Adipose derived stem cells affect miR-145 and p53 expressions of co-cultured hematopoietic stem cells

Objective: umbilical cord blood is used for transplantation purposes in regenerative medicine of hematological disorders. MicroRNAs are important regulators of gene expression that control both physiological and pathological processes such as cancer development and incidence. There is a new relation between p53 [tumor suppressor gene] and miR-145 [suppressor of cell growth] upregulation. In this study, we have assessed how adipose-derived stem cells [ADSCs] affect the expansion of hematopoietic stem cells [HSCs], as well as miR-145 and p53 expressions

Materials and Methods: in this experimental study, we cultured passage-3 isolated human ADSCs as a feeder layer. Flow cytometry analysis confirmed the presence of ADSC surface markers CD73, CD90, CD105. Ex vivo cultures of cordblood CD34+ cells were cultured under the following 4 culture conditions for 7 days: i. Medium only supplemented with cytokines, ii. Culture on an ADSCs feeder layer, iii. Indirect culture on an ADSCs feeder layer [Thin Cert[™] plate with a 0.4 ?m pore size], and iv. Control group analyzed immediately after extraction. Real-time polymerase chain reaction [PCR] was used to determine the expressions of the p53 and miR-145 genes. Flow cytometry analysis of cells stained by annexin V and propidium iodide [PI] was performed to detect the rate of apoptosis in the expanded cells

Results: ADSCs tested positive for mesenchymal stem cell [MSC] markers CD105, CD90, and CD73, and negative for HSC markers CD34 and CD45. Our data demonstrated the differentiation potential of ASCs to osteoblasts by alizarin red and alkaline phosphatase staining. MTT assay results showed a higher proliferation rate of CD34+cells directly cultured on the ADSCs feeder layer group compared to the other groups. Direct contact between HSCs and the feeder layer was prevented by a microporous membrane p53 expression increased in the HSCs group with indirect contact of the feeder layer compared to direct contact of the feeder layer. p53 significantly downregulated in HSCs cultured on ADSCs, whereas miR-145 significantly upregulated in HSCs cultured on ADSCs

Conclusion: ADSCs might increase HSCs proliferation and self-renewal through miR-145, p53, and their relationship

effects of iron oxide nanoparticle on differentiation of human mesenchymal stem cells to osteoblast

Introduction: Iron oxide nanoparticles [IO NP] have an increasing number of biomedical applications. To date, the potential cytotoxicity of these particles remains an issue of debate. Little is known about the cellular interaction or toxic effects of IO NP on differentiation of stem cells. The aim of the present study was to investigate the possible toxic role of different doses of IO NP in differentiation of human mesenchymal stem cells [hMSCs] derived bone marrow to osteoblast

Methods: hMSCs were seeded on normal stem cell medium with added 20 and 70microg/ml of IO NPs. Post confluence [2 weeks], cells growing cell viability was measured by 3-[4, 5-Dimethylthiazol-2-yl] -2, 5 -diphenyltetrazolium bromide [MTT] assays. hMSCs at passage 2 were cultured in osteogenic medium added with 20 and 70 microg/ml of IO NPs. The expression of osteogenesis markers of osteopontin, osteocalcine in different groups was compared by RT-PCR assay

Results: Our findings showed that cell viability of hMSCs cultured in normal stem cell media containing 20microg/ml IO NPs was significantly [P>0.05] lower than control and 70microg/ml dose of IO NPs groups, while there was no significant difference between control and 70microg/ml dose of IO NP. The expression level of osteoblast markers osteopontin and osteocalcin in hMSCs differentiated to osteoblast in dose with 20microg/ml IO NPs was significantly lower than the other groups, while the expression level of osteopontin and osteocalcin in 70microg/ml IO NPs dose was insignificantly higher than control group

Conclusion: To summarize, the presence of IO NPs with low dose influenced the cell viability cells in normal stem cell media, demonstrating toxicity of this material with 20 microg/ml. It could be probably due to penetrating particles throughout cell membrane. This represents a critical aspect to its successful use for stem cell-based regenerative medicine strategies

effects of zinc oxide nanoparticles on differentiation of human mesenchymal stem cells to osteoblast

The mesenchymal stem cells [MSCs] have been introduced as appropriate cells for tissue engineering and medical applications. Some studies have shown that topography of materials especially physical surface characteristics and particles size could enhance adhesion and proliferation of osteoblasts. In the present research, we studied the distinction effect of 30 and 60 microg/ml of zinc oxide [ZnO] on differentiation of human mesenchymal stem cells to osteoblast. After the third passage, human bone marrow mesenchymal stem cells were exposed to 30 and 60 microg/ml of ZnO nanoparticles having a size of 30 nm. The control group has received no ZnO nanoparticles. On day 15 of incubation for monitoring the cellular differentiation, alizarin red staining and RT-PCR assays were performed to evaluate the level of osteopontin, osteocalsin and alkaline phosphatase genes expression. In the group receiving 30 microg/ml of ZnO nanoparticles, the expression of osteogenic markers such as alkaline phosphatase, osteocalcin and osteopontin genes were significantly higher than both control and the group receiving 60 microg/ml ZnO nanoparticle. These data also confirmed by alizarin red staining. It seems the process of differentiation of MSCs affected by ZnO nanoparticles is dependent on dose as well as on the size of ZnO

