scholarly journals Biomimetic Cell-Laden MeHA Hydrogels for the Regeneration of Cartilage Tissue

Polymers ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 1598
Author(s):  
Evgenia Tsanaktsidou ◽  
Olga Kammona ◽  
Norina Labude ◽  
Sabine Neuss ◽  
Melanie Krüger ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Human Mesenchymal Stem Cells ◽  
Cartilage Tissue ◽  
Induction Medium ◽  
Encapsulated Cells ◽  
Tissue Culture Polystyrene ◽  
Matrix Metalloproteinase 7 ◽  
Chondrogenic Phenotype ◽  
Polystyrene Plate ◽  
Stem Cell Medium

Methacrylated hyaluronic acid (MeHA) and chondroitin sulfate (CS)-biofunctionalized MeHA (CS-MeHA), were crosslinked in the presence of a matrix metalloproteinase 7 (MMP7)-sensitive peptide. The synthesized hydrogels were embedded with either human mesenchymal stem cells (hMSCs) or chondrocytes, at low concentrations, and subsequently cultured in a stem cell medium (SCM) or chondrogenic induction medium (CiM). The pivotal role of the synthesized hydrogels in promoting the expression of cartilage-related genes and the formation of neocartilage tissue despite the low concentration of encapsulated cells was assessed. It was found that hMSC-laden MeHA hydrogels cultured in an expansion medium exhibited a significant increase in the expression of chondrogenic markers compared to hMSCs cultured on a tissue culture polystyrene plate (TCPS). This favorable outcome was further enhanced for hMSC-laden CS-MeHA hydrogels, indicating the positive effect of the glycosaminoglycan binding peptide on the differentiation of hMSCs towards a chondrogenic phenotype. However, it was shown that an induction medium is necessary to achieve full span chondrogenesis. Finally, the histological analysis of chondrocyte-laden MeHA hydrogels cultured on an ex vivo osteochondral platform revealed the deposition of glycosaminoglycans (GAGs) and the arrangement of chondrocyte clusters in isogenous groups, which is characteristic of hyaline cartilage morphology.

scholarly journals Generation and Evaluation of Novel Biomaterials Based on Decellularized Sturgeon Cartilage for Use in Tissue Engineering

Biomedicines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 775
Author(s):  
Olimpia Ortiz-Arrabal ◽  
Ramón Carmona ◽  
Óscar-Darío García-García ◽  
Jesús Chato-Astrain ◽  
David Sánchez-Porras ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Ex Vivo ◽  
Cartilage Damage ◽  
Human Mesenchymal Stem Cells ◽  
In Silico Analysis ◽  
Cartilage Tissue ◽  
Advanced Therapy Medicinal Product ◽  
Advanced Therapy ◽  
Novel Scaffold

Because cartilage has limited regenerative capability, a fully efficient advanced therapy medicinal product is needed to treat severe cartilage damage. We evaluated a novel biomaterial obtained by decellularizing sturgeon chondral endoskeleton tissue for use in cartilage tissue engineering. In silico analysis suggested high homology between human and sturgeon collagen proteins, and ultra-performance liquid chromatography confirmed that both types of cartilage consisted mainly of the same amino acids. Decellularized sturgeon cartilage was recellularized with human chondrocytes and four types of human mesenchymal stem cells (MSC) and their suitability for generating a cartilage substitute was assessed ex vivo and in vivo. The results supported the biocompatibility of the novel scaffold, as well as its ability to sustain cell adhesion, proliferation and differentiation. In vivo assays showed that the MSC cells in grafted cartilage disks were biosynthetically active and able to remodel the extracellular matrix of cartilage substitutes, with the production of type II collagen and other relevant components, especially when adipose tissue MSC were used. In addition, these cartilage substitutes triggered a pro-regenerative reaction mediated by CD206-positive M2 macrophages. These preliminary results warrant further research to characterize in greater detail the potential clinical translation of these novel cartilage substitutes.

