scholarly journals Interstitial flow upregulates matrix synthesis and attenuates NF-kB signaling in a novel perfusion bioreactor for articular cartilage tissue engineering

2021 ◽  
Author(s):  
Haneen Abusharkh ◽  
Terreill Robertson ◽  
Juana Mendenhall ◽  
Bulent Gozen ◽  
Edwin Tingstad ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Cell Viability ◽  
Fold Increase ◽  
Cartilage Tissue ◽  
Type I ◽  
Interstitial Flow ◽  
Perfusion System ◽  
Matrix Synthesis ◽  
Porous Chitosan

The present study is focused on designing an easy-to-use novel perfusion system for articular cartilage (AC) tissue engineering and using it to elucidate the mechanism by which interstitial shear upregulates matrix synthesis by articular chondrocytes (AChs). Porous chitosan-agarose (CHAG) scaffolds were synthesized, freeze-dried, and compared to bulk agarose (AG) scaffolds. Both scaffold types were seeded with osteoarthritic human AChs and cultured in a novel perfusion system for one week with a shear-inducing medium flow velocity of 0.33 mm/s corresponding to an average surficial shear of 0.4 mPa and a CHAG interstitial shear of 40 mPa. While there were no statistical differences in cell viability for perfusion vs. static cultures for either scaffold type, CHAG scaffold cultures exhibited 3.3-fold higher (p<0.005) cell viability compared to AG scaffold cultures. Effects of combined superficial and interstitial perfusion for CHAG showed 150- and 45-fold (p<0.0001) increases in total collagen (COL) and 13- and 2.2-fold (p<0.001) increases in glycosaminoglycans (GAGs) over AG’s scaffold non-perfusion and perfusion cultures, respectively, and a 1.5-fold and 3.6-fold (p<0.005) increase over non-perfusion CHAG cultures. Contrasting CHAG perfusion and static cultures, chondrogenic gene comparisons showed a 3.5-fold increase in collagen type II/type I (COL2A1/COL1A1) mRNA ratio (p<0.05), and a 1.3-fold increase in aggrecan mRNA. Observed effects are suggested to be the result of inhibiting the inflammatory NF-κB signal transduction pathway as confirmed by a further study that indicated a reduction by 3.2-fold (p<0.05) upon exposure to perfusion. Our results demonstrate that the presence of pores plays a critical role in improving cell viability and that interstitial flow caused by medium perfusion through the porous scaffolds enhances the expression of chondrogenic genes and ECM components through the downregulation of NF-κB1.

scholarly journals Adipose-Derived Mesenchymal Stem Cell Chondrospheroids Cultured in Hypoxia and a 3D Porous Chitosan/Chitin Nanocrystal Scaffold as a Platform for Cartilage Tissue Engineering

2020 ◽  
Vol 21 (3) ◽  
pp. 1004 ◽  
Author(s):  
Veronica Zubillaga ◽  
Ana Alonso-Varona ◽  
Susana C. M. Fernandes ◽  
Asier M. Salaberria ◽  
Teodoro Palomares
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Collagen Type ◽  
Porous Scaffold ◽  
Cartilage Tissue Engineering ◽  
Cartilage Regeneration ◽  
Cartilage Tissue ◽  
Type I ◽  
Low Oxygen ◽  
Porous Chitosan

