scholarly journals An Innovative Laboratory Procedure to Expand Chondrocytes with Reduced Dedifferentiation

Cartilage ◽  
2017 ◽  
Vol 9 (2) ◽  
pp. 202-211 ◽  
Author(s):  
Yong Mao ◽  
Tyler Hoffman ◽  
Amy Wu ◽  
Joachim Kohn
Keyword(s):  
Tissue Engineering ◽  
Tissue Culture ◽  
Nude Mice ◽  
Cartilage Tissue Engineering ◽  
Basic Research ◽  
Cartilage Tissue ◽  
Laboratory Procedure ◽  
In Vitro Expansion ◽  
Culture Surface

Objective In vitro expansion of chondrocytes is required for cartilage tissue engineering and clinical cell-based cartilage repair practices. However, the dedifferentiation of chondrocytes during in vitro expansion continues to be a challenge. This study focuses on identifying a cell culture surface to support chondrocyte expansion with reduced dedifferentiation. Design A less adhesive culture surface, non–tissue culture treated surface (NTC), was tested for its suitability for culturing chondrocytes. The cell expansion and the expression of chondrocyte markers were monitored for at least 2 passages on NTC in comparison with conventional tissue culture treated polystyrene surface (TCP). The ability of expanded chondrocytes to form cartilage tissues was evaluated using pellet culturing and subcutaneous implantation in nude mice. Results NTC supported bovine chondrocyte proliferation to a clinically relevant expansion requirement within 2 passages. Chondrocyte phenotypes were better maintained when cultured on NTC than on TCP. In vitro pellet culture studies showed that chondrocytes expanded on NTC expressed a higher level of chondrocyte extracellular matrix. Furthermore, the cells expanded on NTC or TCP were implanted subcutaneously as pellets in nude mice for 6 weeks. The recovered pellets showed cartilage-like tissue formation from cells expanded on NTC but not from the cells expanded on TCP. Conclusions This study presents an innovative and easy culturing procedure to expand chondrocytes with reduced dedifferentiation. This procedure has potential to be developed to expand chondrocytes in vitro for basic research, tissue engineering, and possibly for clinical applications.

scholarly journals Neural Crest-Derived Chondrocytes Isolation for Tissue Engineering in Regenerative Medicine

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 962
Author(s):  
Monica Salamone ◽  
Salvatrice Rigogliuso ◽  
Aldo Nicosia ◽  
Marcello Tagliavia ◽  
Simona Campora ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cartilage Tissue Engineering ◽  
Enzymatic Digestion ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Nasal Cartilage ◽  
Tissue Dissociation ◽  
In Vitro Expansion ◽  
First Time

Chondrocyte transplantation has been successfully tested and proposed as a clinical procedure aiming to repair articular cartilage defects. However, the isolation of chondrocytes and the optimization of the enzymatic digestion process, as well as their successful in vitro expansion, remain the main challenges in cartilage tissue engineering. In order to address these issues, we investigated the performance of recombinant collagenases in tissue dissociation assays with the aim of isolating chondrocytes from bovine nasal cartilage in order to establish the optimal enzyme blend to ensure the best outcomes of the overall procedure. We show, for the first time, that collagenase H activity alone is required for effective cartilage digestion, resulting in an improvement in the yield of viable cells. The extracted chondrocytes proved able to grow and activate differentiation/dedifferentiation programs, as assessed by morphological and gene expression analyses.

Platelet lysate reduces the chondrocyte dedifferentiation during in vitro expansion: Implications for cartilage tissue engineering

2020 ◽  
Vol 133 ◽  
pp. 98-105 ◽  
Author(s):  
Elena De Angelis ◽  
Stefano Grolli ◽  
Roberta Saleri ◽  
Virna Conti ◽  
Melania Andrani ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cartilage Tissue Engineering ◽  
Platelet Lysate ◽  
Cartilage Tissue ◽  
In Vitro Expansion

scholarly journals Potential of 3-D tissue constructs engineered from bovine chondrocytes/silk fibroin-chitosan for in vitro cartilage tissue engineering

Biomaterials ◽  
2011 ◽  
Vol 32 (25) ◽  
pp. 5773-5781 ◽  
Author(s):  
Nandana Bhardwaj ◽  
Quynhhoa T. Nguyen ◽  
Albert C. Chen ◽  
David L. Kaplan ◽  
Robert L. Sah ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Silk Fibroin ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Tissue Constructs

scholarly journals Calcium/Cobalt Alginate Beads as Functional Scaffolds for Cartilage Tissue Engineering

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Stefano Focaroli ◽  
Gabriella Teti ◽  
Viviana Salvatore ◽  
Isabella Orienti ◽  
Mirella Falconi
Keyword(s):  
Tissue Engineering ◽  
Bone Morphogenetic Proteins ◽  
Transforming Growth Factor ◽  
Cartilage Tissue Engineering ◽  
Biomechanical Properties ◽  
Alginate Beads ◽  
Cartilage Tissue ◽  
Self Healing ◽  
Tissue Replacement

Articular cartilage is a highly organized tissue with complex biomechanical properties. However, injuries to the cartilage usually lead to numerous health concerns and often culminate in disabling symptoms, due to the poor intrinsic capacity of this tissue for self-healing. Although various approaches are proposed for the regeneration of cartilage, its repair still represents an enormous challenge for orthopedic surgeons. The field of tissue engineering currently offers some of the most promising strategies for cartilage restoration, in which assorted biomaterials and cell-based therapies are combined to develop new therapeutic regimens for tissue replacement. The current study describes thein vitrobehavior of human adipose-derived mesenchymal stem cells (hADSCs) encapsulated within calcium/cobalt (Ca/Co) alginate beads. These novel chondrogenesis-promoting scaffolds take advantage of the synergy between the alginate matrix and Co+2ions, without employing costly growth factors (e.g., transforming growth factor betas (TGF-βs) or bone morphogenetic proteins (BMPs)) to direct hADSC differentiation into cartilage-producing chondrocytes.

