Prevention of methylprednisolone acetate-induced osteoporosis with calcium administration in rat model

Glucocorticoid steroids are widely used as anti-inflammatory and immunosuppressive medications and are well known to induce osteoporosis. In Present study 24 rats were randomly divided into four groups (n=6): Group A (control), Group B (sham)that was treated only by normal saline for 1 month.Group C that was treated by methylprednisolone acetate alone (0.2 mg/kg) for 1 month. Group D that was treated by methylprednisolone acetate (0.2 mg/kg) and oral calcium supplementation (15 mg/kg) for 1 month. Changes in concentration of bone metabolic markers such as osteocalcine, acid phosphatase and calcium were evaluated before and after treatment. Bone mineral density (BMD) of lumbar vertebrae was also measured by dual energy X ray absorptiometry (DEXA). The results showed that concentration mean of serum acid phosphatase was increased significantly (P � 0.05) in C and D groups in compared to A and B groups. The concentration mean of serum osteocalcine in group C was decreased significantly (P � 0.05) in comparison to A and B groups but increased significantly in the group D in comparison to group C. The concentration mean of serum calcium was decreased significantly (P � 0.05) in C and D groups in compared to A and B groups. The bone mineral density (g/cm2) was decreased significantly (P � 0.05) in group C in compared to A and B groups. This increased significantly in group D in compared to group C. These results are compatible with the view that low doses of methylprednisolone acetate decreases bone formation and increase bone resorption in the lumbar vertebrae of rats. Calcium administration decreased effects of methylprednisolone. © 2009 Tehran University of Medical Sciences. All rights reserved

Melatonin pretreated blastocysts along with calcitonin administration improved implantation by upregulation of heparin binding-epidermal growth factor expression in murine endometrium

Objective: Implantation failure is an obstacle in assisted reproduction techniques (ART). Calcitonin is a molecules involved in uterine receptivity and embryo implantation. Melatonin can promote embryo quality and improve implantation. This study examines the effect of pretreatment of blastocysts with melatonin and calcitonin on heparin binding-epidermal growth factor (HB-EGF) expression in murine endometrium. Materials and Methods: In this experimental study, we collected 2-cell embryos from the oviducts of 1.5 day pregnant NMRI mice. Embryos were cultured to the blastocyst in GTM medium with or without 10-9 M melatonin. Pregnant and pseudo-pregnant mice received intraperitoneal (IP) injections of 2 IU calcitonin. After 24 hours, we transferred the cultured blastocysts into the uteri of pseudo-pregnant mice. Two days later, implantation sites were counted and we assessed the levels of HB-EGF mRNA and protein in the uteri of naturally pregnant and pseudo-pregnant mice by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Statistical analysis was performed with one-way ANOVA followed by the Tukey post hoc test. P<0.05 was considered statistically significant. Results: Melatonin pretreatment of blastocysts along with calcitonin administration significantly increased HB-EGF mRNA and protein (P<0.001) in the endometrium of pseudo-pregnant mice. Administration of calcitonin in naturally pregnant mice significantly increased HB-EGF mRNA and protein levels (P<0.001). Compared with the control group (2.6 ± 0.5), the average number of implantation sites in the melatonin group (4.6 ± 0.5, P<0.05) and calcitonin group (7 ± 1, P<0.001) significantly increased. There was a significant increase in implantation sites in the combined melatonin and calcitonin group (8.6 ± 0.5, P<0.001). Calcitonin significantly enhanced calcitonin receptor mRNA (P<0.001) and protein (P<0.05) in the uteri of naturally pregnant and pseudo-pregnant mice. Conclusion: Melatonin pretreated blastocysts along with calcitonin increased HB-EGF expression in the uteri of pseudopregnant mice. Calcitonin administration upregulated HB-EGF in uteri of naturally pregnant mice

Melatonin pretreated blastocysts along with calcitonin administration improved implantation by upregulation of heparin binding-epidermal growth factor expression in murine endometrium

