scholarly journals The Osteogenic Potential of Human Nondifferentiated and Pre-differentiated Mesenchymal Stem Cells Combined with an Osteoconductive Scaffold – Early Stage Healing

2017 ◽  
Vol 60 (1) ◽  
pp. 12-18 ◽  
Author(s):  
Luboš Tuček ◽  
Zuzana Kočí ◽  
Kristýna Kárová ◽  
Helena Doležalová ◽  
Jakub Suchánek
Keyword(s):  
Stem Cells ◽  
Wistar Rats ◽  
Early Stage ◽  
Osteogenic Potential ◽  
Control Group ◽  
Positive Staining ◽  
Bone Scaffold ◽  
Synthetic Bone ◽  
Differentiated Cells ◽  
The Right

Despite the huge research into stem cells and their regenerative properties for bone healing, there are still unanswered questions including the recipient’s respond to the presence of the stem cells, the fate of stem cells inside the bone defect and the possible advantage in utilizing pre-differentiated cells. To address these problems, we used human multipotent mesenchymal stromal/stem cells (MSCs), GMP Grade, in a rat model of bone formation. In a “bioreactor concept” approach seven Wistar rats were implanted with 0.2 g of synthetic bone scaffold seeded with 2 × 106 MSCs, seven Wistar rats were implanted with 0.2 g of synthetic bone scaffold seeded with 1 × 106 predifferentiated osteoblasts and 1 × 106 pre-differentiated endothelial cells and 14 Wistar rats were implanted with 0.2 g of synthetic bone scaffold without seeded cells into an intramuscular pocket on the left side of their back. The right side of each rat was used as a control, and 0.2 g of synthetic bone scaffold was implanted into the intramuscular pocket alone. To see the early stage healing the samples were harvested 14 days after the implantation, MSCs were detected by positive DAPI and MTCO2 staining in 43% of all the samples implanted with MSCs, and no inflammation signs were present in any implanted animal. New vessels could be found in both groups implanted with MSCs, but not in the control group of animals. However, hematoxylin-eosin staining could not detect newly created bone within the implant in any of the groups. These results were in line with COLL1 staining, where we could detect positive staining only in three cases, all of which were implanted with un-differentiated MSCs. According to our findings, there were no benefits of using the pre-differentiated of MSC.

scholarly journals Comparative evaluation of osteogenic differentiation potential of stem cells derived from dental pulp and exfoliated deciduous teeth cultured over granular hydroxyapatite based scaffold

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Manal Nabil Hagar ◽  
Farinawati Yazid ◽  
Nur Atmaliya Luchman ◽  
Shahrul Hisham Zainal Ariffin ◽  
Rohaya Megat Abdul Wahab
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
3D Culture ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Osteogenic Potential ◽  
Control Group ◽  
Rt Pcr ◽  
Differentiation Potential

Abstract Background Mesenchymal stem cells isolated from the dental pulp of primary and permanent teeth can be differentiated into different cell types including osteoblasts. This study was conducted to compare the morphology and osteogenic potential of stem cells from exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSC) in granular hydroxyapatite scaffold (gHA). Preosteoblast cells (MC3T3-E1) were used as a control group. Methodology The expression of stemness markers for DPSC and SHED was evaluated using reverse transcriptase-polymerase chain reaction (RT-PCR). Alkaline phosphatase assay was used to compare the osteoblastic differentiation of these cells (2D culture). Then, cells were seeded on the scaffold and incubated for 21 days. Morphology assessment using field emission scanning electron microscopy (FESEM) was done while osteogenic differentiation was detected using ALP assay (3D culture). Results The morphology of cells was mononucleated, fibroblast-like shaped cells with extended cytoplasmic projection. In RT-PCR study, DPSC and SHED expressed GAPDH, CD73, CD105, and CD146 while negatively expressed CD11b, CD34 and CD45. FESEM results showed that by day 21, dental stem cells have a round like morphology which is the morphology of osteoblast as compared to day 7. The osteogenic potential using ALP assay was significantly increased (p < 0.01) in SHED as compared to DPSC and MC3T3-E1 in 2D and 3D cultures. Conclusion gHA scaffold is an optimal scaffold as it induced osteogenesis in vitro. Besides, SHED had the highest osteogenic potential making them a preferred candidate for tissue engineering in comparison with DPSC.

scholarly journals Effect of Placental Extract on Omentum Mesenchymal Stem Cells Differentiation in NMRI Mice

