Treatment of gingival fibroblasts with PDL extracellular vesicles results in promotion of Wnt signalling pathway and osteogenic differentiation. PDL secretome shows selective wound healing and matrix remodelling which can have implications for future periodontal regenerative strategies.
- MeSH
- Cell Differentiation MeSH
- Extracellular Vesicles * physiology MeSH
- Fibroblasts physiology MeSH
- Gingiva cytology MeSH
- Wound Healing physiology MeSH
- Humans MeSH
- Osteogenesis physiology MeSH
- Periodontal Ligament * cytology physiology MeSH
- Regeneration * physiology MeSH
- Wnt Signaling Pathway physiology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Extracellular vesicles can play an important role in the processes occurring after stem cell transplantation, preventing cell apoptosis, stimulating immunological processes, and promoting the synthesis of extracellular matrix. Human follicular fluid (FF) can be a source of a subpopulation of cells with mesenchymal stem cells (MSCs) properties. Moreover these subpopulations of FF cells can differentiate into osteoblasts. In presented studies flow cytometry of ovarian FF cells confirmed positive expression of MSCs markers such as: CD44, CD90, CD105, CD73 and negative expression of a hematopoietic marker: CD45. The CD90+, CD105+, CD45- cell subpopulation has been obtained during magnetic separation using appropriate antibodies conjugated with microbeads. The extracellular vesicles (EVs) secreted by the cells during osteodifferentiation process differed from those secreted by cells culture in the basal medium. Based on the previous and current electron microscopy research, changes in size, number, and shape would support the notion that released EVs could be crucial to the ovarian FF cell subpopulation differentiation process. Osteogenic differentiation has been confirmed via Alizarin red staining. Therefore, follicular fluid (FF) can be a new source of a cell subpopulation with MSC properties, with the cells capable of differentiating into the osteogenic lineage. EVs could play a key role as mediators in tissue regeneration, especially bone tissue regeneration.
- MeSH
- Cell Differentiation * MeSH
- Extracellular Vesicles * ultrastructure metabolism MeSH
- Follicular Fluid * cytology metabolism MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Mesenchymal Stem Cells * cytology metabolism MeSH
- Osteoblasts cytology metabolism MeSH
- Osteogenesis * MeSH
- Flow Cytometry MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
The etiology of bone loss in celiac disease (CeD) remains a clinical challenge, with uncertainties present such as the extent of involvement of malabsorption and inflammation-induced osteoresorption processes in development of osteopenia/osteoporosis (OPN/OP), or reasons for failure to achieve healthy bone mass (BMD) even after long-term gluten-free diet (GFD) treatment. This observational prospective study explores the in vitro osteoclastogenic potential of peripheral blood precursors originating from adult active (newly diagnosed and untreated) celiac disease patients (aCeD) and describes the longitudinal changes in osteoclastogenesis after long-term adherence to GFD. To find connections between in vitro observations and in vivo bone metabolism changes, serum levels of 25(OH)D3, PTH, bCTX, PINP, CRP, IL-6, RANKL and OPG were measured before and after GFD and levels of these markers were correlated with the rate of osteoclastogenesis in vitro. OPG and IL-6 showed associations with BMD and/or presence of OPN/OP. Patients after GFD (CeD-GFD) exhibited improved BMD and increased serum 25(OH)D3 levels, alongside reduced bCTX and PINP levels. Compared to healthy donors, aCeD osteoclast genesis in vitro was higher and, surprisingly, remained elevated even in CeD-GFD patients. Negative correlation was found between osteoclastogenesis rate and serum OPG in aCeD, while osteoclastogenesis rate positively correlated with PTH in CeD-GFD. These results highlight OPG as marker for risk of OPN/OP in CeD and suggest that improvement of BMD after GFD is a result of uncoupling between bone metabolism and osteoresorptive action of osteoclasts after GFD.
