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International Journal of Molecular... Nov 2021We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs). (Comparative Study)
Comparative Study
Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method.
BACKGROUND
We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs).
METHOD
We isolated DFAT cells and ASCs from GFP mice. DFAT cells were established by a new culture method using a mesh culture instead of a ceiling culture. The isolated DFAT cells and ASCs were incubated in osteogenic medium, then alizarin red staining, alkaline phosphatase (ALP) assays, and RT-PCR (for RUNX2, osteopontin, DLX5, osterix, and osteocalcin) were performed to evaluate the osteoblastic differentiation ability of both cell types in vitro. In vivo, the DFAT cells and ASCs were incubated in osteogenic medium for four weeks and seeded on collagen composite scaffolds, then implanted subcutaneously into the backs of mice. We then performed hematoxylin and eosin staining and immunostaining for GFP and osteocalcin.
RESULTS
The alizarin red-stained areas in DFAT cells showed weak calcification ability at two weeks, but high calcification ability at three weeks, similar to ASCs. The ALP levels of ASCs increased earlier than in DFAT cells and showed a significant difference ( < 0.05) at 6 and 9 days. The ALP levels of DFATs were higher than those of ASCs after 12 days. The expression levels of osteoblast marker genes (osterix and osteocalcin) of DFAT cells and ASCs were higher after osteogenic differentiation culture.
CONCLUSION
DFAT cells are easily isolated from a small amount of adipose tissue and are readily expanded with high purity; thus, DFAT cells are applicable to many tissue-engineering strategies and cell-based therapies.
Topics: Adipocytes; Adipose Tissue; Animals; Bone Regeneration; Calcification, Physiologic; Cell Culture Techniques; Cell Dedifferentiation; Cell Differentiation; Cells, Cultured; Green Fluorescent Proteins; Mice; Mice, Transgenic; Osteoblasts; Osteogenesis; Stem Cell Transplantation; Stem Cells; Tissue Engineering; Transplantation, Autologous
PubMed: 34830277
DOI: 10.3390/ijms222212392 -
Journal of Clinical Hypertension... Jul 2022Osteocalcin (OCN) is a bone-derived and vitamin K dependent hormone that affects energy metabolism and vascular calcification. The relationship between serum OCN and...
Osteocalcin (OCN) is a bone-derived and vitamin K dependent hormone that affects energy metabolism and vascular calcification. The relationship between serum OCN and vascular function in patients with chronic kidney disease (CKD) is uncertain. This study investigated the association between serum OCN and vascular function as expressed with reactive hyperemia index (RHI) and augmentation index (AIx) measured by Endo-PAT 2000 device. This cross-sectional analysis was based on 256 pre-dialysis CKD patients who had completed the Endo-PAT 2000 test and serum OCN at the First Center of Chinese PLA Hospital from November 2017 to December 2019. Based on whether the RHI was less than 1.67, the patients were divided into endothelial dysfunction and normal endothelial function groups. Multiple logistic and linear regression were used to analyze the association between OCN and vascular function. Subgroup analyses were performed to examine the effects of OCN on vascular function in different CKD populations. After multivariate adjustment, CKD with low OCN were more likely to have endothelial dysfunction (OR: 0.794; 95%CI: 0.674-0.934; P = .006); on the contrary, patients with high OCN had a higher degree of arterial stiffness (standardized β: 0.174; P = .003). Subgroup analyses showed that higher OCN was associated with severe arterial stiffness but a better endothelial function in young (age < 65 years, P /P = .027/.011), male (P /P = .040/.016), patients with a history of hypertension (P /P = .004/.009) or diabetes (P /P = .005/.005), and in early CKD (P /P = .014/.015). In conclusion, serum OCN correlates with vascular function in CKD patients: beneficial for endothelial function but detrimental to arterial stiffness.
Topics: Aged; Cross-Sectional Studies; Endothelium, Vascular; Humans; Hyperemia; Hypertension; Male; Osteocalcin; Renal Insufficiency, Chronic; Vascular Stiffness
PubMed: 35687487
DOI: 10.1111/jch.14523 -
BMC Musculoskeletal Disorders Aug 2019Treatment of anterior cruciate ligament injuries commonly involves the use of polyethylene terephthalate (PET) artificial ligaments for reconstruction. However, the...