2017 ◽  
Vol 7 (1) ◽  
pp. 176
Author(s):  
Maryam Sadat Nezhadfazel ◽  
Kazem Parivar ◽  
Nasim Hayati Roodbari ◽  
Mitra Heydari Nasrabadi
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Flow Cytometry ◽  
Endothelial Cells ◽  
Mesenchymal Stem Cells ◽  
Control Group ◽  
Fat Cell ◽  
Nmri Mice ◽  
Differentiated Cells ◽  
Placental Extract

Omentum mesenchymal stem cells (OMSCs) could be induced to differentiate into cell varieties under certain conditions. We studied differentiation of OMSCs induced by using placenta extract in NMRI mice. Mesenchymal stem cells (MSCs) were isolated from omentum and cultured with mice placenta extract. MSCs, were assessed after three passages by flow cytometry for CD90, CD44, CD73, CD105, CD34 markers and were recognized their ability to differentiate into bone and fat cell lines. Placenta extract dose was determined with IC50 test then OMSCs were cultured in DMEM and 20% placenta extract.The cell cycle was checked. OMSCs were assayed on 21 days after culture and differentiated cells were determined by flow cytometry and again processed for flow cytometry. CD90, CD44, CD73, CD105 markers were not expressed, only CD34 was their marker. OMSCs were morphologically observed. Differentiated cells are similar to the endothelial cells. Therefore, to identify differentiated cells, CD31 and FLK1 expression were measured. This was confirmed by its expression. G1 phase of the cell cycle shows that OMSCs compared to the control group, were in the differentiation phase. The reason for the differentiation of MSCs into endothelial cells was the sign of presence of VEGF factor in the medium too high value of as a VEGF secreting source.

scholarly journals Analysis of the muscle tissue of Wistar rats submitted to the sciatic nerve compression model and cryotherapy

2018 ◽  
Vol 16 (3) ◽  
Author(s):  
Jhenifer Karvat ◽  
Camila Mayumi Martin Kakihata ◽  
Lizyana Vieira ◽  
José Luis da Conceição Silva ◽  
Lucinéia de Fátima Chasko Ribeiro ◽  
...  
Keyword(s):  
Sciatic Nerve ◽  
Muscle Tissue ◽  
Wistar Rats ◽  
Nerve Compression ◽  
Control Group ◽  
Functional Evaluation ◽  
Cross Sectional ◽  
Injury Group ◽  
Significant Difference ◽  
The Right

ABSTRACT Objective: To evaluate the effects of right sciatic nerve compression and cryotherapy on muscle tissue. Methods: We used 42 male Wistar rats, subdivided in the following Groups Control, Injury 3, Injury 8 and Injury 15 submitted to nerve compression and euthanized in the 3rd, 8th and 15th day after surgery. The Cryotherapy Injury 3 was entailed treatment with cryotherapy by immersion of the animal in recipient for 20 minutes during 1 day, then animals were euthanized at the 3rd day after surgery, and the Cryotherapy Injury 8 and the Cryotherapy Injury 15 was treated for 6 days, and euthanized at the 8th and 15th day after surgery. Functional evaluation was performed by the grasping strength of the right pelvic limb. The right tibialis anterior muscles were evaluated for mass, smaller diameter and cross-sectional area. In the Cryotherapy Injury 8 and the Cryotherapy Injury 15 groups, the hydroxyproline was dosed in the right soles. Results: In the compression there was a significant difference in the Injury Groups compared with the Control Group (p<0.05). In the smaller diameter, the compression in Control Group was higher than Injury 8 (p=0.0094), Injury 15 (p=0.002) and Cryotherapy Injury 15 (p<0.001) groups. The comparison between groups with euthanasia in the same post-operative period, a significant difference (p=0.0363) was seen in day 8th after surgery, and this result in Cryotherapy Injury Group was greater than Injury Group. In the fiber area, Control Group was also higher than the Injury 8 (p=0.0018), the Injury 15 (p<0.001) and the Cryotherapy Injury 15 (p<0.001). In hydroxyproline, no significant difference was seen between groups. Conclusion: Nerve damage resulted in decreased muscle strength and trophism, the cryotherapy delayed hypotrophy, but this effect did not persist after cessation of treatment.

scholarly journals A Comparative In Vitro Analysis of the Osteogenic Potential of Human Dental Pulp Stem Cells Using Various Differentiation Conditions