Objective: Implantation failure is an obstacle in assisted reproduction techniques (ART). Calcitonin is a molecules involved in uterine receptivity and embryo implantation. Melatonin can promote embryo quality and improve implantation. This study examines the effect of pretreatment of blastocysts with melatonin and calcitonin on heparin binding-epidermal growth factor (HB-EGF) expression in murine endometrium. Materials and Methods: In this experimental study, we collected 2-cell embryos from the oviducts of 1.5 day pregnant NMRI mice. Embryos were cultured to the blastocyst in GTM medium with or without 10-9 M melatonin. Pregnant and pseudo-pregnant mice received intraperitoneal (IP) injections of 2 IU calcitonin. After 24 hours, we transferred the cultured blastocysts into the uteri of pseudo-pregnant mice. Two days later, implantation sites were counted and we assessed the levels of HB-EGF mRNA and protein in the uteri of naturally pregnant and pseudo-pregnant mice by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Statistical analysis was performed with one-way ANOVA followed by the Tukey post hoc test. P&lt;0.05 was considered statistically significant. Results: Melatonin pretreatment of blastocysts along with calcitonin administration significantly increased HB-EGF mRNA and protein (P&lt;0.001) in the endometrium of pseudo-pregnant mice. Administration of calcitonin in naturally pregnant mice significantly increased HB-EGF mRNA and protein levels (P&lt;0.001). Compared with the control group (2.6 ± 0.5), the average number of implantation sites in the melatonin group (4.6 ± 0.5, P&lt;0.05) and calcitonin group (7 ± 1, P&lt;0.001) significantly increased. There was a significant increase in implantation sites in the combined melatonin and calcitonin group (8.6 ± 0.5, P&lt;0.001). Calcitonin significantly enhanced calcitonin receptor mRNA (P&lt;0.001) and protein (P&lt;0.05) in the uteri of naturally pregnant and pseudo-pregnant mice. Conclusion: Melatonin pretreated blastocysts along with calcitonin increased HB-EGF expression in the uteri of pseudopregnant mice. Calcitonin administration upregulated HB-EGF in uteri of naturally pregnant mice

PROTECTIVE EFFECT OF VITAMIN D3 IN METHYLPREDNISOLONE ACETATE (MPA) INDUCED LOSS OF BONE METABOLISM MARKERS AND BONE MINERAL DENSITY IN THE LUMBAR SPINE OF RAT

Although some vitamins have been shown to prevent glucocorticoids induced osteoporosis in short time, the magnitude of this effect remains to be clarified. The aim of this study was to evaluate protective effect of vitamin D3 on methylprednisolone acetate (MPA) induced osteoporosis in rats. Twenty-four male Sprague Dawley rats were randomly divided into four groups: Group A (n = 6), was a base line control or normal animals. Group B (n = 6), was treated only normal saline, group C (n = 6), was treated MPA (0.2 mg/kg) subcutaneously for 4 weeks (3 times per a week) and finally group D (n = 6) were administered MPA resemble to group C and treated by Vitamin D3 (0.1 µg/kg dissolved in ethanol daily). Level of calcium, osteocalcin and acid phosphatase in serum were measured before and after treatment. Also, bone mineral density (BMD) of lumber vertebrae was measured by dual energy X-ray absorptiometry. The results showed that the serum calcium level unaffected by MPA in all groups before and after treatment, but the serum osteocalcin level and bone mineral density of lumbar vertebrae were significantly (P &lt; 0.05) decreased in group C compared with groups A and B. In group D serum osteocalcin level increased again significantly (P &lt; 0.05) but increasing of BMD and bone mineral content were not significant. The findings indicate that by using of vitamin D3 in MPA treated rats could increase bone formation and decrease bone resorption

The effects of progestrone on the in-vitro expression of P0, S100 and Krox20 genes in adipose-derived stem cells