BACKGROUND
Treatment of anterior cruciate ligament injuries commonly involves the use of polyethylene terephthalate (PET) artificial ligaments for reconstruction. However, the currently available methods require long fixation periods, thereby necessitating the development of alternative methods to accelerate the healing process between tendons and bones. Thus, we developed and evaluated a novel technique that utilizes silicate-substituted strontium (SrSiP).
METHODS
PET films, nano-coated with SrSiP, were prepared. Bone marrow mesenchymal cells (BMSCs) from femurs of male rats were cultured and seeded at a density of 1.0 × 10/cm onto the SrSiP-coated and non-coated PET film, and subsequently placed in an osteogenic medium. The osteocalcin concentration secreted into the medium was compared in each case. Next, PET artificial ligament, nano-coated with SrSiP, were prepared. BMSCs were seeded at a density of 4.5 × 10/cm onto the SrSiP-coated, and non-coated artificial ligament, and then placed in osteogenic medium. The osteocalcin and calcium concentrations in the culture medium were measured on the 8th, 10th, 12th, and 14th day of culture. Furthermore, mRNA expression of osteocalcin, alkaline phosphatase (ALP), bone morphogenetic protein-2 (BMP2), and runt-related transcription factor 2 (Runx2) was evaluated by qPCR. We transplanted the SrSiP-coated and non-coated artificial ligament to the tibiae of mature New Zealand white rabbits. Two months later, we sacrificed them and histologically evaluated them.
RESULTS
The secretory osteocalcin concentration in the medium on the film was significantly higher for the SrSiP group than for the non-coated group. Secretory osteocalcin concentration in the medium on the artificial ligament was also significantly higher in the SrSiP group than in the non-coated group on the 14th day. Calcium concentration on the artificial ligament was significantly lower in the SrSiP group than in the non-coated group on the 8th, 10th, 12th, and 14th day. In qPCR as well, OC, ALP, BMP2, and Runx2 mRNA expression were significantly higher in the SrSiP group than in the non-coated group. Newly formed bone was histologically found around the artificial ligament in the SrSiP group.
CONCLUSIONS
Our findings demonstrate that artificial ligaments using SrSiP display high osteogenic potential and thus may be efficiently used in future clinical applications.
Topics: Animals; Anterior Cruciate Ligament Injuries; Apatites; Bone-Implant Interface; Calcium; Cell Differentiation; Cells, Cultured; Coated Materials, Biocompatible; Culture Media; Disease Models, Animal; Humans; Male; Materials Testing; Mesenchymal Stem Cells; Nanostructures; Osseointegration; Osteocalcin; Osteogenesis; Polyethylene Terephthalates; Primary Cell Culture; Rabbits; Rats; Silicates; Strontium; Time Factors; Wound Healing
PubMed: 31472679
DOI: 10.1186/s12891-019-2777-8 -
European Review For Medical and... Nov 2022Our aim is to investigate the correlation between risk factors of postmenopausal osteoporotic fracture, BMD and Bone turnover markers, lipid metabolism and BMI.
Correlation analysis between risk factors, BMD and serum osteocalcin, CatheK, PINP, β-crosslaps, TRAP, lipid metabolism and BMI in 128 patients with postmenopausal osteoporotic fractures.
OBJECTIVE
Our aim is to investigate the correlation between risk factors of postmenopausal osteoporotic fracture, BMD and Bone turnover markers, lipid metabolism and BMI.
SUBJECTS AND METHODS
The Cox proportional hazard model was used to conduct univariate and multivariate analysis to screen the risk factors related to postmenopausal osteoporotic fractures. Blood samples were collected to detect biochemical markers of bone turnover, blood lipids content, and then measure the BMI of the survey subjects. BMD was measured and its correlation with biochemical markers of bone turnover, lipid metabolism and BMI was analyzed.
RESULTS
Cox univariate analysis indicated that average age, menopause, years since menopause, number of deliveries, and limb spasm are associated covariates of postmenopausal osteoporotic fractures. Where, BMD severity, history of hysterectomy or ovariectomy, and years since menopause are significant covariates for the incidence of postmenopausal osteoporotic fractures. The correlation study with lipid metabolism found that the smaller the BMI value, the greater the BMD loss; the smaller the TG value, the greater the BMD loss, exhibiting a downward trend. No difference was observed between HDL-C and LDL-C content, and the difference was not statistically significant (p>0.05). Femoral neck BMD was negatively correlated with CatheK, serum osteocalcin, PINP, β-crosslaps and TRAP, and lumbar spine BMD was also negatively correlated with CatheK, serum osteocalcin, PINP, β-crosslaps and TRAP.