Transient Exposure to TGF-β3 Improves the Functional Properties of MSC-Seeded Photocrosslinked Hyaluronic Acid Hydrogels

2011 ◽  
Author(s):  
Minwook Kim ◽  
Isaac E. Erickson ◽  
Jason A. Burdick ◽  
Robert L. Mauck
Keyword(s):  
Hyaluronic Acid ◽  
Functional Properties ◽  
Cartilage Tissue Engineering ◽  
Compressive Load ◽  
Cartilage Tissue ◽  
Attractive Alternative ◽  
Biologically Relevant ◽  
Encapsulated Cells ◽  
Chondrogenic Phenotype

Articular cartilage is the primary compressive load bearing soft tissue in diarthrodial joints. While the tissue can function remarkably well in a demanding environment over a lifetime of use, focal defects and other trauma can initiate progressive degeneration. Cartilage tissue engineering approaches have been developed with the goal of forming biologic replacement materials with functional mechanical properties [1]. While chondrocytes are a popular cell source for such approaches, and can produce constructs with near-native functional properties [2], mesenchymal stem cells (MSCs) derived from bone marrow have emerged as an attractive alternative cell type. MSCs are multi-potent and easy to expand, and so are available in a nearly unlimited supply, and in an autologous fashion. While MSCs can undergo functional chondrogenesis in a variety of 3D contexts [3], we are particularly interested in the translational capacity of hyaluronic acid (HA). Hydrogels formed from this natural constituent of the cartilage extracellular matrix provide a biologically relevant interface for encapsulated cells and gel properties are readily tunable [4, 5]. Indeed, using a methacrylated (and so photo-crosslinkable) HA macromer, we have optimized gel formation and functional matrix production by MSCs with variations in both macromer (1%, [5]) and MSC (∼60 million cells/mL, [6]) concentration, consistently producing cartilage-like constructs with near native compressive properties. Additionally, we have reported that transient exposure of TGF-β3 (for three weeks) to MSCs in agarose constructs at a high-density induced a stable chondrogenic phenotype, with functional properties at six weeks greater than continual exposure to this pro-chondrogenic factor [7]. Transient exposure presents an interesting paradigm with clinical relevance, in vivo defect filling will require robust maturation of the engineered tissue driven by TGF-β3 delivered from the material itself in a controlled and sustained fashion. The purpose of this study was to determine the minimal TGF-β3 dosage and duration of exposure required to promote the most robust chondrogenesis and functional maturation of MSCs in this HA hydrogel system.

scholarly journals 3D soft hydrogels induce human mesenchymal stem cells “deep” quiescence

2021 ◽  
Author(s):  
David Boaventura Gomes ◽  
Ana Filipa Henriques Lourenço ◽  
Clarissa Tomasina ◽  
Bryan Chömpff ◽  
Hong Liu ◽  
...  
Keyword(s):  
Kinase Inhibitor ◽  
Ex Vivo ◽  
Mammalian Target Of Rapamycin ◽  
Human Mesenchymal Stem Cells ◽  
Quiescent State ◽  
Mechanical Stiffness ◽  
Great Opportunity ◽  
Cell Therapies ◽  
Tissue Culture Polystyrene ◽  
Protein Levels

