from the differentiation arrest of myeloid precursor and malignant proliferation of a bone marrow derived, self-renewing stem or progenitor cells in the bone marrow (BM) and blood. Aim of the Work To characterize the expression pattern of SALL4 gene in patients with acute myeloid leukemia and to determine its prognostic impact. Patients and Methods This study was conducted on 20 Egyptian AML patients (recruited from hematology units of Internal Medicine department, Ain shams University Hospitals) and 10 patients as controls. All patients were subjected to full history taking, thorough clinical examination, complete blood count, BM aspiration cytochemical analysis, flow-cytometric immunophenotyping of BM blasts using acute leukemia panel and cytogenetic analysis using FISH technique and RT-PCR to measure SALL4 gene expression in AML cases and controls. Results Our results showed highly significant difference between expression of SALL 4 gene in AML patients than control group with RQ median in AML cases 15.46 and in control group 0.4 (p value 0.000). Non significant results about prognosios was reached Conclusion SALL4 gene expression is constitutive in De-novo AML cases.

Nacre-mimetic cerium-doped nano-hydroxyapatite/chitosan layered composite scaffolds regulate bone regeneration via OPG/RANKL signaling pathway

Autogenous bone grafting has long been considered the gold standard for treating critical bone defects. However, its use is plagued by numerous drawbacks, such as limited supply, donor site morbidity, and restricted use for giant-sized defects. For this reason, there is an increasing need for effective bone substitutes to treat these defects. Mollusk nacre is a natural structure with outstanding mechanical property due to its notable “brick-and-mortar” architecture. Inspired by the nacre architecture, our team designed and fabricated a nacre-mimetic cerium-doped layered nano-hydroxyapatite/chitosan layered composite scaffold (CeHA/CS). Hydroxyapatite can provide a certain strength to the material like a brick. And as a polymer material, chitosan can slow down the force when the material is impacted, like an adhesive. As seen in natural nacre, the combination of these inorganic and organic components results in remarkable tensile strength and fracture toughness. Cerium ions have been demonstrated exceptional anti-osteoclastogenesis capabilities. Our scaffold featured a distinct layered HA/CS composite structure with intervals ranging from 50 to 200 μm, which provided a conducive environment for human bone marrow mesenchymal stem cell (hBMSC) adhesion and proliferation, allowing for in situ growth of newly formed bone tissue. In vitro, Western-blot and qPCR analyses showed that the CeHA/CS layered composite scaffolds significantly promoted the osteogenic process by upregulating the expressions of osteogenic-related genes such as RUNX2, OCN, and COL1, while inhibiting osteoclast differentiation, as indicated by reduced TRAP-positive osteoclasts and decreased bone resorption. In vivo, calvarial defects in rats demonstrated that the layered CeHA/CS scaffolds significantly accelerated bone regeneration at the defect site, and immunofluorescence indicated a lowered RANKL/OPG ratio. Overall, our results demonstrate that CeHA/CS scaffolds offer a promising platform for bone regeneration in critical defect management, as they promote osteogenesis and inhibit osteoclast activation.

3D Printing of Microenvironment-Specific Bioinspired and Exosome-Reinforced Hydrogel Scaffolds for Efficient Cartilage and Subchondral Bone Regeneration.

In clinical practice, repairing osteochondral defects presents a challenge due to the varying biological properties of articular cartilages and subchondral bones. Thus, elucidating how spatial microenvironment-specific biomimetic scaffolds can be used to simultaneously regenerate osteochondral tissue is an important research topic. Herein, a novel bioinspired double-network hydrogel scaffold produced via 3D printing with tissue-specific decellularized extracellular matrix (dECM) and human adipose mesenchymal stem cell (MSC)-derived exosomes is described. The bionic hydrogel scaffolds promote rat bone marrow MSC attachment, spread, migration, proliferation, and chondrogenic and osteogenic differentiation in vitro, as determined based on the sustained release of bioactive exosomes. Furthermore, the 3D-printed microenvironment-specific heterogeneous bilayer scaffolds efficiently accelerate the simultaneous regeneration of cartilage and subchondral bone tissues in a rat preclinical model. In conclusion, 3D dECM-based microenvironment-specific biomimetics encapsulated with bioactive exosomes can serve as a novel cell-free recipe for stem cell therapy when treating injured or degenerative joints. This strategy provides a promising platform for complex zonal tissue regeneration whilst holding attractive clinical translation potential.

