scholarly journals An Instrumented Bioreactor for Mechanical Stimulation and Real-Time, Nondestructive Evaluation of Engineered Cartilage Tissue

2012 ◽  
Vol 6 (2) ◽  
Author(s):  
Jenni R. Popp ◽  
Justine J. Roberts ◽  
Doug V. Gallagher ◽  
Kristi S. Anseth ◽  
Stephanie J. Bryant ◽  
...  
Keyword(s):  
Nondestructive Evaluation ◽  
Mechanical Stimulation ◽  
Ultrasonic Transducer ◽  
Testing Machine ◽  
Cartilage Tissue ◽  
Matrix Deposition ◽  
Intermittent Compression ◽  
Engineered Tissue ◽  
Engineered Cartilage

Mechanical stimulation is essential for chondrocyte metabolism and cartilage matrix deposition. Traditional methods for evaluating developing tissue in vitro are destructive, time consuming, and expensive. Nondestructive evaluation of engineered tissue is promising for the development of replacement tissues. Here we present a novel instrumented bioreactor for dynamic mechanical stimulation and nondestructive evaluation of tissue mechanical properties and extracellular matrix (ECM) content. The bioreactor is instrumented with a video microscope and load cells in each well to measure tissue stiffness and an ultrasonic transducer for evaluating ECM content. Chondrocyte-laden hydrogel constructs were placed in the bioreactor and subjected to dynamic intermittent compression at 1 Hz and 10% strain for 1 h, twice per day for 7 days. Compressive modulus of the constructs, measured online in the bioreactor and offline on a mechanical testing machine, did not significantly change over time. Deposition of sulfated glycosaminoglycan (sGAG) increased significantly after 7 days, independent of loading. Furthermore, the relative reflection amplitude of the loaded constructs decreased significantly after 7 days, consistent with an increase in sGAG content. This preliminary work with our novel bioreactor demonstrates its capabilities for dynamic culture and nondestructive evaluation.

scholarly journals DMOG Negatively Impacts Tissue Engineered Cartilage Development

Cartilage ◽  
2020 ◽  
pp. 194760352096706
Author(s):  
Jessica M. Falcon ◽  
Dylan Chirman ◽  
Alyssa Veneziale ◽  
Justin Morman ◽  
Katherine Bolten ◽  
...  
Keyword(s):  
Positive Impact ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Short Term ◽  
Short Term Treatment ◽  
Matrix Deposition ◽  
Matrix Production ◽  
Engineered Cartilage ◽  
The Impact

Objective Articular cartilage exists in a hypoxic environment, which motivates the use of hypoxia-simulating chemical agents to improve matrix production in cartilage tissue engineering. The aim of this study was to investigate whether dimethyloxalylglycine (DMOG), a HIF-1α stabilizer, would improve matrix production in 3-dimensional (3D) porcine synovial-derived mesenchymal stem cell (SYN-MSC) co-culture with chondrocytes. Design Pellet cultures and scaffold-based engineered cartilage were grown in vitro to determine the impact of chemically simulated hypoxia on 2 types of 3D cell culture. DMOG-treated groups were exposed to DMOG from day 14 to day 21 and grown up to 6 weeks with n = 3 per condition and time point. Results The addition of DMOG resulted in HIF-1α stabilization in the exterior of the engineered constructs, which resulted in increased regional type II collagen deposition, but the stabilization did not translate to overall increased extracellular matrix deposition. There was no increase in HIF-1α stabilization in the pellet cultures. DMOG treatment also negatively affected the mechanical competency of the engineered cartilage. Conclusions Despite previous studies that demonstrated the efficacy of DMOG, here, short-term treatment with DMOG did not have a uniformly positive impact on the chondrogenic capacity of SYN-MSCs in either pellet culture or in scaffold-based engineered cartilage, as evidenced by reduced matrix production. Such 3D constructs generally have a naturally occurring hypoxic center, which allows for the stabilization of HIF-1α in the interior tissue. Thus, short-term addition of DMOG may not further improve this in cartilage tissue engineered constructs.