CONCLUSIONS
Biochemical markers of bone turnover are highly expressed in postmenopausal women and increase with the decrease of bone density, which can be used as markers for disease prediction. Combined with BMI, triglyceride and other related indicators, and closely related factors such as the patient's age, the number of deliveries, it is possible to predict the incidence of PMOP fractures early.
Topics: Female; Humans; Biomarkers; Body Mass Index; Correlation of Data; Lipid Metabolism; Osteocalcin; Osteoporosis, Postmenopausal; Osteoporotic Fractures; Postmenopause; Risk Factors; Dental Enamel Proteins
PubMed: 36394744
DOI: 10.26355/eurrev_202211_30147 -
International Journal of Molecular... Feb 2023It is well known that patients with attention deficit hyperactivity disorder treated with stimulants, such as methylphenidate hydrochloride (MPH), have reduced height...
It is well known that patients with attention deficit hyperactivity disorder treated with stimulants, such as methylphenidate hydrochloride (MPH), have reduced height and weight. Even though MPH has an anorexigenic effect, an additional impact of this drug on the growth plate cannot be discarded. In this study, we aimed to determine the cellular effect of MPH on an in vitro growth plate model. We tested the effects of MPH on the viability and proliferation of a prechondrogenic cell line via an MTT assay. In vitro differentiation of this cell line was performed, and cell differentiation was evaluated through the expression of cartilage- and bone-related genes as measured via RT-PCR. MPH did not alter the viability or proliferation of prechondrogenic cells. However, it reduced the expression of cartilage extracellular matrix-related genes (type II collagen and aggrecan) and increased the expression of genes involved in growth plate calcification (Runx2, type I collagen, and osteocalcin) at different phases of their differentiation process. Our results evidence that MPH upregulates genes associated with growth plate hypertrophic differentiation. This may induce premature closure of the growth plate, which would contribute to the growth retardation that has been described to be induced by this drug.
Topics: Humans; Attention Deficit Disorder with Hyperactivity; Central Nervous System Stimulants; Growth Plate; Methylphenidate; Osteogenesis; Cells, Cultured
PubMed: 36835608
DOI: 10.3390/ijms24044175 -
Nutrients Jan 2022Osteoporosis is a major health concern in aging populations, where 54% of the U.S. population aged 50 and older have low bone mineral density (BMD). Increases in... (Randomized Controlled Trial)
Randomized Controlled Trial
Osteoporosis is a major health concern in aging populations, where 54% of the U.S. population aged 50 and older have low bone mineral density (BMD). Increases in inflammation and oxidative stress play a major role in the development of osteoporosis. Men are at a greater risk of mortality due to osteoporosis-related fractures. Our earlier findings in rodent male and female models of osteoporosis, as well as postmenopausal women strongly suggest the efficacy of prunes (dried plum) in reducing inflammation and preventing/reversing bone loss. The objective of this study was to examine the effects of two doses of prunes, daily, on biomarkers of inflammation and bone metabolism in men with some degree of bone loss (BMD; t-score between -0.1 and -2.5 SD), for three months. Thirty-five men between the ages of 55 and 80 years were randomized into one of three groups: 100 g prunes, 50 g prunes, or control. Consumption of 100 g prunes led to a significant decrease in serum osteocalcin ( < 0.001). Consumption of 50 g prunes led to significant decreases in serum osteoprotegerin (OPG) ( = 0.003) and serum osteocalcin ( = 0.040), and an increase in the OPG:RANKL ratio ( = 0.041). Regular consumption of either 100 g or 50 g prunes for three months may positively affect bone turnover.