- MeSH
- Diet, Gluten-Free * MeSH
- Celiac Disease * diet therapy metabolism MeSH
- Adult MeSH
- Interleukin-6 * blood metabolism MeSH
- Bone Density MeSH
- Middle Aged MeSH
- Humans MeSH
- Osteogenesis MeSH
- Osteoclasts metabolism MeSH
- Osteoporosis etiology metabolism MeSH
- Osteoprotegerin * blood metabolism MeSH
- Prospective Studies MeSH
- Vitamin D blood administration & dosage MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Observational Study MeSH
OBJECTIVE: Insulin-sensitizing drugs, despite their broad use against type 2 diabetes, can adversely affect bone health, and the mechanisms underlying these side effects remain largely unclear. Here, we investigated the different metabolic effects of a series of thiazolidinediones, including rosiglitazone, pioglitazone, and the second-generation compound MSDC-0602K, on human mesenchymal stem cells (MSCs). METHODS: We developed 13C subcellular metabolomic tracer analysis measuring separate mitochondrial and cytosolic metabolite pools, lipidomic network-based isotopologue models, and bioorthogonal click chemistry, to demonstrate that MSDC-0602K differentially affected bone marrow-derived MSCs (BM-MSCs) and adipose tissue-derived MSCs (AT-MSCs). In BM-MSCs, MSDC-0602K promoted osteoblastic differentiation and suppressed adipogenesis. This effect was clearly distinct from that of the earlier drugs and that on AT-MSCs. RESULTS: Fluxomic data reveal unexpected differences between this drug's effect on MSCs and provide mechanistic insight into the pharmacologic inhibition of mitochondrial pyruvate carrier 1 (MPC). Our study demonstrates that MSDC-0602K retains the capacity to inhibit MPC, akin to rosiglitazone but unlike pioglitazone, enabling the utilization of alternative metabolic pathways. Notably, MSDC-0602K exhibits a limited lipogenic potential compared to both rosiglitazone and pioglitazone, each of which employs a distinct lipogenic strategy. CONCLUSIONS: These findings indicate that the new-generation drugs do not compromise bone structure, offering a safer alternative for treating insulin resistance. Moreover, these results highlight the ability of cell compartment-specific metabolite labeling by click reactions and tracer metabolomics analysis of complex lipids to discover molecular mechanisms within the intersection of carbohydrate and lipid metabolism.
- MeSH
- Adipogenesis * drug effects MeSH
- Cell Differentiation drug effects MeSH
- Hypoglycemic Agents pharmacology MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Metabolomics MeSH
- Mesenchymal Stem Cells * drug effects metabolism MeSH
- Osteogenesis * drug effects MeSH
- Pioglitazone pharmacology MeSH
- Rosiglitazone pharmacology MeSH
- Thiazolidinediones * pharmacology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
INTRODUCTION: The primary aim of this study was to assess the amount and long-term stability of orthodontically created bone in patients with agenesis of maxillary lateral incisors after canine distalization. The secondary aim was to explore the impact of patient age on the process of alveolar bone resorption. METHODS: A group of patients with agenesis of the maxillary permanent lateral incisor was examined at 4 time points: the beginning of orthodontic treatment (T1, n = 80), the end of treatment (T2, n = 80), 2-5 years after treatment (T3, n = 79), and 12-15 years after treatment (T4, n = 32). The width of the edentulous alveolar bone was measured from study casts at the level of the bone ridge (point A) and 5 mm apically from the alveolar ridge (point B). Alveolar ridge height was also recorded using panoramic radiographs at all time points. Paired t tests, 2-sample t tests, Friedman test with Bonferroni correction, Spearman`s correlation, and linear regression tests were used to analyze the data. RESULTS: The alveolar ridge width was reduced by an average of 0.44 mm at point A and 0.47 mm at point B during the 12-15 years after treatment (T2-T4) and by 0.21 mm and 0.19 mm during the last 10 years (T3-T4). The alveolar ridge height was reduced by 0.59 mm between T2 and T4 and by 0.05 mm between T3 and T4. All reductions in ridge width and height were statistically significant (P <0.05). However, no significant correlation was observed between patient age and changes in alveolar bone parameters (P >0.05). CONCLUSIONS: Although the reductions in alveolar ridge dimensions were statistically significant, the orthodontically created bone after canine distalization remained stable 12-15 years after treatment in both the horizontal and vertical dimensions. Patient age did not significantly influence alveolar bone changes.
