scholarly journals Bone remodeling using a three-dimensional chitosan - hydroxyapatite scaffold seeded with hypoxic conditioned human amnion mesenchymal stem cells

2021 ◽  
Vol 54 (2) ◽  
pp. 68
Author(s):  
Michael Josef Kridanto Kamadjaja
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Remodeling ◽  
Three Dimensional ◽  
Wistar Rat ◽  
Control Group ◽  
Human Amnion ◽  
Morphogenetic Protein ◽  
Hydroxyapatite Scaffold ◽  
Endothelial Growth Factor

Background: Bone regeneration studies involving the use of chitosan–hydroxyapatite (Ch-HA) scaffold seeded with human amnion mesenchymal stem cells (hAMSCs) have largely incorporated tissue engineering experiments. However, at the time of writing, the results of such investigations remain unclear. Purpose: The aim of this study was to determine the osteogenic differentiation of the scaffold Ch-HA that is seeded with hAMSCs in the regeneration of calvaria bone defect. Methods: Ch-HA scaffold of 5 mm diameter and 2 mm height was created by lyophilisation and desalination method. hAMSCs were cultured in hypoxia environment (5% oxygen, 10% carbon dioxide, 15% nitrogen) and seeded on the scaffold. Twenty male Wistar rat subjects (8 – 10 weeks, 200 - 250 grams) were randomly divided into two groups: control and hydroxyapatite scaffold (HAS). Defects (similar size to scaffold size) were created in the calvaria bone of the all-group subjects, but a scaffold was subsequently implanted only in the treatment group members. Control group left without treatment. After observation lasting 1 and 8 weeks, the subjects were examined histologically and immunohistochemically. Statistical analysis was done using ANOVA test. Results: Angiogenesis; expression of vascular endothelial growth factor; bone morphogenetic protein; RunX-2; alkaline phosphatase; type-1 collagen; osteocalcin and the area of new trabecular bone were all significantly greater in the HAS group compared to the control group. Conclusion: The three-dimensional Ch-HA scaffold seeded with hypoxic hAMSCs induced bone remodeling in calvaria defect according to the expression of the osteogenic and angiogenic marker.

scholarly journals Comparison of Immunosuppressive and Angiogenic Properties of Human Amnion-Derived Mesenchymal Stem Cells between 2D and 3D Culture Systems

2019 ◽  
Vol 2019 ◽  
pp. 1-16 ◽  
Author(s):  
Vitale Miceli ◽  
Mariangela Pampalone ◽  
Serena Vella ◽  
Anna Paola Carreca ◽  
Giandomenico Amico ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Three Dimensional ◽  
3D Culture ◽  
Culture Method ◽  
Paracrine Factors ◽  
Human Amnion ◽  
Tissue Repair And Regeneration ◽  
3D Culture System

The secretion of potential therapeutic factors by mesenchymal stem cells (MSCs) has aroused much interest given the benefits that it can bring in the field of regenerative medicine. Indeed, the in vitro multipotency of these cells and the secretive capacity of both angiogenic and immunomodulatory factors suggest a role in tissue repair and regeneration. However, during culture, MSCs rapidly lose the expression of key transcription factors associated with multipotency and self-renewal, as well as the ability to produce functional paracrine factors. In our study, we show that a three-dimensional (3D) culture method is effective to induce MSC spheroid formation, to maintain the multipotency and to improve the paracrine activity of a specific population of human amnion-derived MSCs (hAMSCs). The regenerative potential of both 3D culture-derived conditioned medium (3D CM) and their exosomes (EXO) was assessed against 2D culture products. In particular, tubulogenesis assays revealed increased capillary maturation in the presence of 3D CM compared with both 2D CM and 2D EXO. Furthermore, 3D CM had a greater effect on inhibition of PBMC proliferation than both 2D CM and 2D EXO. To support this data, hAMSC spheroids kept in our 3D culture system remained viable and multipotent and secreted considerable amounts of both angiogenic and immunosuppressive factors, which were detected at lower levels in 2D cultures. This work reveals the placenta as an important source of MSCs that can be used for eventual clinical applications as cell-free therapies.