Articular cartilage degeneration is one of the most common causes of pain and disability in middle-aged and older people. Tissue engineering (TE) has shown great therapeutic promise for this condition. The design of cartilage regeneration constructs must take into account the specific characteristics of the cartilaginous matrix, as well as the avascular nature of cartilage and its cells’ peculiar arrangement in isogenic groups. Keeping these factors in mind, we have designed a 3D porous scaffold based on genipin-crosslinked chitosan/chitin nanocrystals for spheroid chondral differentiation of human adipose tissue-derived mesenchymal stem cells (hASCs) induced in hypoxic conditions. First, we demonstrated that, under low oxygen conditions, the chondrospheroids obtained express cartilage-specific markers including collagen type II (COL2A1) and aggrecan, lacking expression of osteogenic differentiation marker collagen type I (COL1A2). These results were associated with an increased expression of hypoxia-inducible factor 1α, which positively directs COL2A1 and aggrecan expression. Finally, we determined the most suitable chondrogenic differentiation pattern when hASC spheroids were seeded in the 3D porous scaffold under hypoxia and obtained a chondral extracellular matrix with a high sulphated glycosaminoglycan content, which is characteristic of articular cartilage. These findings highlight the potential use of such templates in cartilage tissue engineering.

scholarly journals Characterization of Collagen Type I and II Blended Hydrogels for Articular Cartilage Tissue Engineering

2016 ◽  
Vol 17 (10) ◽  
pp. 3145-3152 ◽  
Author(s):  
Nelda Vázquez-Portalatı́n ◽  
Claire E. Kilmer ◽  
Alyssa Panitch ◽  
Julie C. Liu
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Collagen Type ◽  
Cartilage Tissue Engineering ◽  
Collagen Type I ◽  
Cartilage Tissue ◽  
Type I

Development of a Multi-Arm Bioprinter for Hybrid Tissue Engineering

2013 ◽  
Author(s):  
Howard Chen ◽  
Ibrahim T. Ozbolat
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Cell Viability ◽  
Progenitor Cells ◽  
Layer Structure ◽  
Single Layer ◽  
Cartilage Tissue ◽  
High Cell ◽  
Cell Spheroids ◽  
Multiple Materials

This paper highlights the development of a multi-arm bioprinter (MABP) capable of concurrent deposition of multiple materials with independent dispensing parameters including deposition speed, material dispensing rate and frequency for functional zonal-stratified articular cartilage tissue fabrication. The MABP consists of two Cartesian robots mounted in parallel on the same mechanical frame. This platform is used for concurrent filament fabrication and cell spheroid deposition. A single-layer structure is fabricated and concurrently deposited with spheroids to validate this system. Preliminary results showed that the MABP was able to produce filaments and spheroids with well-defined geometry and high cell viability. The resulting filament width has a variation of +/-170 μm and the center-to-center filament distance was within 100 μm of the specified distance. This fabrication system is aimed to be further refined for printing structures with varying porosities to mimic the natural cartilage structure in order to produce functional tissue-engineered articular cartilage using cell spheroids containing cartilage progenitor cells (CPCs).

scholarly journals In vivo cartilage regeneration in a multi-layered articular cartilage architecture mimicking scaffold

2020 ◽  
Vol 9 (9) ◽  
pp. 601-612
Author(s):  
Karthikeyan Rajagopal ◽  
Sowmya Ramesh ◽  
Noel Malcolm Walter ◽  
Aditya Arora ◽  
Dhirendra S. Katti ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cell Viability ◽  
Cartilage Regeneration ◽  
Fold Increase ◽  
Vital Role ◽  
Matrix Synthesis ◽  
Cartilage Formation ◽  
And Function