RANKL-dependent osteoclast differentiation and gene expression in bone marrow-derived cells from adult mice is sexually dimorphic

Sex-specific differences in bone integrity and properties are associated with age as well as the number and activity of cells involved in bone remodeling. The aim of this study was to investigate sex-specific differences in adhesion, proliferation, and differentiation of mouse bone marrow derived cells into osteoclasts. The adherent fraction of bone marrow- derived cells from 12-week-old male and female C57BL/6J mice were assessed for their adhesion, proliferation, and receptor activator of nuclear factor κB (RANKL)-induced differentiation into osteoclasts. Female bone marrow derived macrophages (BMDMs) displayed higher adhesion and proliferation ratio upon macrophage colony stimulating factor (M-CSF) (day 0) and M-CSF + RANKL (day 4) treatment, respectively. On the contrary, male BMDMs differentiated more efficiently into osteoclasts upon RANKL-treatment compared to females (day 5). To further understand these sex-specific differences at the gene expression level, BMDMs treated with M-CSF (day 0) and M-CSF + RANKL (day 4), were assessed for their differential expression of genes through RNA sequencing. M-CSF treatment resulted in 1106 differentially expressed genes, while RANKL-treatment gave 473 differentially expressed genes. Integrin, adhesion, and proliferation-associated genes were elevated in the M-CSF-treated female BMDMs. RANKL-treatment further enhanced the expression of the proliferation- associated genes, and of genes associated with inhibition of osteoclast differentiation in the females, while RANK-signaling-associated genes were upregulated in males. In conclusion, BMDM adhesion, proliferation and differentiation into osteoclasts are sex-specific and may be directed by the PI3K-Akt signaling pathway for proliferation, and the colony stimulating factor 1-receptor and the RANKLsignaling pathway for the differentiation.

A pilot study: Nano-hydroxyapatite-PEG/PLA containing low dose rhBMP2 stimulates proliferation and osteogenic differentiation of human bone marrow derived mesenchymal stem cells.

Bone morphogenetic protein 2 (BMP2) can enhance posterolateral spinal fusion (PLSF). The minimum effective dose that may stimulate mesenchymal stem cells however remains unknown. Nano-hydroxyapatite (nHAp) polyethylene glycol (PEG)/polylactic acid (PLA) was combined with recombinant human BMP2 (rhBMP2). We in vitro evaluated proliferation, differentiation, and osteogenic genes of human bone marrow mesenchymal stem cells with 0.5, 1.0, and 3.0 μg/mL rhBMP2 doses in this study. In vitro experimental study was designed to proliferation by a real-time quantitative cell analysis system and the osteogenic differentiation by alkaline phosphatase (ALP) activity and osteogenic marker (Runx2, OPN, and OCN) gene expressions of human derived bone marrow mesenchymal stem cells (hBMMSCs). nHAp was produced by wet chemical process and characterized by Fourier transform infrared spectrophotometer, scanning electron microscopy, and energy-dispersive x-ray spectroscopy. PEG/PLA polymer was produced at a 51:49 molar ratio. 0.5, 1.0, and 3.0 μg/mL rhBMP2 and nHAp was combined with the polymers. hBMMSCs were characterized by multipotency assays and surface markers were assessed by flow cytometer. The hBMMSC-rhBMP2 containing nHAp-PEG/PLA composite interaction was evaluated by transmission electron microscopy. Proliferative effect was evaluated by real-time proliferation analysis, and osteogenic capacity was evaluated by ALP activity assay and qPCR. hBMMSC proliferation in the 0.5 μg/mL rhBMP2 + nHAp-PEG/PLA and the 1.0 μg/mL rhBMP2 + nHAp-PEG/PLA groups were higher compared to control. 1.0 μg/mL rhBMP2 + nHAp-PEG/PLA and 3.0 μg/mL rhBMP2 + nHAp-PEG/PLA containing composites induced ALP activity on days 3 and 10. 0.5 μg/mL rhBMP2 + nHAp-PEG/PLA application stimulated Runx2 and OPN gene expressions. rhBMP2 + nHAp-PEG/PLA composites stimulate hBMMSC proliferation and differentiation. The nHAp-PEG/PLA composite with low dose of rhBMP2 may enhance bone formation in future clinical PLSF applications.

Chronic Injury of Mice Bone Marrow Multipotent Hematopoietic Progenitor Cells Induced by Ionizing Radiation