&quot;n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Adipose-derived stem cells (ADSCs) have noticeable self-renewal ability and can differentiate into several cell lines such as adipocytes, osteoblasts, chondrocytes, and myocytes. Progesterone plays a significant role in the myelination of peripheral nerves. Regarding the role of progesterone on the myelination of peripheral nervous system, we evaluated its effects on the in-vitro expression of P0, S100 and Krox20 mRNA in adipose-derived stem cells.&quot;n&quot;nMethods : In this experimental study, rat adipose-derived stem cells were isolated from the inguinal region of the animals and were evaluated by flow cytometry before culture. In preinduction phase, the cells were sequentially treated with various factors such as &amp;beta;-mercaptoethanol and all-trans-retinoic acid, followed by different induction mixtures. &amp;nbsp;The cells were divided into four groups including two control groups (receiving either fibroblast and platelet derived-growth factors, or fibroblast growth factor, platelet derived-growth factor, forskolin and heregulin) and two experimental groups (receiving either fibroblast growth factor, platelet derived-growth factor, forskolin and progesterone, or fibroblast growth factor, platelet derived-growth factor, heregulin and progesterone). Expression of Schwann cell markers, S-100, P0 and Krox20 mRNA, was determined by semi-quantitative RT-PCR.&quot;n&quot;nResults : ADSCs expressed CD90, CD73, and CD31 but showed lack of CD45, and VEGFR2 expression. After the induction stage, S-100, P0 and Krox20 mRNA were expressed in the progesterone receiving experimental groups, but expression of S-100 and Krox20 mRNA were less than the control group which was receiving forskolin and heregulin (P&amp;lt;0.0001). &quot;n&quot;nConclusion: Progesterone can promote the in-vitro expression of S-100, P0, and Krox20 genes in adipose-derived stem cells

Effect of laminated hydroxyapatite/gelatin nanocomposite scaffold structure on osteogenesis using unrestricted somatic stem cells in rat

Bone matrix consists of two major phases at the nanoscale: organic and hydroxyapatite. Nanotechnology as a diverse and interdisciplinary area of research has the capacity to revolutionise many areas of applications such as bone tissue engineering. Nanohydroxyapatite/gelatin composite has higher osteoblast attachment and proliferation than micro-sized ones, and shorter culturing period and lower cell seeding density compared to pure gelatin. A nanostructured scaffold was fabricated by three methods for bone repair using nanohydroxyapatite and gelatin as the main components. Its biocompatibility, alizarin red test on the 14th and 21st days, gene expression on the 21st day in in vitro using and histomorphometry after 4 and 8 weeks postimplantation in the rat were investigated. Cultured unrestricted somatic stem cells used for in vitro study showed an excellent level of cell attachment to the scaffold. Cells induced more osteoblast differentiation on the scaffold than in 2D cell culture. Osteoblast differentiation and bone regeneration results of in vitro and in vivo investigation on scaffold were extremely significant, better than control and treatment groups. These effects could be attributed to the shape and size of nanoHA particles and good architecture of the scaffold. The results confirm the feasibility of bone regeneration using synthesised scaffold as a temporary bone substitute. © 2013 International Federation for Cell Biology

The effects of progestrone on the in-vitro expression of P0, S100 and Krox20 genes in Adipose-derived stem cells

