11 research outputs found
RNA sequencing suggests that non-coding RNAs play a role in the development of acquired haemophilia
Funding Information: Adrian Bogdan Tigu and Ionut Hotea contributed equally to the current manuscript and are both considered first author. The authors also gratefully acknowledge the support of Sergiu Pasca, M.D. – Johns Hopkins University School of Medicine, Baltimore, United States, for his contribution on the statistical analysis. Funding Information: IH is funded by an internal grant of the Iuliu Hatieganu University – School of Doctoral Studies. BT is supported by a national grant of the Romanian Academy of Scientists (Academia Oamenilor de Stiinta din Romania) 2023–2024. ABT, DG, JTB and VG are supported by an international collaborative grant of the European Economic Space between Romania and Iceland 2021–2023: ‘Cooperation strategy for knowledge transfer, internationalization and curricula innovation in the field of research education at the 3rd level of study –AURORA.’. The experiments were funded by an international grant awarded by the Novo Nordisk Haemophilia Foundation to the Romanian Haematology Society—Romania 4. CT is supported by a grant by grants awarded by the Romanian National Ministry of 350 Research, Innovation, and Digitalisation: Project PN‐III‐P4‐ID‐PCE‐2020‐1118. Publisher Copyright: © 2023 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd. © 2023 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.Acquired haemophilia (AH) is a rare disorder characterized by bleeding in patients with no personal or family history of coagulation/clotting-related diseases. This disease occurs when the immune system, by mistake, generates autoantibodies that target FVIII, causing bleeding. Small RNAs from plasma collected from AH patients (n = 2), mild classical haemophilia (n = 3), severe classical haemophilia (n = 3) and healthy donors (n = 2), for sequencing by Illumina, NextSeq500. Based on bioinformatic analysis, AH patients were compared to all experimental groups and a significant number of altered transcripts were identified with one transcript being modified compared to all groups at fold change level. The Venn diagram shows that haemoglobin subunit alpha 1 was highlighted to be the common upregulated transcript in AH compared to classical haemophilia and healthy patients. Non-coding RNAs might play a role in AH pathogenesis; however, due to the rarity of HA, the current study needs to be translated on a larger number of AH samples and classical haemophilia samples to generate more solid data that can confirm our findings.Peer reviewe
BMP-2 Delivery through Liposomes in Bone Regeneration
Bone regeneration is a central focus of maxillofacial research, especially when dealing with dental implants or critical sized wound sites. While bone has great regeneration potential, exogenous delivery of growth factors can greatly enhance the speed, duration, and quality of osseointegration, making a difference in a patient’s quality of life. Bone morphogenic protein 2 (BMP-2) is a highly potent growth factor that acts as a recruiting molecule for mesenchymal stromal cells, induces a rapid differentiation of them into osteoblasts, while also maintaining their viability. Currently, the literature data shows that the liposomal direct delivery or transfection of plasmids containing BMP-2 at the bone wound site often results in the overexpression of osteogenic markers and result in enhanced mineralization with formation of new bone matrix. We reviewed the literature on the scientific data regarding BMP-2 delivery with the help of liposomes. This may provide the ground for a future new bone regeneration strategy with real chances of reaching clinical practice
Extramedullary Hematopoiesis of the Liver and Spleen
Hematopoiesis is the formation of blood cellular components and, consequently, immune cells. In a more complete definition, this process refers to the formation, growth, maturation, and specialization of blood cells, from the hematopoietic stem cell, through the hematopoietic progenitor cells, to the s pecialized blood cells. This process is tightly regulated by several elements of the bone marrow microenvironment, such as growth factors, transcription factors, and cytokines. During embryonic and fetal development, hematopoiesis takes place in different organs: the yolk sac, the aorta–gonad mesonephros region, the lymph nodes, and not lastly, the fetal liver and the spleen. In the current review, we describe extramedullary hematopoiesis of the spleen and liver, with an emphasis on myeloproliferative conditions
Gradual Drug Release Membranes and Films Used for the Treatment of Periodontal Disease
Periodontitis is an inflammatory disease that, if not treated, can cause a lot of harm to the oral cavity, to the patients’ quality of life, and to the entire community. There is no predictable standardized treatment for periodontitis, but there have been many attempts, using antibiotics, tissue regeneration techniques, dental scaling, or root planning. Due to the limits of the above-mentioned treatment, the future seems to be local drug delivery systems, which could gradually release antibiotics and tissue regeneration inducers at the same time. Local gradual release of antibiotics proved to be more efficient than systemic administration. In this review, we have made a literature search to identify the articles related to this topic and to find out which carriers have been tested for drug release as an adjuvant in the treatment of periodontitis. Considering the inclusion and exclusion criteria, 12 articles were chosen to be part of this review. The selected articles indicated that the drug-releasing carriers in periodontitis treatment were membranes and films fabricated from different types of materials and through various methods. Some of the drugs released by the films and membranes in the selected articles include doxycycline, tetracycline, metronidazole, levofloxacin, and minocycline, all used with good outcome regarding their bactericide effect; BMP-2, Zinc–hydroxyapatite nanoparticles with regenerative effect. The conclusion derived from the selected studies was that gradual drug release in the periodontal pockets is a promising strategy as an adjuvant for the treatment of periodontal disease