AbstractHuman mesenchymal stem/stromal cells (hMSCs) present a great opportunity for tissue regeneration due to their multipotent capacity. However, when cultured on 2D tissue culture polystyrene (TCPS) plates, hMSCs lose their differentiation capacity and clinical potential. It has been reported that cells need a more physiologically relevant micro-environment that allows them to maintain their phenotype. Here, we have developed a 3D alginate hydrogel functionalized with the Arg-Gly-Asp (RGD) sequence and having low mechanical stiffness that mimics the mechanical properties (>5 KPa) of bone marrow. hMSCs cultured in these hydrogels appeared to be halted in G1 phase of the cell cycle and to be non-proliferative, as shown by flow cytometry and 5-Ethynyl-2’-deoxyuridine (EdU) staining, respectively. Their quiescent state was characterized by an upregulation of enhancer of zeste homolog 1 (EZH1) at the gene level, forkhead box O3 (FoxO3) and cyclin-dependent kinase inhibitor 1B (p27) at the gene and protein levels compared to hMSCs grown in 2D TCPS. Comparative studies in 3D hydrogels of alginate-RGD presenting higher concentration of the peptide or in collagen hydrogels revealed that independently of the concentration of RGD or the chemistry of the adhesion motives, hMSCs cultured in 3D presented a similar phenotype.This quiescent phenotype was exclusive of 3D cultures. In 2D, even when cells were starved of fetal bovine serum (FBS) and became also non-proliferative, the expression of these markers was not observed. We propose that this difference may be the result of mammalian target of rapamycin complex 1 (mTORC1) being downregulated in hMSCs cultured in 3D hydrogels, which induces cells to be in “deep” quiescence and be kept alive ex vivo for a long period of time. Our results represent a step forward towards understanding hMSCs quiescence and its molecular pathways, providing more insight for hMSCs cell therapies.

scholarly journals In Vivo Study for Clinical Application of Dental Stem Cell Therapy Incorporated with Dental Titanium Implants

Materials ◽  
2021 ◽  
Vol 14 (2) ◽  
pp. 381
Author(s):  
Hyunmin Choi ◽  
Kyu-Hyung Park ◽  
Narae Jung ◽  
June-Sung Shim ◽  
Hong-Seok Moon ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Formation ◽  
Alveolar Bone ◽  
Human Mesenchymal Stem Cells ◽  
Titanium Implants ◽  
Osteogenic Potential ◽  
Induction Medium ◽  
Radiographic Analysis ◽  
New Bone Formation

The aim of this study was to investigate the behavior of dental-derived human mesenchymal stem cells (d-hMSCs) in response to differently surface-treated implants and to evaluate the effect of d-hMSCs on local osteogenesis around an implant in vivo. d-hMSCs derived from alveolar bone were established and cultured on machined, sandblasted and acid-etched (SLA)-treated titanium discs with and without osteogenic induction medium. Their morphological and osteogenic potential was assessed by scanning electron microscopy (SEM) and real-time polymerase chain reaction (RT-PCR) via mixing of 5 × 106 of d-hMSCs with 1 mL of Metrigel and 20 μL of gel-cell mixture, which was dispensed into the defect followed by the placement of customized mini-implants (machined, SLA-treated implants) in New Zealand white rabbits. Following healing periods of 2 weeks and 12 weeks, the obtained samples in each group were analyzed radiographically, histomorphometrically and immunohistochemically. The quantitative change in osteogenic differentiation of d-hMSCs was identified according to the type of surface treatment. Radiographic analysis revealed that an increase in new bone formation was statistically significant in the d-hMSCs group. Histomorphometric analysis was in accordance with radiographic analysis, showing the significantly increased new bone formation in the d-hMSCs group regardless of time of sacrifice. Human nuclei A was identified near the area where d-hMSCs were implanted but the level of expression was found to be decreased as time passed. Within the limitations of the present study, in this animal model, the transplantation of d-hMSCs enhanced the new bone formation around an implant and the survival and function of the stem cells was experimentally proven up to 12 weeks post-sacrifice.

scholarly journals Allogeneic human mesenchymal stem cells for treatment of E. Coli endotoxin‐induced acute lung injury in an ex vivo perfused human lung

2009 ◽  
Vol 23 (S1) ◽  
Author(s):  
Jae Woo Lee ◽  
Naveen Gupta ◽  
Xiaohui Fang ◽  
James A. Frank ◽  
Raphael Briot ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Acute Lung Injury ◽  
Lung Injury ◽  
Human Lung ◽  
Ex Vivo ◽  
Human Mesenchymal Stem Cells ◽  
E Coli

Food-derived phytochemicals ‘Curcumin’ and ‘Resveratrol’ counteract the catabolic effects of inflammatory lipopolysaccharides: studies on in vitro models of canine and porcine arthritis

