Marrow Proliferation Research Articles

Non-Invasive Imaging Methodologies for Assessment of Radiation Damage to Bone Marrow and Kidneys from Peptide Receptor Radionuclide Therapy

Background/Aims: Peptide receptor radionuclide therapy (PRRT) is becoming clinical routine for management of neuroendocrine tumours. The number of PRRT cycles is correlated with treatment effect but theoretically limited by off-target radiation damage to kidneys and bone marrow. New imaging biomarkers for assessment of PRRT tissue damage would enable evaluation of novel renal and bone marrow protective agents, as well as personalised PRRT treatment regiments. Methods: Mice treated with [¹⁷⁷Lu]Lu-DOTA-TATE PRRT or vehicle were examined at baseline and following treatment with [¹⁸F]fluorothymidine (FLT) positron emission tomography (PET) and technetium-99m-mercapto-acetyl-tri-glycine ([^99mTc]Tc-Mag3) single-photon emission tomography (SPECT) to assess dynamic changes in bone marrow proliferation and renal function, respectively. Results: Bone marrow proliferation as assessed by [¹⁸F]FLT was decreased 2 days after PRRT treatment, but not vehicle, compared to baseline (target-to-background ratio [TBR_max] baseline:1.69 ± 0.29 vs. TBR_max PRRT: 0.91 ± 0.02, p < 0.01). Renal function as assessed by [^99mTc]Tc-Mag3 SPECT was similarly decreased 2 days following PRRT compared to vehicle (fractional uptake rate [FUR] vehicle: 0.030 ± 0.014 s^–1 vs. FUR PRRT: 0.0051 ± 0.0028 s^–1, p < 0.01). Conclusion: [¹⁸F]FLT PET and [^99mTc]Tc-Mag3 SPECT are promising techniques for assessing bone marrow and renal injury from [¹⁷⁷Lu]Lu-DOTA-TATE PRRT and may potentially improve patient management by allowing evaluation of protective interventions as well as enabling personalised PRRT treatments.

Low-Intensity Pulsed Ultrasound Relieves Mild and Severe Myelosuppression Induced by Cyclophosphamide in Rabbits.

This study aimed to investigate the effect of low-intensity pulsed ultrasound (LIPUS) on cyclophosphamide (CTX)-induced rabbit myelosuppression. Rabbits (n = 90) were randomly divided into a mild myelosuppression group (n = 40), a severe myelosuppression group (n = 40), and a normal control group (group Cu28 ; n = 10). The mild and severe myelosuppression models were established by daily ear vein injection of 15- and 40-mg/kg CTX for 4 continuous days, respectively. Then they were randomly divided into LIPUS groups (Au and Bu ) and control groups (Ac and Bc ). LIPUS was applied once per day for 20 minutes for 7 (Au7 and Bu7 ) and 28 (Au28 and Bu28 ) days. Physical conditions, mortality, blood cell counts, and bone marrow proliferation were calculated. Erythropoietin interleukin 3, and granulocyte-macrophage colony-stimulating factor levels were measured by an enzyme-linked immunosorbent assay. Flow cytometry was used to detect the granulocyte phagocytosis rate. Hematoxylin-eosin staining was performed to analyze changes of skin and muscle. Compared with the control group, LIPUS improved the number of peripheral blood cells (P < .05) and bone marrow nucleated cells and reduced the mortality of rabbits with myelosuppression of different degrees. Long-term treatment for 28 days had no effect on the levels of erythropoietin, interleukin 3, and granulocyte-macrophage colony-stimulating factor and granulocyte phagocytosis (P > .05). The parts of the skin where LIPUS was applied did not show any burning marks, and the muscle tissue in the path of LIPUS acoustic channels showed no obvious pathologic changes. Low-intensity pulsed ultrasound is a safe and effective method to relieve CTX-induced myelosuppression.