Cannabidiol and Vitamin D3 Impact on Osteogenic Differentiation of Human Dental Mesenchymal Stem Cells
Background and objective: The aim of the present study was to establish a new differentiation protocol using cannabidiol (CBD) and vitamin D3 (Vit. D3) for a better and faster osteogenic differentiation of dental tissue derived mesenchymal stem cells (MSCs). Materials and methods: MSCs were harvested from dental follicle (DFSCs), dental pulp (DPSCs), and apical papilla (APSCs) of an impacted third molar of a 17-year old patient. The stem cells were isolated and characterized using flow cytometry; reverse transcription polymerase chain reaction (RT-PCR); and osteogenic, chondrogenic, and adipogenic differentiation. The effects of CBD and Vit. D3 on osteogenic differentiation of dental-derived stem cell were evaluated in terms of viability/metabolic activity by alamar test, expression of collagen1A, osteopontin (OP), osteocalcin (OC), and osteonectin genes and by quantification of calcium deposits by alizarin red assay. Results: Stem cell characterization revealed more typical stemness characteristics for DFSCs and DPSCs and atypical morphology and markers expression for APSCs, a phenotype that was confirmed by differences in multipotential ability. The RT-PCR quantification of bone matrix proteins expression revealed a different behavior for each cell type, APSCs having the best response for CBD. DPSCs showed the best osteogenic potential when treated with Vit. D3. Cultivation of DFSC in standard stem cell conditions induced the highest expression of osteogenic genes, suggesting the spontaneous differentiation capacity of these cells. Regarding mineralization, alizarin red assay indicated that DFSCs and APSCs were the most responsive to low doses of CBD and Vit. D3. DPSCs had the lowest mineralization levels, with a slightly better response to Vit. D3. Conclusions: This study provides evidence that DFSCs, DPSCs, and APSCs respond differently to osteoinduction stimuli and that CBD and Vit. D3 can enhance osteogenic differentiation of these types of cells under certain conditions and doses
Comparative in vitro study regarding the biocompatibility of titanium-base composites infiltrated with hydroxyapatite or silicatitanate
Additional file 5: Figure S1. of Dental follicle stem cells in bone regeneration on titanium implants
Neuronal differentiation of Df stem cells. Protocol of neuronal differentiation. DF stem cells were seeded in 6 well plates at cell density of 20 × 105 cells/well. When cells reached confluence a two steps protocol was applied: cells were cultivated for 48 h in presence of neuronal differentiation medium 1 consisting of DMEM high glucose/F12-HAM (1:1 ratio), 10 % fetal bovine serum (FBS), 1 % antibiotics,2 mM glutamine, 1 % non-essential aminoacids (NEA), supplemented with 10 ng/ml Epidermal Growth Factor (EGF), 10 ng/ml basic Fibroblast Growth Factor (bFGF), 2 % B27 and 1 % N2 Supplement. Afterwards cells were exposed to the differentiation medium 2 for 3 weeks: DMEM high glucose/F12-HAM, 10 % FBS, 1 % antibiotics, 2 mM glutamine, 1 % NEA, 1 % N2 Supplement, 2 % B27 Supplement, 3 μM all-trans retinoic acid, 0.5 mM 3-isobutyl-1-methyl-xanthine (IBMX) (all reagents were purchased from Sigma Aldrich). At the end of 3 weeks of DF stem cultivation with neuronal differentiation medium, cells were fixed and immunocytochemical stained for neurofilaments and CD 133 expression. As shown in Figure S1, the stained cells expressed positivity only for neurofilaments. Figure S1: Fluorescence image of DF stem cells induced to differentiate into neuronal cells. Cells were stained with anti neurofilaments antibody conjugated with FITC green), anti CD 133 conjugated with Texas red (red) and nuclei were counterstained with DAPI (magnification ×100). (PNG 451 kb
Additional file 6: Figure S6. of Dental follicle stem cells in bone regeneration on titanium implants
The physical chemical characterization of titanium coatings. For proving that by used sol–gel method we obtain these nanocrystalline forms of HA and anatase, after heat treatments at quite low temperatures. Figure S6: XRD pattern of hydroxyapatite sample after 600 oC heat treatment. (PNG 7 kb
Additional file 3: Figure S3. of Dental follicle stem cells in bone regeneration on titanium implants
Alamar Blue viability assay. For testing the viability and proliferation rate of DF stem cells cultivated on titanium implants, cells seeded at a cell density of 1.2 × 105 cells/well in 12-wells plates were stained with Alamar blue solution at different periods of time (24 h, 4 and 12 days). Briefly, 100 μl of Alamar blue solution (Invitrogen) was added in each well containing 900 μl stem cell medium or differentiation medium (OS and OC). Each sample was evaluated in triplicate. After 1 h of incubation in dark at 37 °C, the medium was transferred to another 12-well plates and the absorbance was read using a BioTek Synergy 2 plate reader at 570 nm (Winooski, VT, USA). Statistical analysis was performed using t test and two-way ANOVA, Bonferroni posttest. Results: No important differences were observed between titanium implants in terms of cell viability. Statistical differences were noticed only for the 24 h culture between cell cultured on control titanium implants (Ti ctrl) and implants infiltrated with HA (Ti HA) (Figure S3). Two-way ANOVA statistical analysis revealed differences regarding the time factor (24 h vs. 12 days and 4 vs. 12 days) Figure S3: Graphical aspect of optical density values (absorbance at 570 nm) of Alamar blue staining of DF stem cells cultivated with standard stem cell medium evaluated after 24 h, 4 and 12 days (Legend: TiCtrl- Ti6Al7Nb alloy porous titanium, TiHA-titanium infiltrated with hydroxyapatite, TiSiO2-titanium infiltrated with silicatitanate). (PNG 1002 kb