Topics: Aged; Aged, 80 and over; Biomarkers; Body Composition; Bone Density; Bone Remodeling; Bone and Bones; Exercise; Humans; Inflammation; Lumbar Vertebrae; Male; Middle Aged; Osteocalcin; Osteoporosis; Osteoporotic Fractures; Osteoprotegerin; Phytotherapy; Prunus domestica; RANK Ligand
PubMed: 35057457
DOI: 10.3390/nu14020276 -
Brazilian Oral Research 2023This study aimed to investigate whether GSK-3 inhibition (CHIR99021) effectively promoted mineralization by cementoblasts (OCCM-30). OCCM-30 cells were used and treated...
This study aimed to investigate whether GSK-3 inhibition (CHIR99021) effectively promoted mineralization by cementoblasts (OCCM-30). OCCM-30 cells were used and treated with different concentrations of CHIR99021 (2.5, 5, and 10 mM). Experiments included proliferation and viability, cellular metabolic activity, gene expression, and mineral nodule formation by Xylene Orange at the experimental time points. In general, CHIR99021 did not significantly affect OCCM-30 viability and cell metabolism (MTT assay) (p > 0.05), but increased OCCM-30 proliferation at 2.5 mM on days 2 and 4 (p < 0.05). Data analysis further showed that inhibition of GSK-3 resulted in increased transcript levels of Axin2 in OCCM-30 cells starting as early as 4 h, and regulated the expression of key bone markers including alkaline phosphatase (Alp), runt-related transcription factor 2 (Runx-2), osteocalcin (Ocn), and osterix (Osx). In addition, CHIR99021 led to an enhanced mineral nodule formation in vitro under both osteogenic and non-osteogenic conditions as early as 5 days after treatment. Altogether, the results of the current study suggest that inhibition of GSK-3 has the potential to promote cementoblast differentiation leading to increased mineral deposition in vitro.
Topics: Dental Cementum; Glycogen Synthase Kinase 3; Cell Proliferation; Osteocalcin; Cell Differentiation
PubMed: 37970932
DOI: 10.1590/1807-3107bor-2023.vol37.0112 -
European Review For Medical and... Aug 2022We explored the influences on platelet-rich fibrin (PRF) to rat Bone Mesenchymal Stem Cells (BMSCs), as well as the role of bone morphogenetic protein 2 (BMP2)/maternal...
OBJECTIVE
We explored the influences on platelet-rich fibrin (PRF) to rat Bone Mesenchymal Stem Cells (BMSCs), as well as the role of bone morphogenetic protein 2 (BMP2)/maternal signal protein homolog (Smads) pathway.
MATERIALS AND METHODS
The proposed research is approved by the ethics board of the Second Affiliated Hospital of Harbin Medical University. The BMSCs were isolated and purified. The BMSCs were assigned to a control group arbitrarily, PRF group, BMP activator group and BMP inhibitor group (hereinafter referred to as activator group and inhibitor group). Each group of BMSCs in the logarithmic growth phase was detected for the alkaline phosphatase (ALP) activity since 3 days and 14 days of culture; CCK-8 assay was conducted for detection of the proliferation of BMSCs; Real time PCR was conducted for detection of the osteogenic differentiation marker collagen I (COL-I), BMP2, Runt-related transcription factor 2(RUNX2), osteocalcin (OCN) mRNA relative expression levels; Western-Blot detection of BMP2, OCN, P-SMAD1/5/8, relative expression level of RUNX2 protein.
RESULTS
In contrast to the control group, BMSCs' the ALP activity of the PRF group, activator group, as well as inhibitor group increased for 3 days and 14 days, and the activator group>PRF group>inhibitor group (p≤0.05). ALP activity in each group was elevated with the increase in culture time, the ALP activity of the control group, PRF group, activator group and inhibitor group increased (p≤0.05). In comparison to the control group, the relevant expression levels of COL-I, BMP-2, RUNX2 and OCN in the PRF group, activator group, and inhibitor group increased, and the activator group>PRF group>inhibitor group (p≤0.05). The relative expression levels of BMP2, OCN, p-SMAD1/5/8 and RUNX2 protein in each group were statistically different, the activator group>PRF group>control group>inhibitor group (p≤0.05).
CONCLUSIONS
PRF can promote the proliferation and osteogenic differentiation of BMSCs by activating the BMP2/Smads signaling pathway.