Modifying decellularized aortic valve scaffolds with stromal cell-derived factor-1α loaded proteolytically degradable hydrogel for recellularization and remodeling

Decellularized matrix is of great interest as a scaffold for the tissue engineering heart valves due to its naturally three-dimensional structure and bioactive composition. A primary challenge of tissue engineered heart valves based on decellularized matrix is to grow a physiologically appropriate cell population within the leaflet tissue. In this study, a composite scaffold was fabricated by the combination of a porous matrix metalloproteinase (MMP) degradable poly (ethylene glycol) (PEG) hydrogel that were loaded with stromal cell-derived factor-1α (SDF-1α) and a mechanically supportive decellularized porcine aortic valve. Results demonstrated that the modified scaffold enhanced bone marrow mesenchymal stem cells (BMSC) adhesion, viability and proliferation, and promoted BMSC differentiate into valve interstitial-like cells. Furthermore, these modifications lead to enhanced protection of the scaffold from thrombosis. In vivo assessment by rat subdermal model showed the modified scaffold was highly biocompatible with tissue remodeling characterized by promoting mesenchymal stem cells recruitment and facilitating M2 macrophage phenotype polarization. The surface layers of PEG hydrogel not only could provide a niche for cell migration, proliferation and differentiation, but also protect the scaffolds from rapid degeneration, inflammation and calcification. The intermediate layer of decellularized valve could maintain the organization of the scaffold and perform the valve function. The promising results emphasize the potential of our scaffolds to improve recellularization and promote remodeling of implanted decellularized valves. These findings suggest that the SDF-1α loaded MMP degradable PEG hydrogel modification could be an efficient approach to develop functional decellularized heart valve. Statement of significanceA composite scaffold was fabricated by the combination of a porous matrix metalloproteinase (MMP) degradable poly (ethylene glycol) (PEG) hydrogel that were loaded with SDF-1α and a mechanically supportive decellularized porcine aortic valve. The surface layers of PEG hydrogel not only could provide a niche for cell migration, proliferation and differentiation, but also protect the scaffolds from rapid degeneration, inflammation and calcification. The intermediate layer of decellularized valve could maintain the organization of the scaffold and perform the valve function. The promising results emphasize the ability of our scaffolds to improve recellularization and promote remodeling of implanted decellularized valves. This suggests that the extracellular matrix-based valve scaffolds have potential for clinical applications.

Mussel-Inspired Polydopamine Coating: A General Strategy To Enhance Osteogenic Differentiation and Osseointegration for Diverse Implants.

Surface modifications play an important role in endowing implant surface with excellent biocompatibility and bioactivity. Among the bioinspired surface modifications, the mussel-inspired polydopamine (PDA) has aroused great interest of researchers. Herein, we fabricated PDA on diverse implant surfaces, including biopolymer, biometal, and bioceramic. Then the effects of PDA coating on cell responsive behaviors in vitro and bone formation capacity in vivo were evaluated in detail. The results showed that PDA coating was fabricated on diverse samples surface successfully, which could significantly improve the hydrophilicity of different material surfaces. Furthermore, the results indicated that PDA coating exerted direct enhancing on the adhesion, proliferation and osteogenic differentiation of bone marrow derived mesenchymal stromal cells (BMSCs) through FAK and p38 signaling pathways. During the process, the focal adhesion protein expression and osteogenic-related genes expression level (e.g., ALP, BMP2, BSP, and OPN) were considerably upregulated. Most importantly, the in vivo study confirmed that PDA coating remarkably accelerated new bone formation and enhanced osseointegration performance. Our study uncovered the biological responses stimulated by PDA coating to make a better understanding of cell/tissue-PDA interactions and affirmed that PDA, a bioinspired polymer, has great potential as a candidate and functional bioactive coating medium in bone regeneration and orthopedic application.

Dual modulation of crystallinity and macro-/microstructures of 3D printed porous titanium implants to enhance stability and osseointegration.

The macro architecture and micro surface topological morphology of implants play essential roles in bone tissue regeneration. 3D printing technology provides enormous advantages for the rapid fabrication of personalized bone tissue repair implants. This study presents a demonstration of dual-modulation (DM) 3D printed porous titanium implants to enhance stability and osseointegration. Titanium implants with the first level of modulation of macro porous architecture and mechanical properties are obtained using macro architecture design and 3D printing fabrication. The first level of modulation achieved scaffolds with a wide range of compressive strengths (36.76-139.97 MPa) when varying the scaffold macro architectures. In the second level of modulation of surface topological morphology, alkali treatment, heat treatments and electrochemical deposition of hydroxyapatite coating were conducted for further regulating the biological function of implants. DM 3D printed scaffolds significantly promoted bone marrow mesenchyme stem cell adhesion and proliferation, indicating good cytocompatibility. In addition, in vivo osseointegration experiments indicated that the DM scaffolds formed better tissue-materials interfaces. New bone formation rates in DM scaffolds are higher than those in conventional 3D printed scaffolds after 6 months of implantation (58.1% versus 36.1%). These results demonstrate that DM scaffolds could enhance early stability and osseointegration. This study may provide new insights into the design, fabrication and post-processing of 3D printed porous titanium implants for various applications in personalized bone tissue regeneration.