Aims Extracellular matrix (ECM) and its architecture have a vital role in articular cartilage (AC) structure and function. We hypothesized that a multi-layered chitosan-gelatin (CG) scaffold that resembles ECM, as well as native collagen architecture of AC, will achieve superior chondrogenesis and AC regeneration. We also compared its in vitro and in vivo outcomes with randomly aligned CG scaffold. Methods Rabbit bone marrow mesenchymal stem cells (MSCs) were differentiated into the chondrogenic lineage on scaffolds. Quality of in vitro regenerated cartilage was assessed by cell viability, growth, matrix synthesis, and differentiation. Bilateral osteochondral defects were created in 15 four-month-old male New Zealand white rabbits and segregated into three treatment groups with five in each. The groups were: 1) untreated and allogeneic chondrocytes; 2) multi-layered scaffold with and without cells; and 3) randomly aligned scaffold with and without cells. After four months of follow-up, the outcome was assessed using histology and immunostaining. Results In vitro testing showed that the secreted ECM oriented itself along the fibre in multi-layered scaffolds. Both types of CG scaffolds supported cell viability, growth, and matrix synthesis. In vitro chondrogenesis on scaffold showed an around 400-fold increase in collagen type 2 (COL2A1) expression in both CG scaffolds, but the total glycosaminoglycan (GAG)/DNA deposition was 1.39-fold higher in the multi-layered scaffold than the randomly aligned scaffold. In vivo cartilage formation occurred in both multi-layered and randomly aligned scaffolds treated with and without cells, and was shown to be of hyaline phenotype on immunostaining. The defects treated with multi-layered + cells, however, showed significantly thicker cartilage formation than the randomly aligned scaffold. Conclusion We demonstrated that MSCs loaded CG scaffold with multi-layered zonal architecture promoted superior hyaline AC regeneration. Cite this article: Bone Joint Res 2020;9(9):601–612.

Electromagnetic fields enhance chondrogenesis of human adipose-derived stem cells in a chondrogenic microenvironment in vitro

2013 ◽  
Vol 114 (5) ◽  
pp. 647-655 ◽  
Author(s):  
Chung-Hwan Chen ◽  
Yi-Shan Lin ◽  
Yin-Chih Fu ◽  
Chih-Kuang Wang ◽  
Shun-Cheng Wu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Electromagnetic Field ◽  
Cell Viability ◽  
Chondrogenic Differentiation ◽  
Collagen Type ◽  
Cartilage Tissue ◽  
Pulse Electromagnetic Field ◽  
Type I ◽  
Adipose Derived Stem Cells ◽  
Pemf Treatment

We tested the hypothesis that electromagnetic field (EMF) stimulation enhances chondrogenesis in human adipose-derived stem cells (ADSCs) in a chondrogenic microenvironment. A two-dimensional hyaluronan (HA)-coated well (2D-HA) and a three-dimensional pellet culture system (3D-pellet) were used as chondrogenic microenvironments. The ADSCs were cultured in 2D-HA or 3D-pellet, and then treated with clinical-use pulse electromagnetic field (PEMF) or the innovative single-pulse electromagnetic field (SPEMF) stimulation. The cytotoxicity, cell viability, and chondrogenic and osteogenic differentiations were analyzed after PEMF or SPEMF treatment. The modules of PEMF and SPEMF stimulations used in this study did not cause cytotoxicity or alter cell viability in ADSCs. Both PEMF and SPEMF enhanced the chondrogenic gene expression (SOX-9, collagen type II, and aggrecan) of ADSCs cultured in 2D-HA and 3D-pellet. The expressions of bone matrix genes (osteocalcin and collagen type I) of ADSCs were not changed after SPEMF treatment in 2D-HA and 3D-pellet; however, they were enhanced by PEMF treatment. Both PEMF and SPEMF increased the cartilaginous matrix (sulfated glycosaminoglycan) deposition of ADSCs. However, PEMF treatment also increased mineralization of ADSCs, but SPEMF treatment did not. Both PEMF and SPEMF enhanced chondrogenic differentiation of ADSCs cultured in a chondrogenic microenvironment. SPEMF treatment enhanced ADSC chondrogenesis, but not osteogenesis, when the cells were cultured in a chondrogenic microenvironment. However, PEMF enhanced both osteogenesis and chondrogenesis under the same conditions. Thus the combination of a chondrogenic microenvironment with SPEMF stimulation can promote chondrogenic differentiation of ADSCs and may be applicable to articular cartilage tissue engineering.

scholarly journals Immunochemical and Mechanical Characterization of Cartilage Subtypes in Rabbit