To explore the chronic injury and its possible mechanism of ionizing radiation on multipotent hematopoietic progenitor cells (MPPs) by determining the related indicators of MPPs in bone marrow of mice post-radiation. Sixteen C57BL/6 adult mice were randomly divided into normal control and irradiation groups, 8 mice in each group. The mice in irradiation group were exposed to 6 Gy X-ray. The proportion of bone marrow MPPs, their apoptosis and proliferation 2 months after irradiation were detected by flow cytometry. Mitochondrial activity and levels of reactive oxygen species (ROS) in each MPPs population were detected by Mitotracker Red and DCFDA probes, and the senescent state of MPPs in the bone marrow was analyzed. Ionizing radiation could reduce the proportion of MPPs in mouse bone marrow. The proportions and numbers of MPP1, MPP3 and MPP4 in the bone marrow were significantly decreased after whole-body irradiation with 6 Gy X-ray (P<0.05). In addition, radiation significantly reduced the colony-forming capacity of MPPs in bone marrow (P<0.05), the proportions of apoptotic cells in the MPP1 and MPP4 cell populations increased significantly in the bone marrow (P<0.05). The activity of mitochondria was significantly reduced in the bone marrow MPP2, MPP3 and MPP4 cell populations compared with that of the control group (P<0.05). It was also found that the radiation could significantly increase the ROS levels of MPPs in bone marrow, and the content of ROS in the MPP2, MPP3 and MPP4 cell population of the bone marrow was significantly increased(P<0.05). The senescent cells ratios of MPP1, MPP3 and MPP4 cells in the bone marrow after irradiation were significantly higher than those in the control group (P<0.05). Ionizing radiation can cause chronic MPPs damage in mice, which is closely associated with persistent oxidative stress, cells apoptosis, and cellular senescence.

Analysis of immune response of stressed male mice during Chlamydia muridarum lung infection

Chlamydia lung infection caused by Chlamydia trachomatis is a serious lung infection particularly in infants but there are a few studies about lung infection. Chlamydial genital and lung infections in mice is frequently done with Chlamydia muridarum. The purpose of our study was to explore the effect of stress in lung chlamydia infection in male mice. We hypothesize that stress changes the level of cytokine production in monocytes during lung infection. As a The result from these studies include counting of live cells, differentiation of proliferation of bone marrow derived monocytes where the result showed that non-stressed dendritic cells (DCs) WT mice and stressed macrophages (MO) WT mice had sufficient live cells to study. No difference in IL-b production of macrophages of stressed and non-stress mice was observed. Beta2-adrenergic receptor (b2-AR agonist (fenoterol) and antagonist (ICI 118 551) treatment resulted in difference in cytokine production the effect of agonists and antagonists. TNF-alpha production was high in stressed mice compared to non-stress mice for MO. LPS stimulated TNF-alpha production in DC but showed no difference in stressed and non-stressed mice. However, TNF-alpha production in macrophages of non-stressed mice was decreased. The production of IFN-g, in Con A- and LPS-treated splenic T cells. In contrast IL-5, IL-10, and IL-23 production was high in T cells of stressed mice. T cells treated with norepinephrine and bet2-adrenergic receptor agonist, Fenoterol. Overall, the production of cytokines in stressed and non-stressed mice shows variation that may have roles in enhancing protection or increased lung infection.

Langerhans cell hystiocytosis mimicking necrotizing ulcerative periodontitis - A case report

Langerhans cell histiocytosis (LCH) is a rare disease characterized by abnormal proliferation of bone marrow derived histiocytes followed by their deposition in tissues elsewhere in the body causing damage. Oral manifestations of LCH can sometimes be the only presenting symptoms or the first sign of a disseminated disease. Ulcerated lesions of oral mucosa are one of the common clinical manifestations and can help in early diagnosis of the disease when presented. Since the clinical features mimics several other diseases of oral and maxillofacial region it can be misdiagnosed easily as other pathologies. This article highlights the clinicopathological features of a case of LCH in an elderly patient who has reported to our outpatient department.

Treatment with Mesenchymal Stem Cell-Derived Nanovesicle-Containing Gelatin Methacryloyl Hydrogels Alleviates Osteoarthritis by Modulating Chondrogenesis and Macrophage Polarization.

Osteoarthritis is a degenerative disorder that can severely affect joints, and new treatment strategies are urgently needed. Administration of mesenchymal stem cell (MSC)-derived exosomes is a promising therapeutic strategy in osteoarthritis treatment. However, the poor yield of exosomes is an obstacle to the use of this modality in the clinic. Herein, a promising strategy is developed to fabricate high-yield exosome-mimicking MSC-derived nanovesicles (MSC-NVs) with enhanced regenerative and anti-inflammatory capabilities. MSC-NVs are prepared using an extrusion approach and are found to increase chondrocyte and human bone marrow MSC differentiation, proliferation, and migration, in addition to inducing M2 macrophage polarization. Furthermore, gelatin methacryloyl (GelMA) hydrogelsloaded with MSC-NVs (GelMA-NVs) are formulated, which exhibitsustained release of MSC-NVs and are shownto be biocompatible with excellent mechanical properties. In a mouse osteoarthritis model constructed by surgical destabilization of the medial meniscus (DMM), GelMA-NVseffectively ameliorate osteoarthritisseverity, reduce the secretion of catabolic factors, and enhance matrix synthesis. Furthermore, GelMA-NVs induce M2 macrophage polarization and inflammatory response inhibition in vivo. The findings demonstrate that GelMA-NVshold promise for osteoarthritis treatment through modulation of chondrogenesis and macrophage polarization.