- MeSH
- Anodontia * therapy diagnostic imaging MeSH
- Jaw, Edentulous MeSH
- Child MeSH
- Adult MeSH
- Humans MeSH
- Maxilla * MeSH
- Adolescent MeSH
- Young Adult MeSH
- Follow-Up Studies MeSH
- Osteogenesis physiology MeSH
- Tooth Movement Techniques * methods MeSH
- Alveolar Process * diagnostic imaging pathology abnormalities MeSH
- Radiography, Panoramic MeSH
- Alveolar Bone Loss diagnostic imaging MeSH
- Incisor * abnormalities diagnostic imaging MeSH
- Age Factors MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Bone nonunion delays fracture end repair and is associated with inflammation. Although bone nonunion can be effectively repaired in clinical practice, many cases of failure. Studies have confirmed that BMP-2 and nHA/PA66 repaired bone defects successfully. There are few studies on the effects of the combined application of BMP-2 and NHA/PA66 on bone nonunion osteogenesis and inflammation. We aimed to investigate the expression level of inflammation-related genes in patients with bone nonunion and the effect of BMP-2-infected mesenchymal stem cells combined with nHA/PA66 on the level of inflammation in femur nonunion rats. We searched for a gene expression profile related to bone nonunion inflammation (GSE93138) in the GEO public database. Bone marrow mesenchymal stem cells (MSCs) of SD rats were cultured and passed through. We infected the third generation of MSCs with lentivirus carrying BMP-2 and induced the infected MSCs to bone orientation. We detected the expression level of BMP-2 by RT-PCR and the cell viability and alkaline phosphatase (ALP) activity by CCK8 and then analyzed the cell adhesion ability. Finally, the levels of related inflammatory factors, including C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and Erythrocyte Sedimentation Rate (ESR), were detected in nonunion rats. Our findings: The patients with nonunion had up-regulated expression of 26 differentially inflammatory genes. These genes are mainly enriched in innate immune response, extracellular region, calcium ion binding, Pantothenate and CoA biosynthesis pathways. The expression level of BMP-2 in the Lenti-BMP-2 group was higher (vs. empty lentivirus vector group: t=5.699; vs. uninfected group t=3.996). The cell activity of the MSCs + BMP-2 + nHA/PA66 group increased gradually. After being combined with nHA/PA66, MSCs transfected with BMP-2 spread all over the surface of nHA/PA66 and grew into the material pores. MSCs + BMP-2 + nHA/PA66 cells showed positive ALP staining, and the OD value of ALP was the highest. The levels of CRP, IL-6, TNF-alpha, and ESR in the MSCs + BMP-2 + nHA/PA66 group were lower than those in the MSCs and MSCs + nHA/PA66 group but higher than those in MSCs + BMP-2 group. The above comparisons were all P<0.05. The findings demonstrated that the expression level of inflammation-related genes increased in the patients with bone nonunion. The infection of MSCs by BMP-2 could promote the directed differentiation of MSCs into osteoblasts in the bone marrow of rats, enhance the cell adhesion ability and ALP activity, and reduce inflammation in rats with bone nonunion.