Biofilm Inhibition via Delivery of Novel Methylthioadenosine Nucleosidase Inhibitors from PVA-Tyramine Hydrogels while Supporting Mesenchymal Stromal Cell Viability.

The rise of antibiotic resistance, coupled with increased expectations for mobility in later life, is creating a need for biofilm inhibitors and delivery systems that will reduce surgical implant infection. A limitation of some of these existing delivery approaches is toxicity exhibited toward host cells. Here, we report the application of a novel inhibitor of the enzyme, methylthioadenosine nucleosidase (MTAN), a key enzyme in bacterial metabolic pathways, which include S-adenosylmethionine catabolism and purine nucleotide recycling, in combination with a poly(vinyl alcohol)-tyramine-based (PVA-Tyr) hydrogel delivery system. We demonstrate that a lead MTAN inhibitor, selected from a screened library of 34 candidates, (2S)-2-(4-amino-5H-pyrrolo3,2-dpyrimidin-7-ylmethyl)aminoundecan-1-ol (31), showed a minimum biofilm inhibitory concentration of 2.2 ± 0.4 μM against a clinical staphylococcal species isolated from an infected implant. We observed that extracellular DNA, a key constituent of biofilms, is significantly reduced when treated with 10 μM compound 31, along with a decrease in biofilm thickness. Compound 31 was incorporated into a hydrolytically degradable photo-cross-linked PVA-Tyr hydrogel and the release profile was evaluated by HPLC studies. Compound 31 released from the PVA-hydrogel system significantly reduced biofilm formation (77.2 ± 8.4% biofilm inhibition). Finally, compound 31 released from PVA-Tyr showed no negative impact on human bone marrow stromal cell (MSC) viability, proliferation, or morphology. The results demonstrate the potential utility of MTAN inhibitors in treating infections caused by Gram-positive bacteria, and the development of a nontoxic release system that has potential for tunability for time scale of delivery.

A mouse model for the study of transplanted bone marrow mesenchymal stem cell survival and proliferation in lumbar spinal fusion.

Bone marrow aspirate has been successfully used alongside a variety of grafting materials to clinically augment spinal fusion. However, little is known about the fate of these transplanted cells. Herein, we develop a novel murine model for the in vivo monitoring of implanted bone marrow cells (BMCs) following spinal fusion. A clinical-grade scaffold was implanted into immune-intact mice undergoing spinal fusion with or without freshly isolated BMCs from either transgenic mice which constitutively express the firefly luciferase gene or syngeneic controls. The in vivo survival, distribution and proliferation of these luciferase-expressing cells was monitored via bioluminescence imaging over a period of 8weeks and confirmed via immunohistochemistry. MicroCT imaging was performed 8weeks to assess fusion. Bioluminescence imaging indicated transplanted cell survival and proliferation over the first 2weeks, followed by a decrease in cell numbers, with transplanted cell survival still evident at the end of the study. New bone formation and increased fusion mass volume were observed in mice implanted with cell-seeded scaffolds. By enabling the tracking of transplanted bone marrow-derived cells during spinal fusion in vivo, this mouse model will be integral to developing a deeper understanding of the biological processes underlying spinal fusion in future studies. These slides can be retrieved under Electronic Supplementary Material.

Retracted: Combined therapy using LHRH-PE40 and anti-CD40 dendritic cells substantially eliminate tumor cells.