Topics: Animals; Bone Morphogenetic Protein 2; Collagen Type I; Core Binding Factor Alpha 1 Subunit; Humans; Mesenchymal Stem Cells; Osteocalcin; Osteogenesis; Platelet-Rich Fibrin; Rats
PubMed: 35993636
DOI: 10.26355/eurrev_202208_29409 -
Journal of Dental Sciences Jul 2022Tristrontium aluminate (SA) is a hydraulic cement with setting behavior similar to that of mineral trioxide aggregate (MTA). This study examined the biological effects...
BACKGROUND/PURPOSE
Tristrontium aluminate (SA) is a hydraulic cement with setting behavior similar to that of mineral trioxide aggregate (MTA). This study examined the biological effects of SA on mouse dental papilla cells (MDPs) and on rat exposed pulps .
MATERIALS AND METHODS
Extracts of SA and MTA were prepared by immersing each cement in ultrapure water. MDPs were cultured with SA or MTA extracts, and cell proliferation was evaluated with a tetrazolium-salt assay. Attachment of MDPs on the set cements was examined with scanning electron microscopy (SEM). mRNA expression of bone morphogenic protein (Bmp2), osteocalcin (Oc) and osteopontin (Opn) in MDPs exposed to SA or MTA extracts was determined with reverse transcription-quantitative polymerase chain reaction. Mineralized nodule formation was evaluated with Alizarin Red S staining. Simulated body fluid (SBF)-dipped SA was examined with SEM and energy dispersive X-ray analysis (EDX). Exposed molar pulps of male Wistar rats capped with SA or MTA were histologically examined.
RESULTS
SA extract did not inhibit proliferation of MDPs. Set SA and MTA exhibited attachment of MDPs on their surface. SA extract showed significantly higher mineralized nodule formation and mRNA expression of Bmp2, Oc, and Opn than did MTA extract. SBF-dipped SA exhibited formation of surface precipitates, which were composed of Ca, P, Sr, and Al. Direct pulp capping with SA and with MTA induced mineralized tissue repair of the exposed pulp.
CONCLUSION
SA possesses biocompatibility and pro-mineralization effects comparable to those of MTA.
PubMed: 35784112
DOI: 10.1016/j.jds.2021.12.018 -
Bioengineered Dec 2021MicroRNAs (miRNAs) regulate osteogenic differentiation and influence osteoporosis (OP). The aim of this study was to determine the potential role of miR-874-3p in OP....
MicroRNAs (miRNAs) regulate osteogenic differentiation and influence osteoporosis (OP). The aim of this study was to determine the potential role of miR-874-3p in OP. The expression levels of miR-874-3p and leptin (LEP) in the femoral neck trabeculae of 35 patients with or without OP were measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The effects of miR-874-3p or LEP on the cell proliferation and alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osterix (OSX) levels were observed by upregulating miR-874-3p in human bone marrow mesenchymal stem cells (hBMSCs). Additionally, calcium deposition levels were evaluated using alizarin red staining (ARS). Molecular mechanisms of miR-874-3p and LEP underlying the osteogenic differentiation of hBMSCs were also evaluated using bioinformatics analysis, luciferase reporter assays, and RNA pull-down assays. The miR-874-3p levels were significantly lower in the femoral neck trabeculae of patients with OP than those of the control group, while the opposite was observed regarding the levels of LEP. Expression levels of miR-874-3p in hBMSCs were upregulated during osteogenic differentiation, while those of LEP were downregulated. Moreover, miR-874-3p upregulation promoted ALP, RUNX2, OCN, and OSX mRNA expression, cell proliferation, and calcium deposition in hBMSCs. LEP was found to be a target gene of miR-874-3p. Overexpression of LEP inhibited the expression of osteoblast markers and reversed the effect of osteogenic differentiation induced by the upregulation of miR-874-3p. In conclusion, miR-874-3p promoted the proliferation and differentiation of hBMSCs by downregulating the expression of LEP, thus inhibiting OP. miRNAs: microRNAs; OP: osteoporosis; hBMSCs: human Bone Marrow Mesenchymal stem cells; LEP: leptin; DEGs: differentially expressed genes.
Topics: Cell Differentiation; Gene Expression Regulation, Neoplastic; Gene Ontology; Humans; Leptin; Mesenchymal Stem Cells; MicroRNAs; Osteoblasts; Osteogenesis; Osteoporosis
PubMed: 34818977
DOI: 10.1080/21655979.2021.2009618