- MeSH
- Adult MeSH
- Femur metabolism pathology MeSH
- Femoral Fractures metabolism genetics MeSH
- Bone Morphogenetic Protein 2 * metabolism genetics MeSH
- Rats MeSH
- Cells, Cultured MeSH
- Middle Aged MeSH
- Humans MeSH
- Mesenchymal Stem Cells * metabolism MeSH
- Fractures, Ununited * genetics metabolism MeSH
- Osteogenesis MeSH
- Rats, Sprague-Dawley * MeSH
- Mesenchymal Stem Cell Transplantation * MeSH
- Inflammation * metabolism genetics MeSH
- Animals MeSH
- Check Tag
- Adult MeSH
- Rats MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
BACKGROUND: Acanthopanax senticosus (Rupr. et Maxim.) is commonly used in Traditional Chinese Medicine. Syringin is a major ingredient of phenolic glycoside in Acanthopanax senticosus. OBJECTIVE: This study was performed to investigate whether Syringin could protect high glucose-induced bone marrow mesenchymal stem cells (BMSCs) injury, cell senescence, and osteoporosis by inhibiting JAK2/STAT3 signaling. METHODS: BMSCs isolated from both the tibia and femur of mice were induced for osteogenesis. The cell senescence was induced using the high glucose medium. The cells were treated with 10 and 100 μmol/l Syringin. Immunohistochemistry staining was performed to determine the β-galactosidase (SA-β-gal) levels in differentially treated BMSCs. MTT assay and flow cytometry analysis were also performed to assess cell viability and cell cycle. The level of ROS in cells with different treatment was measured by using flow cytometry with DCF-DA staining. Calcium deposition and mineralized matrices were detected with alizarin red and ALP staining, respectively. Osteogenesis related genes OCN, ALP, Runx2, and BMP-2 were detected by RT-PCR. Levels of senescence-related proteins including p53 and p21, as well as JAK2, p-JAK2, STAT3, and p-STAT3 were detected by Western blot analysis. RESULTS: Syringin treatment reversed the phenotypes of senescence caused by high glucose in BMSCs, including the arrest of G0/G1 cell cycle, enhanced SA-β-gal activity, and impaired cell growth. Syringin also decreased the elevated ROS production and the levels of p53, p21, and JAK2/STAT3 signaling activation. In addition, Syringin also enhanced the osteogenic potential determined by ARS and ALP staining, as well as increasing OCN, ALP, Runx2, and BMP-2 expressions. CONCLUSION: Syringin protects high glucose-induced BMSC injury, cell senescence, and osteoporosis by inhibiting JAK2/STAT3 signaling.
- MeSH
- Phenylpropionates pharmacology MeSH
- Glucose * metabolism toxicity MeSH
- Glucosides * pharmacology MeSH
- Janus Kinase 2 * metabolism MeSH
- Mesenchymal Stem Cells * drug effects metabolism MeSH
- Mice MeSH
- Osteogenesis * drug effects MeSH
- Osteoporosis * prevention & control metabolism chemically induced pathology drug therapy MeSH
- Reactive Oxygen Species metabolism MeSH
- Signal Transduction * drug effects MeSH
- Cellular Senescence * drug effects MeSH
- STAT3 Transcription Factor * metabolism MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
INTRODUCTION: Fibroblast growth factor 20 (Fgf20), a member of the Fgf9 subfamily, was identified as an important regulator of bone differentiation and homeostasis processes. However, the role of Fgf20 in bone physiology has not been approached yet. Here we present a comprehensive bone phenotype analysis of mice with functional ablation of Fgf20. METHODS: The study conducts an extensive analysis of Fgf20 knockout mice compared to controls, incorporating microCT scanning, volumetric analysis, Fgf9 subfamily expression and stimulation experiment and histological evaluation. RESULTS: The bone phenotype could be detected especially in the area of the lumbar and caudal part of the spine and in fingers. Regarding the spine, Fgf20-/- mice exhibited adhesions of the transverse process of the sixth lumbar vertebra to the pelvis as well as malformations in the distal part of their tails. Preaxial polydactyly and polysyndactyly in varying degrees of severity were also detected. High resolution microCT analysis of distal femurs and the fourth lumbar vertebra showed significant differences in structure and mineralization in both cortical and trabecular bone. These findings were histologically validated and may be associated with the expression of Fgf20 in chondrocytes and their progenitors. Moreover, histological sections demonstrated increased bone tissue formation, disruption of Fgf20-/- femur cartilage, and cellular-level alterations, particularly in osteoclasts. We also observed molar dysmorphology, including root taurodontism, and described variations in mineralization and dentin thickness. DISCUSSION: Our analysis provides evidence that Fgf20, together with other members of the Fgf9 subfamily, plays a crucial regulatory role in skeletal development and bone homeostasis.