LHRH-PE40 was used to promote the proliferation of bone marrow derived cell (BMDC) and improve the antigen-presenting ability of BMDC as well as the immune function via the CD40 signal pathway. LHRH-PE40 was also implicated in cancer treatment, targeting a variety of cancer cells that express luteinizing hormone-releasing hormonereceptor (LHRHR). In the present study, the mechanism and efficacy of LHRH-PE40 were addressed in the following three aspects. Enzyme-linked immunosorbent assay was performed to confirm the binding specificity of LHRH-PE40 to LHRHR. The killing effect of LHRH-PE40 on target cells was mediated by LHRHR, which specifically killed LHRHR-positive target cells while the minimal cytotoxicity of LHRHR-negative cells is negligible. Spiegelmers, a molecule mutually exclusive with GnRH and developed by Sven Klussmann and Dr Sven Klussmann of NOXXON Pharmaceuticals in Germany, demonstrated that LHRH-PE40 maintains a combinatory characteristics of LHRH and LHRHR. In the end, the mechanism of LHRH-PE40 underlying induction of apoptosis at low concentration and prolonged conditions was firstly demonstrated by the basic method of detecting apoptosis to induce apoptosis. It provided a scientific basis for clinical application of LHRH-PE40 and laid a foundation for the further study of LHRH-PE40 on inducing apoptosis of target cells. The target cells herein refer to tumor cells that overexpress LHRHR. This study shows that activated DC can more effectively promote the proliferation of CD4+ T cells, and initially proved that DC carrying anti-CD40 antibody promoted the immune treatment of the tumor. Combining LHRH-PE40 with anti-CD40 DCs achieved substantially improved efficacy in killing tumor cells.

Mesenchymal stem cells gene signature in high-risk myeloma bone marrow linked to suppression of distinct IGFBP2-expressing small adipocytes.

Recent studies suggest that multiple myeloma (MM) induces proliferation and expansion of bone marrow (BM) mesenchymal stem cells (MSCs), but others showed that MM cells induce MSC senescence. To clarify the interaction between MM and MSCs, we exploited our established MSC gene signature to identify gene expression changes in myeloma MSCs and associated functional differences. Single MSCs from patients with MM had changes in expression of genes associated with cellular proliferation and senescence and a higher proportion of senescent cells and lower proliferative potential than those from age-matched healthy donors. Single MSCs from both sources heterogeneously express MSC genes associated with adipogenesis and osteoblastogenesis. We identified the gene encoding insulin-like growth factor-binding protein 2 (IGFBP2), an MSC gene commonly altered in high risk MM, as under-expressed. Morphologically, IGFBP2+ cells are underrepresented in MM BM compared to smouldering MM. Strong IGFBP2 and adiponectin co-expression was detected in a subset of small adipocytes. Co-culturing normal MSCs with myeloma cells suppressed MSC differentiation to adipocytes and osteoblasts, and reduced expression of IGFBP2 and adiponectin. Recombinant IGFBP2 blocked IGF1-mediated myeloma cell growth. Our data demonstrate that myeloma MSCs are less proliferative and that IGFBP2+ small adipocytes are a distinct mesenchymal cell population suppressed by myeloma.

Immunomodulatory effects of rhesus monkey bone marrow-derived mesenchymal stem cells in serum-free conditions

Mesenchymal stem cells (MSCs) have generated tremendous interest for treating various diseases due to their self-renewal and differentiation capacities. Many studies have demonstrated the immunoregulatory capability of MSCs; however, most of these studies were conducted with fetal bovine serum (FBS), which has an uncertain composition. In this study, we established a serum-free, xeno-free, completely chemically defined medium for the proliferation and expansion of rhesus monkey bone marrow (BM)-derived MSCs (rBMSCs) in vitro. The growth kinetics, characteristics, immunophenotype, and immunosuppressive abilities of rBMSCs grown in serum-free media (SFM) were evaluated and compared with those of cells grown in serum-containing media (SCM). Moreover, we employed RNA sequencing to evaluate the expression pattern of genes related to immune responses in both culture conditions. Compared to cells grown in SCM, rBMSCs grown in SFM exhibited better biological characteristics regarding cell proliferation and immunosuppressive abilities. Cells from both media types exhibited similar immunophenotypic expression patterns for CD29, CD34, CD45, HLA-DR, CD73, CD90, and CD105. Gene Ontology (GO) terms, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and Gene Set Enrichment Analysis (GSEA) revealed that CXCL8 was downregulated by 4.1 fold in SFM-cultured rBMSCs compared with those in SCM. Furthermore, the mixed lymphocyte culture revealed that the proliferation activity and the expression levels of inflammatory factors of peripheral blood mononuclear cells (PBMCs) were significantly decreased after the addition of the CXCL8 neutralizing antibody, which was related to the elevated immunosuppressive abilities of SFM-suspended rBMSCs. These results suggest a possible cell culture method as well as immunoregulatory mechanisms for clinical cell therapies requiring nonanimal-derived components.

