In vitro Investigation of Mesenchymal Stem Cells with Nanophase PLGA/HA Composite

Bone grafts have been used to fill bone defects caused by disease or trauma. The amount of autografts is limited and allogenic bone grafts may transmit diseases and cause immune responses. Numerous materials have been proposed and used as scaffolds for bone tissue reconstruction. In this study, we tested nanophase PLGA/HA composite with mesenchymal stem cells in vitro to examine its biological response and cellular activity. The nanophase composite was compared to conventional polystyrene on cytocompatibility by cell attachment, proliferation, alkaline phosphotase activity test and scanning electron microscopy (SEM) analysis. The results demonstrated that human mesenchymal cells showed more cell attachment and higher cell proliferation rate when growing on nanophase PLGA/HA composite than those growing on polystyrene alone. And the composite also promoted MSC cells differentiate to osteoblast cells as compared with control. It was suggested that the combination of bone marrow mesenchymal cells with artificial materials or differentiation factors may enhance bone formation and regeneration, nanophase PLGA/HA composite might therefore be a promising scaffold material for bone tissue substitute in clinical application.

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Hybrid Sponge-Like Scaffolds Based on Ulvan and Gelatin: Design, Characterization and Evaluation of Their Potential Use in Bone Tissue Engineering

Materials ◽

10.3390/ma13071763 ◽

2020 ◽

Vol 13 (7) ◽

pp. 1763

Author(s):

Leto-Aikaterini Tziveleka ◽

Andreas Sapalidis ◽

Stefanos Kikionis ◽

Eleni Aggelidou ◽

Efterpi Demiri ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Mechanical Performance ◽

Cell Attachment ◽

Sulfated Polysaccharide ◽

Hybrid Scaffolds

Ulvan, a bioactive natural sulfated polysaccharide, and gelatin, a collagen-derived biopolymer, have attracted interest for the preparation of biomaterials for different biomedical applications, due to their demonstrated compatibility for cell attachment and proliferation. Both ulvan and gelatin have exhibited osteoinductive potential, either alone or in combination with other materials. In the current work, a series of novel hybrid scaffolds based on crosslinked ulvan and gelatin was designed, prepared and characterized. Their mechanical performance, thermal stability, porosity, water-uptake and in vitro degradation ability were assessed, while their morphology was analyzed through scanning electron microscopy. The prepared hybrid ulvan/gelatin scaffolds were characterized by a highly porous and interconnected structure. Human adipose-derived mesenchymal stem cells (hADMSCs) were seeded in selected ulvan/gelatin hybrid scaffolds and their adhesion, survival, proliferation, and osteogenic differentiation efficiency was evaluated. Overall, it was found that the prepared hybrid sponge-like scaffolds could efficiently support mesenchymal stem cells’ adhesion and proliferation, suggesting that such scaffolds could have potential uses in bone tissue engineering.

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Gelatin/Hydroxyapatite Scaffolds: Studies on Adhesion, Growth and Differentiation of Mesenchymal Stem Cells

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.93-94.121 ◽

2010 ◽

Vol 93-94 ◽

pp. 121-124

Author(s):

Nuttapon Vachiraroj ◽

Siriporn Damrongsakkul ◽

Sorada Kanokpanont

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Calcium Content ◽

Population Doubling ◽

Population Doubling Time ◽

3 Dimensional

In this work, we developed a 3-dimensional bone tissue engineering scaffold from type B gelatin and hydroxyapatite. Two types of scaffolds, pure gelatin (pI~5) (Gel) and gelatin/hydroxyapatite (30/70 wt./wt.) (Gel/HA), were prepared from concentrated solutions (5% wt./wt.) using foaming/freeze drying method. The results SEM revealed the interconnected-homogeneous pores of Gel and Gel/HA were 121  119 and 148  83m, respectively. Hydroxyapatite improved mechanical property of the gelatin scaffolds, especially at dry state. Compressive modulus of Gel and Gel/HA scaffolds were at 118±21.68 and 510±109.08 kPa, respectively. The results on in vitro cells culture showed that Gel/HA scaffolds promoted attachment of rat’s mesenchymal stem cells (MSC) to a 1.23 folds higher than the Gel scaffolds. Population doubling time (PDT) of MSC on Gel and Gel/HA scaffolds were 51.16 and 54.89 hours, respectively. In term of osteogenic differentiation, Gel/HA scaffolds tended to enhance ALP activity and calcium content of MSC better than those of the Gel scaffold. Therefore the Gel/HA scaffolds had a potential to be applied in bone tissue engineering.