2020 ◽  
Vol 21 (7) ◽  
pp. 2280 ◽  
Author(s):  
Terezia Okajcekova ◽  
Jan Strnadel ◽  
Michal Pokusa ◽  
Romana Zahumenska ◽  
Maria Janickova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Osteogenic Potential ◽  
Surface Markers ◽  
Osteogenic Medium ◽  
Differentiated Cells ◽  
Mineralization Potential ◽  
Gene And Protein Expression

Dental pulp stem cells (DPSCs) have excellent proliferative properties, mineralization potential and can be easily obtained from third molar teeth. Recently, many studies have focused on isolation and differentiation of DPSCs. In our study, we focused on biological properties of non-differentiated DPSCs in comparison with osteogenic differentiated cells from DPSCs. We analyzed morphology as well as mineralization potential using three varied osteogenic differentiation media. After fifteen days of differentiation, calcium deposit production was observed in all three osteogenic differentiation media. However, only one osteogenic medium, without animal serum supplement, showed rapid and strong calcification—OsteoMAX-XF™ Differentiation Medium. Therefore, we examined specific surface markers, and gene and protein expression of cells differentiated in this osteogenic medium, and compared them to non-differentiated DPSCs. We proved a decrease in expression of CD9 and CD90 mesenchymal stem cell surface markers, as well as downregulation in the expression of pluripotency genes (NANOG and OCT-4) and increased levels of expression in osteogenic genes (ALP, BSP, OCN and RUNX2). Moreover, osteogenic proteins, such as BSP and OCN, were only produced in differentiated cells. Our findings confirm that carefully selected differentiation conditions for stem cells are essential for their translation into future clinical applications.

Bone Marrow Derived Mesenchymal Stem Cell Augmentation of Rabbit Flexor Tendon Healing

Hand Surgery ◽  
2015 ◽  
Vol 20 (03) ◽  
pp. 421-429 ◽  
Author(s):  
Min He ◽  
Aaron Wei Tat Gan ◽  
Aymeric Yu Tang Lim ◽  
James Cho Hong Goh ◽  
James Hoi Po Hui ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Flexor Tendon ◽  
Tendon Repair ◽  
Tendon Healing ◽  
Control Group ◽  
High Dose ◽  
Positive Staining ◽  
Flexor Tendon Repair

Background: This study investigated the effect of mesenchymal stem cell implantation on flexor tendon healing using a rabbit model of flexor tendon repair. Specifically, we compared the difference between autologous and allogeneic stem cells. The influence of cell number on the outcome of flexor tendon healing was also investigated. Methods: Repaired tendons on the rear paws of rabbits were randomly assigned into four groups: control group, 1 million autologous cells, 1 million allogeneic cells, and 4 million allogeneic cells. Rabbits were sacrificed at 3 or 8 weeks after surgery. Results: Implantation of 4 million stem cells resulted in a significant increase in range of motion compared with control group at three weeks after surgery. The positive staining of collagen I in healing tendons was enhanced in stem cell treated groups three weeks after surgery. However, stem cells did not improve biomechanical properties of flexor tendons. Conclusions: High dose stem cells attenuated adhesions in the early time point following flexor tendon repair. Further work is needed determine the value of stem cell therapy in flexor tendon healing in humans.

scholarly journals Modulation of gene transcription promoted by mesenchymal stem cells on cation-chloride cotransporter NKCC1 in experimental epilepsy

2021 ◽  
Author(s):  
Giovani Zocche Junior ◽  
Isadora Ghilardi ◽  
Laura Provenzi ◽  
Gabriel Leal ◽  
Giulia Pinzetta ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Wistar Rats ◽  
Brain Structures ◽  
Treated Group ◽  
Control Group ◽  
Neural Firing ◽  
Post Transplantation ◽  
Neuroprotective Effects ◽  
Invasive Approach