Zirconium modified calcium‐silicate‐based nanoceramics: An in vivo evaluation in a rabbit tibial defect model

AbstractOver the past two decades calcium silicate (Ca–Si) based ceramics have been introduced as bioactive materials for bone tissue engineering applications and repairing bone defects. Recently, it is reported that zirconium modified calcium‐silicate‐based (Ca3ZrSi2O9, Baghdadite) ceramics stimulate cell proliferation and osteogenic differentiation of osteoblasts. The aim of this work was preparation, characterization, and evaluation of biological properties of Baghdadite in the nanoparticles form. These nanoparticles were synthesized by sol‐gel method and were annealed at 1150°C. Fabricated nanoparticles were investigated using XRD, FE‐SEM, EDX, and MTT in vitro, then implanted to the rabbit tibia as a bone animal model. The characterization results showed that nanoparticles with expected composition were successfully synthesized using sol‐gel method. According to MTT result, these nanoparticles were nontoxic. In addition, the proliferation of bone marrow derived mesenchymal stem cells was increased after 96 hours of culturing in the presence of nanoparticles compared to the control. Histopathological results also demonstrated that the defected bone was completely regenerated when implanted with nanoparticles after 6 weeks. Taking together, according to the results, Ca3ZrSi2O9 nanoparticles as a bioceramic with excellent biological properties can be applied for improvement of bone lesions healing process in the orthopedic and dental application.

Characteristics of bone marrow cells in 107 patients with juvenile idiopathic arthritis: A retrospective study.

Few studies to date have reported on the myelodysplastic features of children with juvenile idiopathic arthritis (JIA). Bone marrow specimens were collected from 107 patients aged from 7-12 years who were initially diagnosed with JIA between May 2013 and October 2015. In 107 patients with JIA, bone marrow proliferation was higher than normal and hemophagocytes were more easily observed than usual. The characteristics of bone marrow cells in 107 patients with JIA were investigated and the associations of these characteristics with the disease was discussed in the present study. It was noticed that there were similar changes in the myeloid, erythropoietic and megakaryopoietic series in the majority of bone marrow specimens; the presence of hemophagocytes was also reported. The present findings suggest that JIA is associated with specific myelodysplastic changes, and that cellular immune system dysfunction and overreactive inflammatory cytokines may contribute to the development of these myelodysplastic changes in the bone marrow.

Comparative effect of melatonin and quercetin in counteracting LPS induced oxidative stress in bone marrow mononuclear cells and spleen of Funambulus pennanti

A majority of cellular diseases, independent of their origin, are characterized by a dramatic increase in Reactive Oxygen Species (ROS) in response to stress. In most cases, the uncontrolled detrimental ROS outburst is difficult to handle for the cellular machinery and eventually leads to cell mortality. In this study, we compare the antioxidant efficacy of quercetin and melatonin to find out a better alternative against lipopolysaccharide (LPS) induced tissue injury by oxidative stress in Funambulus pennanti. Transient exposure to LPS significantly increased ROS generation and lipid peroxidation levels in bone marrow mononuclear cells (MNCs) and spleen which was further corroborated by decreased activities of SOD, CAT and Gpx enzymes. It also downregulate the expression of cellular oxidative stress response proteins Nrf-2 and HO-1 in spleen and decreases the proliferation of bone marrow derived Granulocyte macrophage-colony forming unit cells (GM-CFU). Both melatonin and quercetin pre-treatments rescued these effects, however, our results indicated that the efficacy of melatonin to overcome oxidative stress was significantly better than quercetin. Our findings support the idea that melatonin is a better antioxidant and immunomodulator as compared to other alternatives and perhaps may be employed in the development of effective therapeutics against ROS dominated diseases.