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Mesenchymal Stem Cells Exhibit Both a Proinflammatory and Anti-Inflammatory Effect on Saccular Aneurysm Formation in a Rabbit Model

Stem Cells International ◽

10.1155/2019/3618217 ◽

2019 ◽

Vol 2019 ◽

pp. 1-15

Author(s):

Michael B. Avery ◽

Brooke L. Belanger ◽

Amy Bromley ◽

Arindom Sen ◽

Alim P. Mitha

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Rabbit Model ◽

Potential Interaction ◽

Allogenic Bone ◽

Aneurysm Formation ◽

Vehicle Injection ◽

Anti Inflammatory

Several studies have demonstrated a potential interaction between mesenchymal stem cells (MSCs) and saccular aneurysms. In this study, we sought to determine whether allogenic bone marrow-derived MSCs had the ability to prevent aneurysm formation in a known rabbit elastase aneurysm model. MSCs were injected intravenously in experimental rabbits at the time of surgical creation and two weeks postcreation and compared with control rabbits receiving vehicle injection. Angiography was used to compare aneurysm measurements four weeks postcreation, and aneurysms were harvested for histological properties. Serum was collected longitudinally to evaluate cytokine alterations. Serum from control animals was also utilized to perform in vitro tests with MSCs to compare the effect of the serologic environment in animals with and without aneurysms on MSC proliferation and cytokine production. While aneurysm morphometric comparisons revealed no differences, significant cytokine alterations were observed in vitro and in vivo, suggesting both anti-inflammatory and proinflammatory processes were occurring in the presence of MSCs. Histological analyses suggested that tunica intima hyperplasia was inhibited in the presence of MSCs.

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Acceleration of Bone Regeneration in Critical-Size Defect Using BMP-9-Loaded nHA/ColI/MWCNTs Scaffolds Seeded with Bone Marrow Mesenchymal Stem Cells

BioMed Research International ◽

10.1155/2019/7343957 ◽

2019 ◽

Vol 2019 ◽

pp. 1-10 ◽

Cited By ~ 2

Author(s):

Ran Zhang ◽

Xuewen Li ◽

Yao Liu ◽

Xiaobo Gao ◽

Tong Zhu ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Composite Scaffold ◽

Marrow Mesenchymal Stem Cells

Biocompatible scaffolding materials play an important role in bone tissue engineering. This study sought to develop and characterize a nano-hydroxyapatite (nHA)/collagen I (ColI)/multi-walled carbon nanotube (MWCNT) composite scaffold loaded with recombinant bone morphogenetic protein-9 (BMP-9) for bone tissue engineering by in vitro and in vivo experiments. The composite nHA/ColI/MWCNT scaffolds were fabricated at various concentrations of MWCNTs (0.5, 1, and 1.5% wt) by blending and freeze drying. The porosity, swelling rate, water absorption rate, mechanical properties, and biocompatibility of scaffolds were measured. After loading with BMP-9, bone marrow mesenchymal stem cells (BMMSCs) were seeded to evaluate their characteristics in vitro and in a critical sized defect in Sprague-Dawley rats in vivo. It was shown that the 1% MWCNT group was the most suitable for bone tissue engineering. Our results demonstrated that scaffolds loaded with BMP-9 promoted differentiation of BMMSCs into osteoblasts in vitro and induced more bone formation in vivo. To conclude, nHA/ColI/MWCNT scaffolds loaded with BMP-9 possess high biocompatibility and osteogenesis and are a good candidate for use in bone tissue engineering.

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Osteogenic potential of mesenchymal stem cells from rat mandible to regenerate critical sized calvarial defect

Journal of Tissue Engineering ◽

10.1177/2041731419830427 ◽

2019 ◽

Vol 10 ◽

pp. 204173141983042 ◽

Cited By ~ 9

Author(s):

Dong Joon Lee ◽

Jane Kwon ◽

Luke Current ◽

Kun Yoon ◽

Rahim Zalal ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Osteogenic Potential ◽

Embryonic Origin ◽

Rat Mandible

Although bone marrow–derived mesenchymal stem cells (MSCs) have been extensively explored in bone tissue engineering, only few studies using mesenchymal stem cells from mandible (M-MSCs) have been reported. However, mesenchymal stem cells from mandible have the potential to be as effective as femur-derived mesenchymal stem cells (F-MSCs) in regenerating bone, especially in the orofacial regions, which share embryonic origin, proximity, and accessibility. M-MSCs were isolated and characterized using mesenchymal stem cell–specific markers, colony forming assay, and multi-potential differentiation. In vitro osteogenic potential, including proliferation, osteogenic gene expression, alkaline phosphatase activity, and mineralization, was examined and compared. Furthermore, in vivo bone formations of F-MSCs and M-MSCs in rat critical sized defect were evaluated using microCT and histology. M-MSCs from rat could be successfully isolated and expanded while preserving their MSC’s characteristics. M-MSCs demonstrated a comparable proliferation and mineralization potentials and in vivo bone formation as F-MSCs. M-MSCs is a promising cell source candidate for craniofacial bone tissue engineering.