Introduction: temporal lobe epilepsy is a disorder in which synchronized and rhythmic neural firing causes spontaneous recurrent seizures (1). Refractoriness due to this condition reaches 30% of its carriers (2,3). The search for therapeutic alternatives to help cope with this disease are extremely important. Mesenchymal stem cells (MSCs) appear as a plausible treatment option, as they present a less invasive approach and due to their niche modulating character (4,5). Objectives: this study aimed to quantify the gene expression of cation-chloride cotransporter NKCC1 encoded by the SLC12A2 gene in the encephalic tissue of pilocarpine-induced epileptic rats (6,7). Design: experimental study, brain institute of Rio Grande do Sul. Methods: MSCs were obtained from the bone marrow of Wistar rats, cultured, and transplanted through intravenous injection into control and epileptic Wistar rats. The rats were divided between control group, MSCs treated group, and pilocarpine group, containing 8 individuals each (8). Expression analysis was performed using real-time polymerase chain reaction. Results: for both 1 day and 7 days post-transplantation, an increase in the NKCC1 expression in both control and epileptic treated groups as compared to its expression in untreated epileptic and control groups with special attention to the amygdala, the hippocampus and the prefrontal cortex. Conclusion: MSCs stimulated expression of NKCC1 in brain structures of rats induced by pilocarpine to epilepsy. This corroborates the hypothesis of neuroprotective effects and modulating properties of stem cells and may point to more mechanisms for investigating the functioning and collaboration of these cells as a treatment for epilepsy.

scholarly journals Animal models of ovariectomy in rats demonstrate characteristics similar to early stage osteoarthritis

2021 ◽  
Author(s):  
Natália Aparecida Casonato ◽  
Camila Marques de Araújo ◽  
Mariane Santos Trevisan ◽  
Cristina Arrais Lima ◽  
Fernando Augusto Vasilceac
Keyword(s):  
Animal Models ◽  
Knee Joint ◽  
Wistar Rats ◽  
Early Stage ◽  
Experimental Period ◽  
Histological Evaluation ◽  
Control Group ◽  
Intermediate Region ◽  
Histological Changes ◽  
Deep Region

Abstract Purpose The aim of our study is to analyze the model of ovariectomy (OVX) in rats reproduced histological changes of osteoarthritis (OA). Methods For the development of the research, 12 Wistar rats were used, divided into 2 equal groups: Control Group - C (n = 6) and Osteoarthritis Group - OA (n = 6). After the 6-month experimental period, all rats were sacrificed and, subsequently, the entire knee joint complex was removed without disarticulation. For the histological evaluation of the tissue, the recommendations of the International Society for Research in OA (OARSI) were used. For data processing, each evaluation was statistically treated in both groups, comparing data from group C with the group OA. Results: Through the histological evaluation of OARSI, the evolution of OA in various tissues of the joint was evaluated. Although the OA group showed noticeable differences from group C, they were not as significant. Thus, only statistically significant favors were presented in the loss of the cartilaginous matrix (OA and C, p = 0.51), considering that the changes in the loss of ECM occurred only at the depth of 0% (superficial region), but at the depth of 50% (intermediate region) and 100% depth (deep region) did not exist. Conclusion Our study demonstrated that the OVX model is a good model to discuss OA, showing histological changes similar to those found in OA, the model demonstrated to have a progressive and slow characteristic since after the OARSI evaluations, prominent evidence was found in the initial manifestations of OA.

Abstract 188: Differentiation of Preexisting Cardiosphere-Derived Stem Cells Was Modulated by Telocyte Niche

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Xi-Yong Yu ◽  
Yu-Kai Huang ◽  
Yu Pan ◽  
Xiao-Hong Li ◽  
Qiu-Xiong Lin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Early Stage ◽  
Scar Tissue ◽  
Neonatal Rat ◽  
Control Group ◽  
Cardiac Progenitor Cells ◽  
Stem Cell Markers ◽  
Adult Rat ◽  
Sd Rats ◽  
Differentiation Efficiency

Background: Cardiosphere-derived stem cells (CSCs) were found to be unusual pre-existing cells, comparing with other kinds of stem cells, showing the greatest cardiomyocytes differentiation potency. This study is to explore whether the microenvironment provided by telocytes can improve differentiation efficiency of CSCs. Methods and Results: The CSCs and telocytes were isolated from myocardium of SD rats. The experiment was divided into control group, T-group (CSCs co-culture with adult rat telocytes), N-group (CSCs co-culture with neonatal rat caridomyocytes). Q-PCR was used to check the change of stem cell markers and cardiomyocyte markers. Immunofluorescence was used to detect the expression of cTnI protein in each group. The expression of Isl-1 in T-group was higher than other groups at 3 days, Nanong was higher at 5d, Gata4 and cTnI were higher at 9d. There are almost no expressions of cTnI protein in control group and N-group, but higher expressions of cTnI protein in T-group. The data of PCR-array showed that the most biomarkers of stem cells increased when CSCs co-culture with telocytes for 3 days, but the most biomarkers of cardiac progenitor cells increased when CSCs co-culture with telocytes for over 9 days. Finally, the CSCs induced by telocytes (iCSCs) were injected into borders of cardiac scar tissue 1 wk after experimental infarction. The results showed a nearly normal ejection fraction (EF) in iCSCs-treated rats but a lower EF in all PBS-treated animals. Conclusion: There are bilateral regulations of the microenvironment provided by telocytes which not only maintain the stemness of CSCs at the early stage, but also improve the differentiation efficiency of CSCs at the late stage. Telocyte is an important chaperone of CSCs, and helps for repairing damaged heart muscle. This work was supported by NSFC (81330007) and 973 program of China (2012CB526600).