Comparison of biocompatibility of various membranes with fibroblastsy

Objective: Different techniques have been suggested for the repair of bone defects at the injured sites. Use of biomembranes, or application of plasma rich in growth factor [PRGF] at the site of proliferation of osteoblasts are among the suggested techniques. The current study aimed to compare the biocompatibility of human periodontal ligament fibroblasts [hPLF] cultured on Hypro-Sorb F, Pericardium and Tutodent resorbable membranes coated with PRGF

Methods: This experimental study was conducted on four resorbable membranes namely Hypro- Sorb F, Pericardium, Tutodent and Vicryl. Fibroblast cells isolated from the periodontal ligament of premolar tooth were passaged three times and 10[5]cells/ cm[2] were cultured on membranes coated with PRGF. After 72 hours, the cells were evaluated in terms of biocompatibility and alkaline phosphatase [ALP] activity. Statistical analysis was carried out using one-way ANOVA

Results: PRGF increased cell adhesion and Tutodent membrane coated with PRGF showed the highest cell adhesion compared with Hypro-Sorb F and Pericardium membranes [p=0.005]

Conclusion: PRGF increases cell viability and ALP activity of cells on biomembranes. PRGF treatment increases the adhesion of fibroblast cells to these membranes

effect of carbon dioxide [CO[2]] laser irradiation with a power of 6w on sandblasting with large grit and acid etching [SLA] titanium discs through measuring sarcoma osteogenic [SaOS-2] human osteoblast-like cell viability

The purpose of this study was to investigate the effect of 6W power Carbon Dioxide Laser [CO[2]] on the biologic compatibility of the Sandblasting with large grit and acid etching [SLA] titanium discs through studying of the Sarcoma Osteogenic [SaOS-2] human osteoblast-like cells viability. Sterilized titanium discs were used together with SaOS-2 human osteoblast-like cells. 6 sterilized SLA titanium discs of the experimental group were exposed to irradiation by CO[2] laser with a power of 6W and 10.600nm wavelength, at fixed frequency of 80Hz during 45 seconds in both pulse and non-contact settings. SaOS-2 human osteoblast-like cells were incubated under 37 C in humid atmosphere [95% weather, 5% CO[2]] for 72 hours. MTT test was performed to measure the ratio level of cellular proliferation. The results indicated that at 570nm wavelength, the 6W CO[2] laser power have not affected the cellular viability. CO[2] laser in 6w power has had no effect on the biologic compatibility of the SLA titanium surface

Comparison of manual tools, ultrasonic and erbium-doped yttrium aluminum garnet [er:yag] laser on the debridement effect of the surface of the root of teeth suffering from periodontitis

Periodontal diseases are considered as some of the most common reasons of teeth loss, which occur due to the aggregation of microbial plaque and other precipitations on the dental surfaces. In this study, the scaling effect using manual tools, ultrasonic machine and Erbium-Doped Yttrium Aluminum Garnet [Er:YAG]laser on the connection of the human gums connective tissue cells on the root surface of the teeth suffering from severe periodontitis will be compared. After removal of the big precipitations with manual tools, Er:YAG laser light emission of Photona machine is used with respect to the following characteristics: wavelength: 2940 micro m, each pulse: 100mJ, frequency: 10 pulse/sec, optic fiber with cross section 0.5x1.65mm, fiber tip angle with root surface: 15-20 degrees with non-contact mode, 1.5mm farther than the root surface and pulse duration 230 very short. The gingival fibroblast cellular was incubated as a sample of the human gums connective tissue cells under 37C. These cells were departed from the culture medium after the cellular reproduction in the third passage. On the 3[rd] day after incubation, the gingival fibroblast cells morphology was studied by Scanning Electron Microscopy [SEM]. The results of SEM images in the present study indicated the spread fibroblast cells with philopodia were found in all of 5 groups; untreated healthy group [control], untreated group suffering from periodontitis, the scaling effect using manual tools [Scaled Gracey], ultrasonic machine and Er:YAG laser. There is a meaningful difference among the three treatment groups [P<0.001] in the numbers of the fibroblast cells, while all the four treated groups had a meaningful difference with the positive control group [P < 0.001]. The present study indicated that although various dental surfaces cleaning methods may be different in other aspects, but are similar concerning the fibroblasts morphology. Also in addition to power, laser emission time may also be effective in the cells morphology results.