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Intrinsic Cellular Responses of Human Wharton’s Jelly Mesenchymal Stem Cells Influenced by O2-Plasma-Modified and Unmodified Surface of Alkaline-Hydrolyzed 2D and 3D PCL Scaffolds

Journal of Functional Biomaterials ◽

10.3390/jfb10040052 ◽

2019 ◽

Vol 10 (4) ◽

pp. 52 ◽

Cited By ~ 1

Author(s):

Inthanon ◽

Janvikul ◽

Ongchai ◽

Chomdej

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Cell Attachment ◽

Physicochemical Characterization ◽

Wharton’S Jelly ◽

Wharton's Jelly ◽

Differentiation Inducers ◽

2D And 3D

Polycaprolactone (PCL), a hydrophobic-degradable polyester, has been widely investigated and extensively developed, to increase the biocompatibility for tissue engineering. This research was the first trial to evaluate the intrinsic biological responses of human Wharton’s Jelly Mesenchymal Stem Cells (hWJMSCs) cultured on alkaline hydrolysis and low-pressure oxygen plasma modified 2D and 3D PCL scaffolds, without adding any differentiation inducers; this has not been reported before. Four types of the substrate were newly established: 2D plasma-treated PCL (2D-TP), 2D non-plasma-treated PCL (2D-NP), 3D plasma-treated PCL (3D-TP), and 3D non-plasma-treated PCL (3D-NP). Physicochemical characterization revealed that only plasma-treated PCL scaffolds significantly increased the hydrophilicity and % oxygen/carbon ratio on the surfaces. The RMS roughness of 3D was higher than 2D conformation, whilst the plasma-treated surfaces were rougher than the non-plasma treated ones. The cytocompatibility test demonstrated that the 2D PCLs enhanced the initial cell attachment in comparison to the 3Ds, indicated by a higher expression of focal adhesion kinase. Meanwhile, the 3Ds promoted cell proliferation and migration as evidence of higher cyclin-A expression and filopodial protrusion, respectively. The 3Ds potentially protected the cell from apoptosis/necrosis but also altered the pluripotency/differentiation-related gene expression. In summary, the different configuration and surface properties of PCL scaffolds displayed the significant potential and effectiveness for facilitating stem cell growth and differentiation in vitro. The cell–substrate interactions on modified surface PCL may provide some information which could be further applied in substrate architecture for stem cell accommodation in cell delivery system for tissue repair.

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A Novel Strategy Incorporated the Power of Mesenchymal Stem Cells to Allografts for Segmental Bone Tissue Engineering

Cell Transplantation ◽

10.3727/096368909788809839 ◽

2009 ◽

Vol 18 (4) ◽

pp. 433-441 ◽

Cited By ~ 45

Author(s):

Xiao Hui Zou ◽

Hong Xin Cai ◽

Zi Yin ◽

Xiao Chen ◽

Yang Zi Jiang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Regeneration ◽

Nude Mice ◽

Bone Grafts ◽

Great Promise ◽

Differentiation Potential ◽

Bone Allografts ◽

Segmental Bone

Mesenchymal stem cells (MSCs) hold great promise for bone regeneration. However, the power of mesenchymal stem cells has not been applied to structural bone allografts in clinical practice. This study designed a new strategy to enhance the efficiency of allografts for segmental bone regeneration. Isolated MSCs were cultured to form a cell sheet. The MSC sheet was then wrapped onto structural allografts. The assembled structures were cultured in vitro to evaluate the differentiation potential of MSC sheet. The assembled structures were implanted subcutaneously into nude mice as well as into the segmental radius defect of rabbits to investigate the efficiency of MSC sheets to repopulate allografts for bone repair. MSC sheets, upon assembling on bone grafts, showed similar differentiation properties to the in situ periosteum in vitro. After implantation the MSC sheets accelerated the repopulation of bone grafts in nude mice. Moreover, MSC sheets induced thicker cortical bone formation and more efficient graft-to-bone end fusion at the segmental bone defects in rabbits. This study thus presented a novel, more efficient, and practical strategy for large weight-bearing bone reconstruction by using MSC sheets to deliver large number of MSCs to repopulate the bone allografts.