2007 ◽  
Vol 2007 ◽  
pp. 8-8
Author(s):  
S.M. Williamson ◽  
J.A. Reynolds ◽  
A. Mobasheri ◽  
M.D. Royal ◽  
A. Vaughan-Thomas ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Structural Damage ◽  
Ex Vivo ◽  
Curcuma Longa ◽  
Companion Animals ◽  
Farm Animals ◽  
Cartilage Tissue ◽  
Feed Conversion ◽  
Growth Plates ◽  
Age Related

Osteoarthritis (OA) and Osteochondritis Dissecans (OCD) are osteoarticular disorders that cause leg weakness, lameness, pain and suffering in companion animals, some farm animals and humans. OA is one of the most common age-related osteoarticular disorders in humans and dogs. In pigs, both OA and OCD are thought to arise from changes in the articular cartilage and growth plates within the synovial joints causing structural damage to joint tissues. Since these changes are not observed in the slow maturing wild boar, they are suggested to be a result of the modern intensive pig production industry which has very successfully selected pigs for rapid growth rates, large muscle mass and efficient feed conversion placing increased weight and mechanical stress on growth plates. The aim of this study was to establish canine and porcine articular cartilage explant models which are essentially tissue culture techniques for isolating and maintaining cartilage tissue ex vivo for subsequent assessment of potentially beneficial effects of specific phytonutrients. Bacterial lipopolysaccharide (LPS) was used as a catabolic mediator to create a culture model of joint inflammation mimicking the events that occur in late stages of OA and OCD. We then performed assays to determine if the dietary phytochemical ‘curcumin’ (derived from Curcuma longa) and the polyphenolic phytoalexin stilbene ‘resveratrol’ (found in red grapes, red wine, peanuts and some berries) are able to counteract the catabolic effects of LPS by inhibiting LPS stimulated release of cartilage matrix glycosaminoglycans (GAGs).

Ex vivo regeneration of the murine kidney from human mesenchymal stem cells

2005 ◽  
Vol 68 (5) ◽  
pp. 1967-1967
Author(s):  
Takashi Yokoo ◽  
Tetsuya Kawamura
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Ex Vivo ◽  
Human Mesenchymal Stem Cells

scholarly journals In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells

2020 ◽  
Vol 21 (2) ◽  
pp. 581 ◽  
Author(s):  
Phoonsuk Limraksasin ◽  
Takeru Kondo ◽  
Maolin Zhang ◽  
Hiroko Okawa ◽  
Thanaphum Osathanon ◽  
...  
Keyword(s):  
Cartilage Tissue ◽  
Marker Genes ◽  
Induction Medium ◽  
Mechanical Stimuli ◽  
Tissue Formation ◽  
Osteogenic Induction Medium ◽  
Osteogenic Induction ◽  
Chondrogenic Marker ◽  
Induced Pluripotent

Cell condensation and mechanical stimuli play roles in osteogenesis and chondrogenesis; thus, they are promising for facilitating self-organizing bone/cartilage tissue formation in vitro from induced pluripotent stem cells (iPSCs). Here, single mouse iPSCs were first seeded in micro-space culture plates to form 3-dimensional spheres. At day 12, iPSC spheres were subjected to shaking culture and maintained in osteogenic induction medium for 31 days (Os induction). In another condition, the osteogenic induction medium was replaced by chondrogenic induction medium at day 22 and maintained for a further 21 days (Os-Chon induction). Os induction produced robust mineralization and some cartilage-like tissue, which promoted expression of osteogenic and chondrogenic marker genes. In contrast, Os-Chon induction resulted in partial mineralization and a large area of cartilage tissue, with greatly increased expression of chondrogenic marker genes along with osterix and collagen 1a1. Os-Chon induction enhanced mesodermal lineage commitment with brachyury expression followed by high expression of lateral plate and paraxial mesoderm marker genes. These results suggest that combined use of micro-space culture and mechanical stimuli facilitates hybrid bone/cartilage tissue formation from iPSCs, and that the bone/cartilage tissue ratio in iPSC constructs could be manipulated through the induction protocol.