2002 ◽  
Vol 50 (8) ◽  
pp. 1049-1058 ◽  
Author(s):  
Andreas Naumann ◽  
James E. Dennis ◽  
Amad Awadallah ◽  
David A. Carrino ◽  
Joseph M. Mansour ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Joint Analysis ◽  
Type I ◽  
Auricular Cartilage ◽  
Collagen Types ◽  
Type Vi Collagen ◽  
Intense Staining ◽  
Nasal Cartilage

Cartilage is categorized into three general subgroups, hyaline, elastic, and fibrocartilage, based primarily on morphologic criteria and secondarily on collagen (Types I and II) and elastin content. To more precisely define the different cartilage subtypes, rabbit cartilage isolated from joint, nose, auricle, epiglottis, and meniscus was characterized by immunohistochemical (IHC) localization of elastin and of collagen Types I, II, V, VI, and X, by biochemical analysis of total glycosaminoglycan (GAG) content, and by biomechanical indentation assay. Toluidine blue staining and safranin-O staining were used for morphological assessment of the cartilage subtypes. IHC staining of the cartilage samples showed a characteristic pattern of staining for the collagen antibodies that varied in both location and intensity. Auricular cartilage is discriminated from other subtypes by interterritorial elastin staining and no staining for Type VI collagen. Epiglottal cartilage is characterized by positive elastin staining and intense staining for Type VI collagen. The unique pattern for nasal cartilage is intense staining for Type V collagen and collagen X, whereas articular cartilage is negative for elastin (interterritorially) and only weakly positive for collagen Types V and VI. Meniscal cartilage shows the greatest intensity of staining for Type I collagen, weak staining for collagens V and VI, and no staining with antibody to collagen Type X. Matching cartilage samples were categorized by total GAG content, which showed increasing total GAG content from elastic cartilage (auricle, epiglottis) to fibrocartilage (meniscus) to hyaline cartilage (nose, knee joint). Analysis of aggregate modulus showed nasal and auricular cartilage to have the greatest stiffness, epiglottal and meniscal tissue the lowest, and articular cartilage intermediate. This study illustrates the differences and identifies unique characteristics of the different cartilage subtypes in rabbits. The results provide a baseline of data for generating and evaluating engineered repair cartilage tissue synthesized in vitro or for post-implantation analysis.

scholarly journals Hydrogels for the Application of Articular Cartilage Tissue Engineering: A Review of Hydrogels

2018 ◽  
Vol 2018 ◽  
pp. 1-13 ◽  
Author(s):  
Er-Yuan Chuang ◽  
Chih-Wei Chiang ◽  
Pei-Chun Wong ◽  
Chih-Hwa Chen
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Cartilage Damage ◽  
Medical Science ◽  
Cartilage Tissue Engineering ◽  
Polymeric Materials ◽  
Tissue Growth ◽  
Cartilage Tissue ◽  
Cellular Interaction ◽  
Polymeric Hydrogels

The treatment of articular cartilage damage is a major task in the medical science of orthopedics. Hydrogels possess the ability to form multifunctional cartilage grafts since they possess polymeric swellability upon immersion in an aqueous phase. Polymeric hydrogels are capable of physiological swelling and greasing, and they possess the mechanical behavior required for use as articular cartilage substitutes. The chondrogenic phenotype of these materials may be enhanced by embedding living cells. Artificial hydrogels fabricated from biologically derived and synthesized polymeric materials are also used as tissue-engineering scaffolds; with their controlled degradation profiles, the release of stimulatory growth factors can be achieved. In order to make use of these hydrogels, cartilage implants were formulated in the laboratory to demonstrate the bionic mechanical behaviors of physiological cartilage. This paper discusses developments concerning the use of polymeric hydrogels for substituting injured cartilage tissue and assisting tissue growth. These gels are designed with consideration of their polymeric classification, mechanical strength, manner of biodegradation, limitations of the payload, cellular interaction, amount of cells in the 3D hydrogel, sustained release for the model drug, and the different approaches for incorporation into adjacent organs. This article also summarizes the different advantages, disadvantages, and the future prospects of hydrogels.

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