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Increased COUP-TFII Expression Mediates the Differentiation Imbalance of Bone Marrow-Derived Mesenchymal Stem Cells in Femoral Head Osteonecrosis

BioMed Research International ◽

10.1155/2019/9262430 ◽

2019 ◽

Vol 2019 ◽

pp. 1-11 ◽

Cited By ~ 2

Author(s):

Sheng-Hao Wang ◽

Guo-Hau Gou ◽

Chia-Chun Wu ◽

Hsain-Chung Shen ◽

Leou-Chyr Lin ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Femoral Head ◽

Bone Tissue ◽

Mesenchymal Cells ◽

Control Group ◽

Differentiation Potential ◽

Femoral Head Osteonecrosis ◽

Neck Fracture

Objective. Bone marrow-derived mesenchymal stem cells (BMSCs) have multilineage differentiation potential, which allows them to progress to osteogenesis, adipogenesis, and chondrogenesis. An imbalance of differentiation between osteogenesis and adipogenesis will result in pathologic conditions inside the bone. This type of imbalance is also one of the pathological findings in osteonecrosis of the femoral head (ONFH). Chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) was previously reported to mediate the differentiation of mesenchymal stem cells. This study investigated the expression of the osteogenesis regulator Runx2, osteocalcin, the adipogenesis regulator PPARγ, and COUP-TFII in the femoral head tissue harvested from ONFH patients, and characterized the effect of COUP-TFII on the differentiation of primary BMSCs. Methods. Thirty patients with ONFH were recruited and separated into 3 groups: the trauma-, steroid- and alcohol-induced ONFH groups (10 patients each). Bone specimens were harvested from patients who underwent hip arthroplasty, and another 10 specimens were harvested from femoral neck fracture patients as the control group. Expression of the osteogenesis regulator Runx2, osteocalcin, the adipogenesis regulator PPARγ, C/EBP-α, and COUP-TFII was analyzed by Western blotting. Primary bone marrow mesenchymal cells were harvested from ONFH cells treated with COUP-TFII RNA interference to evaluate the effect of COUP-TFII on MSCs. Results. ONFH patients had significantly increased expression of the adipogenesis regulator PPARγ and C/EBP-α and decreased expression of the osteogenesis regulator osteocalcin. ONFH bone tissue also revealed higher COUP-TFII expression. Immunohistochemical staining displayed strong COUP-TFII immunoreactivity adjacent to osteonecrotic trabecular bone. Increased COUP-TFII expression in the bone tissue correlated with increased PPARγ and decreased osteocalcin expression. Knockdown of COUP-TFII with siRNA in BMSCs reduced adipogenesis and increased osteogenesis in mesenchymal cells. Conclusion. Increased COUP-TFII expression mediates the imbalance of BMSC differentiation and progression to ONFH in patients. This study might reveal a new target in the treatment of ONFH.

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Polycaprolactone-Hydroxyapatite Composite Membrane Scaffolds for Bone Tissue Engineering

MRS Proceedings ◽

10.1557/opl.2013.567 ◽

2013 ◽

Vol 1502 ◽

Cited By ~ 3

Author(s):

Sabrina Morelli ◽

Daniele Facciolo ◽

Antonietta Messina ◽

Antonella Piscioneri ◽

Simona Salerno ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Composite Membrane ◽

Cell Attachment ◽

Cellular Systems ◽

Phase Inversion Technique ◽

Membrane Scaffold

ABSTRACTBone tissue engineering typically involves the use of porous, bioresorbable scaffolds to serve as temporary, three-dimensional scaffolds to guide cell attachment, differentiation, proliferation, and subsequent tissue regeneration. In this study we developed a composite membrane scaffold by phase inversion technique by using biodegradable polyester, Polycaprolactone (PCL), with hydroxyapatite (HA) in order to develop novel controlled nanostructured biomaterials for bone tissue engineering applications.After preparation, membrane scaffolds were characterized in order to evaluate its morphological, physico-chemical and mechanical properties and then used for the cell culture.Our experimental design consists to apply the knowledge of natural bone tissue remodelling in an in vitro membrane biohybrid system. We used human mesenchymal stem cells for culture in the membrane scaffolds inducing the differentiation in osteoblasts and human monocytes to trigger osteoclastogenesis. Osteoclastic resorption of the scaffold material would lead to subsequent induction of osteoblasts and faster bone formation with mesenchymal stem cells. Our results show that osteoblasts and osteoclasts were successfully differentiated in the developed PCL-HA membrane scaffold. This membrane system will lead to insights in the creation of a controllable osteoinductive microenvironment based on the specific properties (e.g. basic composition, surface chemistry, architecture) and on the function (resorption coupled to proliferation and differentiation) of defined cellular systems.

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