Association of Dental Pulp Stem Cells and Ricinus Bone Compound in a Model of Bone Defect

2020 ◽  
Vol 3 (3) ◽  
pp. 267-278
Author(s):  
Alan Jesus ◽  
Adriano Jesus ◽  
Flávia Lima ◽  
Luiz Freitas ◽  
Cássio Meira ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Bone Defect ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Osteogenic Potential ◽  
Autogenous Bone ◽  
Control Group

Autogenous bone grafting is needed in some bone tissue defects; however, it causes secondary surgical wounds and morbidity. Tissue bioengineering may be an alternative approach for bone regeneration. Here we investigated the osteogenic potential of dental pulp stem cells from deciduous teeth (DPSC) in association with a Ricinus bone compound (RBC) in a model of bone defect. The influence of the biomaterial RBC on the proliferation and osteogenic differentiation of DPSC was assessed in vitro by MTT metabolism and alizarin red staining, respectively. The morphologic analysis was performed using the optic and scanning electron (SEM) microscopies. For the in vivo study, 54 Wistar rats submitted to calvarial defects were filled with RBC or RBC+DPSC. A control group had the defects filled only with blood clots. Analyses were performed 15, 30 and 60 days after treatment using digital radiography, optical microscopy, SEM and chemical analysis by electron dispersive spectroscopy. The Ricinus bone compound (RBC) did not inhibit the osteogenic differentiation in vitro. No spontaneous regeneration was observed in the control group. The area of the calvarial defect of the RBC+DPSC group showed greater radiopacity on day 15. The RBC presented no reabsorption, was biocompatible and showed osteointegration, working as a mechanical filling. Only sparse ossification areas were found and those were larger and more developed on the RBC+DPSC group when compared to animals treated only with RBC. RBC in association with DPSC is a promising combination for applications in bone regeneration.  

Role of E1B protein-dificient oncolytic virus in breast cancer stem cells

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e22131-e22131
Author(s):  
F. Zhang ◽  
Y. Ma ◽  
Y. Xu ◽  
M. Huang ◽  
C. Song ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Oncolytic Adenovirus ◽  
Infected Group ◽  
Control Group ◽  
Breast Cancer Stem Cells ◽  
Differentiated Cells ◽  
Mcf 7

e22131 Background: Cancer stem cells have been indicated in the initiation of tumors and are even found to be responsible for relapses after apparently curative therapies have been undertaken. In breast cancer, they may reside in the CD44+CD24−/low population. Oncolytic adenoviruses enter cells through infection and can kill both proliferating and quiescent cells. We investigated the role of E1B protein- dificient oncolytic adenovirus in breast cancer stem cells. Methods: MCF-7 cells were infected by E1B protein-dificient oncolytic adenovirus as infected group (MOI=100) and cultured routinely as control group simultaneously. The proportion of CD44+CD24- cells was assessed by flow cytometry (FCM) in two groups respectively. Meanwhile, mammosphere culture was done in two groups' cells to observe the size and number of mammospheres, calculate the mammosphere- forming efficiency (MFE). The proportion of CD44+CD24- cells in two groups' mammospheres was assessed by FCM. Results: The percentages of CD24-,CD44+, CD44+CD24- in the infected gruop were 43.9%, 63.26%, 22.19%, respectively. While in the control group, the percentages were 6.74%, 88.30%, 2.30%. In the infected group, the time of mammosphere's formation was earlier, the volumes of mammospheres were bigger and the MFE was higher than the control group (1.26%:0.9%). In two mammospheres' groups, the proportion of CD44+CD24- cells in experiment group and control group was 38.08% and 23.35%, respectively. Conclusions: E1B protein-dificient oncolytic adenovirus can kill MCF-7 cells in short time, mainly breast cancer differentiated cells. It maybe promote the growth of the breast cancer stem cells. It maybe accelerate the speed of self-renewed and differentiation of the breast cancer stem cells. No significant financial relationships to disclose.

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