Background: Adipose-derived stem cells (ADSCs) have noticeable self-renewal ability and can differentiate into several cell lines such as adipocytes, osteoblasts, chondrocytes, and myocytes. Progesterone plays a significant role in the myelination of peripheral nerves. Regarding the role of progesterone on the myelination of peripheral nervous system, we evaluated its effects on the in-vitro expression of P0, S100 and Krox20 mRNA in adipose-derived stem cells. Methods: In this experimental study, rat adipose-derived stem cells were isolated from the inguinal region of the animals and were evaluated by flow cytometry before culture. In preinduction phase, the cells were sequentially treated with various factors such as β- mercaptoethanol and all-trans-retinoic acid, followed by different induction mixtures. The cells were divided into four groups including two control groups (receiving either fibroblast and platelet derived-growth factors, or fibroblast growth factor, platelet derived-growth factor, forskolin and heregulin) and two experimental groups (receiving either fibroblast growth factor, platelet derived-growth factor, forskolin and progesterone, or fibroblast growth factor, platelet derived-growth factor, heregulin and progesterone). Expression of Schwann cell markers, S-100, P0 and Krox20 mRNA, was determined by semi-quantitative RT-PCR. Results: ADSCs expressed CD90, CD73, and CD31 but showed lack of CD45, and VEGFR2 expression. After the induction stage, S-100, P0 and Krox20 mRNA were expressed in the progesterone receiving experimental groups, but expression of S-100 and Krox20 mRNA were less than the control group which was receiving forskolin and heregulin (P<0.0001). Conclusion: Progesterone can promote the in-vitro expression of S-100, P0, and Krox20 genes in adipose-derived stem cells

Evaluation of the effect of adipose tissue-derived stem cells on the quality of bone healing around implants

Purpose/Aim: This study evaluates the efficacy of grafted adipose-derived stem cells (ADSCs) on blade-type implants in improving osseointegration in rat femurs using a low-densitybone model. Materials and Methods: After isolating and expanding ADSCs, twice-passaged cells were seeded on blade-type implants on culture plates. Osteogenic induction of grafted cells began after attaching cells to the prepared titanium surfaces and it continued for 4 days. The scaffolds were then implanted in the femurs of Wistar rats. Osteogenic differentiation of these cells was confirmed using polymerase chain reaction (PCR) and alizarin red staining ofthe mineralized extracellular matrix. After 8 weeks, histological and histomorphometric evaluations of undecalcified resin sections (bone-implant contact BIC % and bone mineral index BMI) were performed using light microscopy and scanning electron microscopy. Results: Alizarin red staining in conjunction with gene expression results confirmed osteogenic differentiation. Histomorphometric assessment using scanning electron microscopy demonstrated improved BIC% and BMI near the treated surface compared with the untreated surface. Conclusions: The complex of differentiated grafted ADSCs and extracellular matrix and the macrodesign and microdesign of the implant can improve osseointegration in low-density bone. © 2016 Taylor & Francis

Mesenchymal stem cell mediated effects on microglial phenotype in cuprizone-induced demyelination model

Microglial cells have an essential role in neurodegenerative disorders, such as multiple sclerosis. They are divided into two subgroups: M1 and M2 phenotypes. Mesenchymal stem cells (MSC), with neuroprotective and immunomodulating properties, could improve these diseases. We evaluate the immunomodulating effects of MSC on microglial phenotypes and the improvement of demyelination in a cuprizone (CPZ) model of multiple sclerosis (MS). For inducing the chronic demyelination model, C57BL6 mice were given a diet with 0.2 CPZ (w/w) for 12 weeks. In the MSC group, cells were transplanted into the right lateral ventricle of mice. The expression of targeted genes was assessed by real-time polymerase chain reaction. M1 and M2 microglial phenotypes were assessed by immunohistochemistry of inducible nitric oxide synthase (iNOS) and Arg-1, respectively. Remyelination was studied by luxal fast blue (LFB) staining and electron microscopy (EM). We found that MSC transplantation reduced the expression level of M1-specific messenger RNA (mRNA; iNOS and CD86) but increased the expression level of M2 specific genes (CD206, Arg-1, and CX3CR1) in comparison to the CPZ group. Moreover, cell therapy significantly decreased the M1 marker (iNOS+ cells), but M2 marker (Arg-1+ cells) significantly increased in comparison with the CPZ group. In addition, MSC treatment significantly increased the CX3CL1 expression level in comparison with the CPZ group and led to improvement in remyelination, which was confirmed by LFB and EM images. The results showed that MSC transplantation increases the M2 and decreases the M1 phenotype in MS. This change was accompanied by decrease in demyelination and axonal injury and indicated that MSCs have a positive effect on MS by modification of microglia cells. © 2019 Wiley Periodicals, Inc