Silk fibroin/nanohydroxyapatite hydrogels for promoted bioactivity and osteoblastic proliferation and differentiation of human bone marrow stromal cells

Silk fibroin (SF) is a natural, biocompatible, and biodegradable polymer having a great potential for the successful regeneration of damaged bone tissue. In the present work, nanohydroxyapatite (nanoHA) was incorporated into SF polymer to form a bioactive composite hydrogel for applications as bone implants. The degradation and bioactive properties of SF/nanoHA composite hydrogels were evaluated. Additionally, biological investigations of human bone marrow stromal cells (hBMSCs) viability, proliferation and differentiation to the osteoblastic phenotype were conducted. The incorporation of nanoHA in SF polymer matrices improved the bioactivity of the hydrogels. The biological results highlighted that the SF/nanoHA composite hydrogels are suitable for hBMSCs attachment and proliferation, while a test for alkaline phosphatase (ALP) and bone morphogenetic protein 2 (BMP-2) expression suggested osteoblast differentiation. Additionally, a cell staining method for ALP allowed to observe cell infiltration with active production of ALP by the infiltrated cells, paving the way to use the proposed composite hydrogel for bone tissue regeneration.

The p38-MK2/3 Module Is Critical for IL-33-Induced Signaling and Cytokine Production in Dendritic Cells.

IL-33 is an IL-1 cytokine superfamily member. Binding of IL-33 to the IL-33R induces activation of the canonical NF-κB signaling and activation of MAPKs. In bone marrow-derived dendritic cells, IL-33 induces the production of IL-6, IL-13, and TNF-α. However, the signaling pathways resulting in IL-33-induced effector functions of dendritic cells are unknown. In this article, we show that the IL-33-induced cytokine production is only partly dependent on p65. Thereby, p65 mediates the production of IL-6, but not of IL-13, whereas the p38-Mapk-activated protein kinases 2/3 (MK2/3) signaling module mediates the IL-13, but not the IL-6, production. In addition, GM-CSF, which is critical for the differentiation and proliferation of bone marrow-derived dendritic cells, potentiates the p65-dependent IL-6 and the p38-MK2/3-dependent IL-13 production. Furthermore, we found that effective TNF-α production is only induced in the presence of GM-CSF and IL-33 via the p38-MK2/3 signaling module. Taken together, we found that the p38-MK2/3 signaling module is essential to mediate IL-33-induced cytokine production in dendritic cells.

The small GTPase RhoU lays downstream of JAK/STAT signaling and mediates cell migration in multiple myeloma

Multiple myeloma is a post-germinal center B-cell neoplasm, characterized by the proliferation of malignant bone marrow plasma cells, whose survival and proliferation is sustained by growth factors and cytokines present in the bone marrow microenvironment. Among them, IL-6 triggers the signal downstream of its receptor, leading to the activation of the JAK/STAT pathway. The atypical GTPase RhoU lays downstream of STAT3 transcription factor and could be responsible for mediating its effects on cytoskeleton dynamics. Here we demonstrate that RHOU is heterogeneously expressed in primary multiple myeloma cells and significantly modulated with disease progression. At the mRNA level, RHOU expression in myeloma patients correlated with the expression of STAT3 and its targets MIR21 and SOCS3. Also, IL-6 stimulation of human myeloma cell lines up-regulated RHOU through STAT3 activation. On the other hand, RhoU silencing led to a decrease in cell migration with the accumulation of actin stress fibers, together with a decrease in cyclin D2 expression and in cell cycle progression. Furthermore, we found that even though lenalidomide positively regulated RhoU expression leading to higher cell migration rates, it actually led to cell cycle arrest probably through a p21 dependent mechanism. Lenalidomide treatment in combination with RhoU silencing determined a loss of cytoskeletal organization inhibiting cell migration, and a further increase in the percentage of cells in a resting phase. These results unravel a role for RhoU not only in regulating the migratory features of malignant plasma cells, but also in controlling cell cycle progression.