- MeSH
- Phenotype MeSH
- Fibroblast Growth Factors * metabolism genetics MeSH
- Calcification, Physiologic MeSH
- Bone and Bones metabolism pathology diagnostic imaging abnormalities MeSH
- Mice, Inbred C57BL MeSH
- Mice, Knockout * MeSH
- Mice MeSH
- Osteogenesis MeSH
- X-Ray Microtomography MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Patologická kalcifikace v kůži a v podkoží je poměrně heterogenním tématem. Z etiopatogenetického hlediska jsou patologické kalcifikace rozdělovány na dystrofické, metastatické, idiopatické a iatrogenní. Zvlášť je vyčleňována kalcifylaxe. Ektopická osifikace v dermatologii je jevem vzácným, vznikajícím buď jako izolovaný nález, či jako součást řady nádorů, pro které je osifikace typickým znakem. Práce shrnuje problematiku patologické kalcifikace a ektopické osifikace v dermatologii a dermatopatologii. Článek předkládá definici, stručný popis etiopatogeneze a výčet nejčastějších onemocnění spojených s jednotlivými typy patologického ukládání vápníku v kůži, podkoží a měkkých tkáních. Text dále shrnuje problematiku osteoma cutis z hlediska klinického obrazu a histopatologie. V závěru je uveden stručný přehled diagnostických a terapeutických možností.
Pathological calcification in the skin and subcutaneous tissue is relatively heterogenous issue. From the etiopathogenetic point of view, the pathological calcifications are divided into dystrophic, metastatic, idiopathic, and iatrogenic type. Calciphylaxis is distinguished as a distinctive type. Ectopic ossification in dermatology is a rare phenomenon, which arises as an isolated finding or as a part of the range of tumours, in which the ossification is a typical feature. The article summarizes the topic of the pathological calcification and the ectopic ossification in dermatology and dermatopathology. The paper presents the definition, the brief description of the etiopathogenesis and the list of the most common diseases connected to the particular types of the pathological calcium deposition in the skin, the subcutaneous tissue and the soft tissue. The review also summarizes the problematics of the osteoma cutis from the view of the clinical picture and the histopathology. Finally, the short summary of the diagnostic and therapeutic alternatives is discussed.
- Keywords
- osteoma cutis,
- MeSH
- Calcinosis * diagnosis etiology classification pathology therapy MeSH
- Skin pathology MeSH
- Humans MeSH
- Osteogenesis * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Review MeSH
- Examination Questions MeSH
Diamond-like carbon (DLC) layers are known for their high corrosion and wear resistance, low friction, and high biocompatibility. However, it is often necessary to dope DLC layers with additional chemical elements to strengthen their adhesion to the substrate. Ti-DLC layers (doped with 0.4, 2.1, 3.7, 6.6, and 12.8 at.% of Ti) were prepared by dual pulsed laser deposition, and pure DLC, glass, and polystyrene (PS) were used as controls. In vitro cell-material interactions were investigated with an emphasis on cell adhesion, proliferation, and osteogenic differentiation. We observed slightly increasing roughness and contact angle and decreasing surface free energy on Ti-DLC layers with increasing Ti content. Three-week biological experiments were performed using adipose tissue-derived stem cells (ADSCs) and bone marrow mesenchymal stem cells (bmMSCs) in vitro. The cell proliferation activity was similar or slightly higher on the Ti-doped materials than on glass and PS. Osteogenic cell differentiation on all materials was proved by collagen and osteocalcin production, ALP activity, and Ca deposition. The bmMSCs exhibited greater initial proliferation potential and an earlier onset of osteogenic differentiation than the ADSCs. The ADSCs showed a slightly higher formation of focal adhesions, higher metabolic activity, and Ca deposition with increasing Ti content.