scholarly journals Synergistic Effects of Orbital Shear Stress onIn VitroGrowth and Osteogenic Differentiation of Human Alveolar Bone-Derived Mesenchymal Stem Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-18 ◽  
Author(s):  
Ki Taek Lim ◽  
Jin Hexiu ◽  
Jangho Kim ◽  
Hoon Seonwoo ◽  
Pill-Hoon Choung ◽  
...  
Keyword(s):  
Stem Cells ◽  
Shear Stress ◽  
Mesenchymal Stem Cells ◽  
Alveolar Bone ◽  
Synergistic Effects ◽  
Osteoblastic Differentiation ◽  
Bone Morphogenetic Protein 2 ◽  
Control Group ◽  
Protein Levels ◽  
Morphogenetic Protein

Cellular behavior is dependent on a variety of physical cues required for normal tissue function. In order to mimic native tissue environments, human alveolar bone-derived mesenchymal stem cells (hABMSCs) were exposed to orbital shear stress (OSS) in a low-speed orbital shaker. The synergistic effects of OSS on proliferation and differentiation of hABMSCs were investigated. In particular, we induced the osteoblastic differentiation of hABMSCs cultured in the absence of OM by exposing hABMSCs to OSS (0.86–1.51 dyne/cm2). Activation of Cx43 was associated with exposure of hABMSCs to OSS. The viability of cells stimulated for 10, 30, 60, 120, and 180 min/day increased by approximately 10% compared with that of control. The OSS groups with stimulation of 10, 30, and 60 min/day had more intense mineralized nodules compared with the control group. In quantification of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2) protein, VEGF protein levels under stimulation for 10, 60, and 180 min/day and BMP-2 levels under stimulation for 60, 120, and 180 min/day were significantly different compared with those of the control. In conclusion, the results indicated that exposing hABMSCs to OSS enhanced their differentiation and maturation.

scholarly journals Intraarticular Allogenic Mesenchymal Stem Cells and Vascular Endothelial Growth Factor Injection in Anterior Cruciate Ligament Reconstruction

2020 ◽  
Vol 56 (1) ◽  
pp. 41
Author(s):  
Dwikora Novembri Utomo ◽  
Ferdiansyah Mahyudin ◽  
Arif Zulkarnain ◽  
Purwati Purwati ◽  
Rossy Setyawati
Keyword(s):  
Stem Cells ◽  
Vascular Endothelial Growth Factor ◽  
Mesenchymal Stem Cells ◽  
Anterior Cruciate Ligament ◽  
Cruciate Ligament ◽  
Hamstring Tendon ◽  
Control Group ◽  
Vascular Endothelial ◽  
Anterior Cruciate ◽  
Endothelial Growth Factor

Graft tunnel healing is important for the successful reconstruction of the anterior cruciate ligament by using the hamstring tendon autograft. There are studies that intra graft tunnel Bone Marrow Mesenchymal stem cells (BMSCs) or intra graft tunnel Vascular endothelial growth factor (VEGF) accelerated graft tunnel healing. This study aimed to investigate the effect of using BMSCs+VEGF injected intra-articular on graft tunnel healing. We reconstructed the anterior cruciate ligament (ACL) of 12 rabbits using an autograft hamstring tendon with and without intra-articular BMSCs+VEGF. Histological evaluation was done at 3 and 6 weeks after ACL reconstruction. On the surface between the graft and the bone tunnel obtained collagen fiber thickness or Sharpey fiber is significantly more than the control group (p< 0.05) in the evaluation of 3 weeks and 6 weeks either side of the tibia and the femur. To evaluate the progression of the treatment, treatment group and control group gained progression had significantly when compared to 3 weeks and 6 weeks. It can be concluded that intra-articular injection of VEGF+BMSCs can accelerate the integration of the graft tunnel from histology evaluation on 3 and 6 weeks.

scholarly journals Effects of icariin on the proliferation and osteogenic differentiation of human amniotic mesenchymal stem cells

2020 ◽  
Vol 15 (1) ◽  
Author(s):  
Fang Wang ◽  
Zhiyong Yang ◽  
Wei He ◽  
Qinggao Song ◽  
Kun Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Immunocytochemical Staining ◽  
Control Group ◽  
Engineering Technology ◽  
Alizarin Red ◽  
Human Amnion ◽  
Alp Activity ◽  
Amniotic Mesenchymal Stem Cells