Evaluation of Biocompatibility and Osteogenic Potential of Tricalcium Silicate–based Cements Using Human Bone Marrow–derived Mesenchymal Stem Cells

IntroductionThe success of endodontic regeneration lies in the appropriate combination of stem cells and bioactive materials. Several novel dental materials are available on the market in this regard. Hence, the current study aimed to evaluate the proliferation, differentiation, and osteogenic potential of human bone marrow–derived mesenchymal stem cells (hBMSCs) onto biomaterials like ProRoot MTA (MTA; Dentsply Tulsa Dental, Tulsa, OK), Biodentine (BD; Septodont, Saint Maur de Fosses, France), and EndoSequence Root Repair Material (ERRM; Brasseler USA, Savannah, GA). MethodsDental cements were formulated into discs and assessed for their biocompatibility. hBMSCs were used to study biocompatitibility and the proliferative and osteogenic potential of these dental cements. A live dead assay was performed using confocal microscopy to study the biocompatibility, proliferation, and cell attachment property of the cements. An 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was also performed on days 1, 3, 5, and 7 to study growth kinetics. The osteogenic potential of these cements was studied by inducing hBMSCs over them using osteogenic differentiation medium (assessed by alkaline phosphatase assay). ResultsERRM and MTA have shown the best biocompatibility among the tricalcium silicate materials used with no significant difference between them. Both have shown significantly higher osteogenic bioactivity than BD. All 3 tricalcium silicate cements support good adherence of hBMSCs. ConclusionsAll of the dental cements used in this study are biocompatible with the potential to induce proliferation and osteogenic differentiation of hBMSCs. Therefore, the newly introduced ERRM can be the material of choice in various endodontic applications.

Bioactive Silk Hydrogels with Tunable Mechanical Properties.

Developing bioactive hydrogels with potential to guide the differentiation behavior of stem cells has become increasingly important in the biomaterials field. Here, silk hydrogels with tunable mechanical properties were developed by introducing inert silk fibroin nanofibers (SNF) within an enzyme crosslinked system of regenerated silk fibroin (RSF). After the crosslinking reaction of RSF, the inert SNF was embedded into the RSF hydrogel matrix, resulting in improved mechanical properties. Tunable stiffness in the range of 9-60 KPa was achieved by adjusting the amount of the added NSF, significantly higher than SNF-free hydrogels formed under same conditions (about 1 KPa). In addition, the proliferation of rat bone marrow derived mesenchymal stem cells cultured on the composite hydrogels and differentiated into endothelial cells, myoblast and osteoblast cells was improved, putatively due to the control of stiffness of the hydrogels. Bioactive and tunable silk-based hydrogels were prepared via a composite SNF and crosslinked RSF system, providing a new strategy to design silk biomaterials with tunable mechanical and biological performance.

Eugenol enhances proliferation and migration of mouse bone marrow-derived mesenchymal stem cells in vitro

Mesenchymal stem cells (MSCs) have received considerable attention in regenerative medicine during the past decade. Eugenol is a natural and versatile vegetable molecule, which has a wide variety of therapeutic effects. Although different biological and pharmaceutical functions of Eugenol are well known, its effect on MSCs has not been studied yet. Therefore, this study was focused on investigating the effect of Eugenol on the proliferation and migration of bone marrow (BM)-derived MSCs in vitro. To do so, BM-MSCs were isolated from 4 to 8 weeks old NMRI mice. Cytotoxicity of Eugenol on MSCs was evaluated by MTT assay at 24, 48 and 72 h after treatment. In addition, its effect was assessed on the proliferation and migration of MSCs using wound healing assay in vitro and quantitative gene expression analysis for Oct4, Sox2, Cyclin-D1, Rex1, Tex10, Cxcr4, Vla4 and c-Met. Results showed that Eugenol reduced the number of MSCs in a dose- and time-dependent manner. The median inhibition concentration of Eugenol on MSCs was 400 μg/ml at 24 and 48 h and 200 μg/ml at 72 h after treatment. Moreover, about 90% viability of MSCs was detected at concentrations ≤12.5 μg/ml. The wound healing assay and gene expression analysis demonstrated that Eugenol promoted the migratory potential of MSCs through up-regulation of c-Met. Moreover, Eugenol has enhanced the proliferation of MSCs via over-expression of Sox2, Rex1 and Tex10. In conclusion, this study revealed that Eugenol enhances the proliferation and migration of MSCs, and thus this will be beneficial to the field of regenerative medicine.

Effect of Autologous and Allogeneic Platelet Rich Plasma on Viability, Proliferation and Chondrogenesis of Equine Bone Marrow Derived Mesenchymal Stromal Cells

Effect of Autologous and Allogeneic Platelet Rich Plasma on Viability, Proliferation and Chondrogenesis of Equine Bone Marrow Derived Mesenchymal Stromal Cells