scholarly journals Evidence of necroptosis in osteoarthritic disease: investigation of blunt mechanical impact as possible trigger in regulated necrosis

2019 ◽  
Vol 10 (10) ◽  
Author(s):  
Jana Riegger ◽  
Rolf E. Brenner
Keyword(s):  
Cell Death ◽  
Ex Vivo ◽  
Cartilage Explants ◽  
Cartilage Tissue ◽  
Chondrocyte Death ◽  
Intact Cartilage ◽  
Chemically Induced ◽  
Regulated Necrosis ◽  
Enhanced Expression

Abstract Joint injuries are highly associated with cell death and development of posttraumatic osteoarthritis (PTOA). The present study focused on necroptosis as a possible modality of chondrocyte death after cartilage trauma and its relevance in OA disease in general. For this purpose, apoptosis- and necroptosis-associated markers were determined in highly degenerated (ICRS ≥ 3) as well as macroscopically intact cartilage tissue (ICRS ≤ 1) by means of real-time PCR and immunohistochemistry (IHC). Moreover, influence of blunt trauma and/or stimulation with cycloheximide (CHX), TNF-a, and caspase-inhibitor zVAD were investigated in cartilage explants (ICRS ≤ 1). Further characterization of necroptosis was performed in isolated chondrocytes. We found that gene expression levels of RIPK3 (4.2-fold, P < 0.0001) and MLKL (2.7-fold, P < 0.0001) were elevated in highly degenerated cartilage tissue, which was confirmed by IHC staining. After ex vivo trauma and/or CHX/TNF stimulation, addition of zVAD further enhanced expression of necroptosis-related markers as well as release of PGE2 and nitric oxide, which was in line with increased cell death and subsequent release of intracellular HMGB1 and dsDNA in CHX/TNF stimulated chondrocytes. However, trauma and/or chemically induced cell death and subsequent release of pro-inflammatory mediators could be largely attenuated by RIPK1-inhibitor necrostatin 1 or antioxidant N-acetylcysteine. Overall, the study provided clear evidence of necroptotic cell death in OA disease. Moreover, a possible link between cartilage injury and necroptotic processes was found, depending on oxidative stress and cytokine release. These results contribute to further understanding of cell death in PTOA and development of novel therapeutic approaches.

Developmentally Inspired Approach to Cartilage Tissue Engineering

2016 ◽  
Vol 102 ◽  
pp. 31-36
Author(s):  
Esmaiel Jabbari
Keyword(s):  
Articular Cartilage ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Human Mscs ◽  
Superficial Zone ◽  
Developmental Approach ◽  
Encapsulated Cells ◽  
Engineered Tissue ◽  
Zonal Organization ◽  
Articulating Surface

Structural organization of articular cartilage is rooted in the arrangement of mesenchymal stem cells (MSCs) into morphologically distinct zones during embryogenesis as a result of spatiotemporal gradients in biochemical, mechanical, and cellular factors that direct the formation of stratified structure of articular cartilage. These gradients are central to the function of cartilage as an articulating surface. Strategies that mimic zonal organization of articular cartilage are more likely to create an engineered tissue with more effective clinical outcome. The objective of this work was to measure the expression of human MSCs encapsulated in engineered gels that simulate stiffness of the superficial, middle and calcified zones of articular cartilage supplemented with zone specific growth factors. Size of the encapsulated cells increased from the gel simulating superficial zone to those simulating middle and calcified zones. Glycosaminoglycans (GAG) content progressively increased from the gel simulating superficial zone to those simulating middle and calcified zones. Human MSCs in the gel simulating the superficial zone showed up-regulation of Sox-9 and SZP whereas those in the calcified gel showed up-regulation of ALP. Results demonstrate that a developmental approach can potentially regenerate the zonal structure of articular cartilage.

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