Abstract Background Tissue engineering technology has been applied extensively for clinical research and human amnion mesenchymal stem cells (hAMSCs) could cause mesenchymal stem cells to differentiate into the bone tissue. However, it is necessary to develop and identify the safer appropriate amount of osteogenic inducer. The objective of this study is to investigate the effect of icariin (ICA) on the proliferation and osteogenic differentiation of hAMSCs. Methods The morphology and phenotype of hAMSCs were discovered by flow cytometry and immunocytochemical staining. The osteogenic differentiation of hAMSCs under the influence of different concentrations of ICA were assessed by alkaline phosphatase (ALP) activity substrate assay and alizarin red staining. Results MTT assay revealed that the hAMSCs pretreated with ICA exhibited increased proliferation when compared with the control group, and the most optimum concentration of ICA was 1 × 10− 6 mol/L. The combined analysis of ALP activity and ARS staining showed that ICA could significantly promote the osteogenic differentiation of hAMSCs, and the effect was most significant when the concentration of ICA was 1 × 10− 6 mol/L. Conclusion All the above results implied that ICA could significantly increase proliferation and enhance the osteogenic differentiation of hAMSCs, especially when the concentration of ICA was 1 × 10− 6 mol/L.

scholarly journals Effects of icariin on the proliferation and osteogenic differentiation of human amniotic mesenchymal stem cells

2020 ◽  
Author(s):  
Fang Wang ◽  
Zhiyong Yang ◽  
He Wei ◽  
Qinggao Gao ◽  
Kun Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Immunocytochemical Staining ◽  
Control Group ◽  
Engineering Technology ◽  
Alizarin Red ◽  
Human Amnion ◽  
Alp Activity ◽  
Amniotic Mesenchymal Stem Cells

Abstract Background Tissue engineering technology has been applied extensively for clinical research and human amnion mesenchymal stem cells (hAMSCs) could cause mesenchymal stem cells to differentiate into the bone tissue. However, it is necessary to develop and identify the safer appropriate amount of osteogenic inducer. The objective of this study is to investigate the effect of icariin (ICA) on the proliferation and osteogenic differentiation of hAMSCs. Methods The morphology and phenotype of hAMSCs were discovered by flow cytometry and immunocytochemical staining. The osteogenic differentiation of hAMSCs under the influence of different concentrations of ICA were assessed by Alkaline phosphatase (ALP) activity substrate assay and Alizarin red Staining. Results MTT assay revealed that the hAMSCs pretreated with ICA exhibited increased proliferation when compared with the control group, and the most optimum concentration of ICA was 1 × 10− 6 mol/L. The combined analysis of ALP activity and ARS staining showed that ICA could significantly promote the osteogenic differentiation of hAMSCs, and the effect was most significant when the concentration of ICA was 1 × 10− 6 mol/L. Conclusion All above results implied that ICA could significantly increase proliferation and enhanced the osteogenic differentiation of hAMSCs, especially when the concentration of ICA was 1 × 10− 6 mol/L.

scholarly journals Intraarticular Allogenic Mesenchymal Stem Cells and Vascular Endothelial Growth Factor Injection in Anterior Cruciate Ligament Reconstruction

2021 ◽  
Vol 56 (1) ◽  
pp. 41
Author(s):  
Dwikora Novembri Utomo ◽  
Ferdiansyah Mahyudin ◽  
Arif Zulkarnain ◽  
Purwati Purwati ◽  
Rossy Setyawati
Keyword(s):  
Stem Cells ◽  
Vascular Endothelial Growth Factor ◽  
Mesenchymal Stem Cells ◽  
Anterior Cruciate Ligament ◽  
Cruciate Ligament ◽  
Hamstring Tendon ◽  
Control Group ◽  
Vascular Endothelial ◽  
Anterior Cruciate ◽  
Endothelial Growth Factor

Graft tunnel healing is important for the successful reconstruction of the anterior cruciate ligament by using the hamstring tendon autograft. There are studies that intra graft tunnel Bone Marrow Mesenchymal stem cells (BMSCs) or intra graft tunnel Vascular endothelial growth factor (VEGF) accelerated graft tunnel healing. This study aimed to investigate the effect of using BMSCs+VEGF injected intra-articular on graft tunnel healing. We reconstructed the anterior cruciate ligament (ACL) of 12 rabbits using an autograft hamstring tendon with and without intra-articular BMSCs+VEGF. Histological evaluation was done at 3 and 6 weeks after ACL reconstruction. On the surface between the graft and the bone tunnel obtained collagen fiber thickness or Sharpey fiber is significantly more than the control group (p< 0.05) in the evaluation of 3 weeks and 6 weeks either side of the tibia and the femur. To evaluate the progression of the treatment, treatment group and control group gained progression had significantly when compared to 3 weeks and 6 weeks. It can be concluded that intra-articular injection of VEGF+BMSCs can accelerate the integration of the graft tunnel from histology evaluation on 3 and 6 weeks.