Prenatal exposure to maternal voluntary exercise during pregnancy provides protection against mild chronic postnatal hypoxia in rat offspring

Postnatal hypoxia is a main cause of neuronal damage in newborn. However, our understanding of the possible preventive or therapeutic methods to reduce the harmful effects of hypoxia is still primary. Pregnant rats were provided with running wheels during their pregnancy. On PND4 [postnatal day 4] to PND8, the rat pups were exposed to postnatal chronic hypoxia [11% O[2], 89% N[2]] in an air-tight plastic chamber for a period of six hours per day. The number of neurons and also angiogenesis in hippocampus were studied. Postnatal exposure to mild hypoxia decreased the number of the neurons in all studied regions of the hippocampus CA1, CA3 [cornu ammonis], DG [dentate gyrus] and SUB [cubiculum] in rat pups. In other words the number of the neurons in rat pups born from voluntary exercise group was not significantly less than control group in CA1, CA3 and DG regions. So maternal Voluntary exercise during pregnancy increases the blood vessel density in the DG region of the hippocampus of the rat pups. In this study for the first time we provide evidences that show the protective effect of maternal voluntary exercise during pregnancy on rat offspring against postnatal hypoxia. We revealed that maternal exercise during pregnancy increases the hippocampal neuron number and angiogenesis in offspring

[Effect of biomaterial grafts of Totudent and Bio-oss on mitochondrial activity and alkaline phosphatase levels of Saos-2 osteoblastoid cells]

Several reports on clinical utility of various Bone Substitute Materials, [BSM] for grafting have shown successful results. The most important factor in successful grafting of BSM is the physiological and histological behavior of this material which can be evaluated by observing, rate of survival and proliferation of osteogenic cells. This research was conducted to examine the effect of Totudent and Bio-oss on the morphology, proliferation and rate of survival of Saos-2 osteoblastoid cells in vitro. This experimental study was done on two types of materials used as bone grafts viz. Totudent and Bio-oss. Twelve samples from each of these 2 materials grafted on Saos-2 osteoblastoid cells and 4 negative control samples were studied. On day 15, the rate of cell survival was determined by the MTT test and on day 20, Alkaline phosphatase activity of these cells in the culture medium were evaluated by the Alkaline phosphatase kit. The rate of cell survival in the Bio-oss and Tutodent group was significantly diminished in comparison with the control group. This rate was significantly higher in the cells grafted with Tutodent as compared to Bio-oss; alkaline phosphatase activity in Totudent group was higher than the other 2 groups. Tutodent bone graft showed better result in the morphologic variations of Saos-2 cells in the culture medium as compared to Bio-oss, and has more appropriate biologic characteristics

[Comparing bone marrow mesenchymal cells and mesothelial cells of peritoneal fluid in expressing molecules of major histocompatibility complex [MHC]]

Mesothelium is composed of a single layer of mesothelial cells attached to a thin basement membrane supported by subserosal connective tissue; it plays an important role in homeostasis, wound healing, fluid transport and inflammation. The introduction of peritoneal dialysis [PD] as a modality of renal replacement therapy has provoked much interest in the biology of peritoneal mesothelial cell. This study was conducted to investigate the nature and plasticity of mesothelial cells isolated from peritoneal dialysed fluid [PDF], from patients on dialysis. Cell populations from peritoneal fluid of dialysed patients showing no evidence of peritonitis, and mesenchymal cells isolated from the bone marrow were cultured and screened for dominant surface markers by cell culture and immunofluorescence; MHC II, [DR, DQ], and MHC I were detected by flow cytometry method. Our results showed both bone marrow mesenchymal stem cells and mesenchymal cells obtained from peritoneal fluid do not express MHC II; also, peritoneal mesothelial cells have limited expression of MHC I. The use of peritoneal mesothelium as an option for grafting and regenerative therapy needs to be investigated in intensive research

[Scanning electron microscopy study of morphology of totudent and bio-oss bone grafts on SaOS-2 cells]

Several reports have been published on clinical application of various Bone Substitute Material [BSM] which show their successful application. The most important factor in determination of BSM success is the physiological and histological behavior of this material that is evaluated by morphology of osteogenic cells, rate of survival and their proliferation. This research was done to assess the effect of this material on morphology, proliferation and rate of survival of SaOS-2 osteoblastoid cells. In this experimental in vitro study, two types of materials of bone graft [totudent and bio-oss] were assessed for their effect on survival rate, proliferation and morphology of SaOS-2 cells. 12 samples from each of grafted material and 4 negative control samples have been used. In day 21, morphology of these cells was evaluated by scanning electron microscopy [SEM]. In the term of spiral morphology, there was the meaningful statistics difference between Cerasorb and two other groups, but there was not meaningful difference between two groups of totudent and control. In the term of spindle morphology, there was meaningful statistics difference between the above mentioned groups. In the term of elongated morphology with pseudopodia, none of them had meaningful statistics difference. The synthetic material of totudent bone graft compared with bio-oss, showed better result in the morphologic variations of SaOS-2 cells at culture medium and has better biologic characteristic