TRENDS AND CHALLENGES OF CARTILAGE TISSUE ENGINEERING

2009 ◽  
Vol 21 (03) ◽  
pp. 149-155 ◽  
Author(s):  
Hsu-Wei Fang
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Growth Factors ◽  
Bone Cells ◽  
Cartilage Tissue Engineering ◽  
Bioactive Molecules ◽  
In Vivo Studies ◽  
Cartilage Tissue

Cartilage injuries may be caused by trauma, biomechanical imbalance, or degenerative changes of joint. Unfortunately, cartilage has limited capability to spontaneous repair once damaged and may lead to progressive damage and degeneration. Cartilage tissue-engineering techniques have emerged as the potential clinical strategies. An ideal tissue-engineering approach to cartilage repair should offer good integration into both the host cartilage and the subchondral bone. Cells, scaffolds, and growth factors make up the tissue engineering triad. One of the major challenges for cartilage tissue engineering is cell source and cell numbers. Due to the limitations of proliferation for mature chondrocytes, current studies have alternated to use stem cells as a potential source. In the recent years, a lot of novel biomaterials has been continuously developed and investigated in various in vitro and in vivo studies for cartilage tissue engineering. Moreover, stimulatory factors such as bioactive molecules have been explored to induce or enhance cartilage formation. Growth factors and other additives could be added into culture media in vitro, transferred into cells, or incorporated into scaffolds for in vivo delivery to promote cellular differentiation and tissue regeneration.Based on the current development of cartilage tissue engineering, there exist challenges to overcome. How to manipulate the interactions between cells, scaffold, and signals to achieve the moderation of implanted composite differentiate into moderate stem cells to differentiate into hyaline cartilage to perform the optimum physiological and biomechanical functions without negative side effects remains the target to pursue.

scholarly journals In vitro characterization of a novel magnetic fibrin-agarose hydrogel for cartilage tissue engineering

2020 ◽  
Vol 104 ◽  
pp. 103619 ◽  
Author(s):  
Ana Belén Bonhome-Espinosa ◽  
Fernando Campos ◽  
Daniel Durand-Herrera ◽  
José Darío Sánchez-López ◽  
Sébastien Schaub ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
In Vitro Characterization ◽  
Agarose Hydrogel

Poly-N-Acetyl Glucosamine as a Scaffold for Cartilage Tissue Engineering in Nude Mice

2005 ◽  
pp. 71-74
Author(s):  
Christopher M. Hill ◽  
Yuehuei H. An ◽  
Qian K. Kang ◽  
Marina V. Demcheva ◽  
S.William Whitson ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Nude Mice ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue

scholarly journals 23.4 The combination of dynamic compression and shear with rvBMP-2 for in-vitro cartilage tissue engineering

2007 ◽  
Vol 15 ◽  
pp. B81
Author(s):  
G.M. Salzmann ◽  
P. Schmitz ◽  
M. Anton ◽  
M. Stoddart ◽  
S. Grad ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Dynamic Compression ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue

Mechano-Active Cartilage Tissue Engineering

2006 ◽  
Vol 49 ◽  
pp. 189-196
Author(s):  
Soo Hyun Kim ◽  
Young Mee Jung ◽  
Sang Heon Kim ◽  
Young Ha Kim ◽  
Jun Xie ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Dynamic Compression ◽  
Cartilage Tissue Engineering ◽  
Compressive Deformation ◽  
Cartilage Tissue ◽  
Cartilaginous Tissue ◽  
Pressing Method ◽  
Safranin O

To engineer cartilaginous constructs with a mechano-active scaffold and dynamic compression was performed for effective cartilage tissue engineering. Mechano-active scaffolds were fabricated from very elastic poly(L-lactide-co-ε-carprolactone)(5:5). The scaffolds with 85 % porosity and 300~500 μm pore size were prepared by a gel-pressing method. The scaffolds were seeded with chondrocytes and the continuous compressive deformation of 5% strain was applied to cell-polymer constructs with 0.1Hz to evaluate for the effect of dynamic compression for regeneration of cartilage. Also, the chondrocytes-seeded constructs stimulated by the continuous compressive deformation of 5% strain with 0.1Hz for 10 days and 24 days respectively were implanted in nude mice subcutaneously to investigate their biocompatibility and cartilage formation. From biochemical analyses, chondrogenic differentiation was sustained and enhanced significantly and chondrial extracellular matrix was increased through mechanical stimulation. Histological analysis showed that implants stimulated mechanically formed mature and well-developed cartilaginous tissue, as evidenced by chondrocytes within lacunae. Masson’s trichrome and Safranin O staining indicated an abundant accumulation of collagens and GAGs. Also, ECM in constructs was strongly immuno-stained with anti-rabbit collagen type II antibody. Consequently, the periodic application of dynamic compression can improve the quality of cartilaginous tissue formed in vitro and in vivo.

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