Prospects for the use of fibrin scaffolds populated with pulp and periodontal stem cells: an experimental study

2021 ◽  
Vol 26 (2) ◽  
pp. 96-103
Author(s):  
Yu. A. Dombrovskaya ◽  
N. I. Enukashvili ◽  
R. E. Banashkov ◽  
N. Yu. Semenova ◽  
I. A. Karabak ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Remodeling ◽  
Fibrin Glue ◽  
Research Work ◽  
Three Dimensional ◽  
Control Group ◽  
Laboratory Mice ◽  
The North ◽  
Promising Tool ◽  
Defect Area

Relevance. Creating three-dimensional scaffolds from biodegradable materials and seeding them with stem cells derived from the oral tissues is a promising tool for guided tissue regeneration. Pulp and periodontal stem cells have a high potential for osteogenic differentiation, which biologically determines their use in surgical bone reconstruction. The experiment shows the result of using fibrin glue seeded with pulp and periodontal stem cells on the mandible of laboratory mice. The article presents the results of computed tomography and histological examination. The data provide evidence of the influence of seeded scaffolds on bone remodeling in the area of the defect.Materials and methods. The Local Ethics Committee of the North-Western State Medical University named after I.I. Mechnikov gave permission for the practical part of the research work. The study included 29 white laboratory mice. Molars were extracted and a bone defect was formed. Pulp and periodontal stem cells were obtained and cell-seeded scaffolds were made, then they were introduced into the defect area. The animals were euthanized, maxillofacial CT scan and histology of the defect area were performed 28 days after the molar extraction.Results. The oral cavity of mice was examined, molars were extracted, and teeth were morphologically examined under anesthesia. Scaffolds were synthesized and bone defects were filled. CT scans and histology results were analyzed. The bone volume increased in the main group compared to the control group.Conclusion. The fibrin glue can be used to obtain a material with mechanical characteristics sufficient for a stable shape scaffold. The study proved that the pulp stem cells enclosed in a fibrin glue-based scaffold can maintain the ability to proliferate and osteogenically differentiate. The scaffold based on fibrin glue, which we used, affected the bone remodeling process in the area of jaw defects. 

scholarly journals Effects of icariin on the proliferation and osteogenic differentiation of human amniotic mesenchymal stem cells

2020 ◽  
Author(s):  
Fang Wang ◽  
Zhiyong Yang ◽  
He Wei ◽  
Qinggao Gao ◽  
Kun Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Immunocytochemical Staining ◽  
Control Group ◽  
Engineering Technology ◽  
Alizarin Red ◽  
Human Amnion ◽  
Alp Activity ◽  
Amniotic Mesenchymal Stem Cells

Abstract Background: Tissue engineering technology has been applied extensively for clinical research and human amnion mesenchymal stem cells (hAMSCs) could cause mesenchymal stem cells to differentiate into the bone tissue. However, it is necessary to develop and identify the safer appropriate amount of osteogenic inducer. The objective of this study is to investigate the effect of icariin (ICA) on the proliferation and osteogenic differentiation of hAMSCs.Methods: The morphology and phenotype of hAMSCs were discovered by flow cytometry and immunocytochemical staining. The osteogenic differentiation of hAMSCs under the influence of different concentrations of ICA were assessed by Alkaline phosphatase (ALP) activity substrate assay and Alizarin red Staining.Results: MTT assay revealed that the hAMSCs pretreated with ICA exhibited increased proliferation when compared with the control group, and the most optimum concentration of ICA was 1×10-6 mol/L. The combined analysis of ALP activity and ARS staining showed that ICA could significantly promote the osteogenic differentiation of hAMSCs, and the effect was most significant when the concentration of ICA was 1×10-6mol/L.Conclusion: All above results implied that ICA could significantly increase proliferation and enhanced the osteogenic differentiation of hAMSCs, especially when the concentration of ICA was 1×10-6 mol/L.Keywords: Human amniotic mesenchymal stem cell, Icariin, Proliferation, Osteogenic differentiation

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