Fabrication of Scaffold-Free Engineered Cartilage Using Passaged Chondrocytes in the Presence of Insuline Like Growth Factor 1 (IGF-1)

2007 ◽  
Vol 342-343 ◽  
pp. 149-152 ◽  
Author(s):  
Ri Long Jin ◽  
So Ra Park ◽  
Jeong Hwa Son ◽  
Byoung Hyun Min
Keyword(s):  
Growth Factor ◽  
Articular Chondrocytes ◽  
Cartilage Tissue ◽  
Insulin Like Growth Factor ◽  
Free System ◽  
Engineered Cartilage

Two passaged (P2) immature porcine articular chondrocytes were used to fabricate an engineered cartilage tissue in an in vitro scaffold-free system with or without insulin like growth factor 1 (IGF-1). This study shows the possibility of the fabrication of structurally regular neocartilage tissue using passaged chondrocytes in the scaffold-free system with insulin like growth factor-1(IGF-1).

Directing Chondrogenesis of Primary Chondrocytes by Exposure to Glucose Concentrations

2015 ◽  
Vol 24 ◽  
pp. 30-42
Author(s):  
Samuel C. Uzoechi ◽  
Kennedy O. Ejeta ◽  
Goddy C. Okoye ◽  
Gideon I. Ndubuka ◽  
Patrick Ugochukwu Agbasi ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Glucose Concentration ◽  
Cartilage Regeneration ◽  
Cartilage Tissue ◽  
Glycosaminoglycan Synthesis ◽  
Chondrogenic Medium ◽  
Hypoxic Conditions ◽  
Matrix Production ◽  
Engineered Cartilage

Since articular cartilage is avascular, both nutrient supply and metabolic waste excretion depend on diffusion. However, the major cause of the progression of articular cartilage defect is the poor inherent regenerative capacity of chondrocytes which limits the process of cartilage tissue repair. Creation of nutrient gradients in in vitro cell culture, however, can provide a clue on zonal distributions of cells and glycosaminoglycan synthesis throughout the tissue engineered cartilage. We hypothesized that glucose gradient, in combination with growth factors, could induce differences in matrix distributions for articular cartilage regeneration. Chondrocytes were harvested from bovine cartilage and expanded in monolayers. First, either p0 or p2 chondrocytes were differentiated in serum-free chondrogenic medium containing different glucose concentrations supplemented with TGFβ3/dex or IGF-1under hypoxic or normoxic conditions for 7 days in monolayer. The results indicate that cellular metabolism, cell numbers and glycosaminoglycan (GAG) content increased with increase in glucose concentration in all conditions. Aggrecan (AGC) expression consistently increased with decreasing glucose concentration in both normoxic and hypoxic conditions. COL II and COL I expressions increased with increasing glucose concentration up to 5mmol/L. The expression of COMP increased with increasing glucose concentration under hypoxic conditions and interestingly showed an opposite trend under normoxic conditions. However, comparing the chondrogenic capacity of p0 and p2 cells in the different glucose concentrations did not show differences, but the potential of p2 cells was in general lower compared to p0. Hypoxia had stimulatory effects on matrix production compared to normoxia in both passages. Therefore, supplemented glucose concentration in monolayer could induce differences in matrix production, but the chondrogenic potential remained equal. Therefore, this information could be use to a create gradients through a tissue-engineered cartilage.

scholarly journals Effects of bionic mechanical stimulation on the properties of engineered cartilage tissue

2020 ◽  
Author(s):  
Zhiyan Hao ◽  
Sen Wang ◽  
Jichang Nie ◽  
Dichen Li ◽  
Ao Fang ◽  
...  
Keyword(s):  
Mechanical Stimulation ◽  
Cartilage Tissue ◽  
Engineered Cartilage

scholarly journals Physioxia stimulates extra-cellular matrix deposition and increases mechanical properties of human chondrocyte-derived tissue-engineered cartilage

2020 ◽  
Author(s):  
J E Dennis ◽  
G A Whitney ◽  
J Rai ◽  
R J Fernandes ◽  
T J Kean
Keyword(s):  
Extracellular Matrix ◽  
Oxygen Tension ◽  
Atmospheric Oxygen ◽  
Cartilage Tissue ◽  
Type Ii ◽  
Matrix Deposition ◽  
Extracellular Matrix Components ◽  
Cross Links ◽  
Matrix Components ◽  
Engineered Cartilage

AbstractCartilage tissue has been recalcitrant to tissue engineering approaches. In this study, human chondrocytes were formed into self-assembled cartilage sheets, cultured in physiologic (5%) and atmospheric (20%) oxygen conditions and underwent biochemical, histological and biomechanical analysis at one- and two-months. The results indicated that sheets formed at physiological oxygen tension were thicker, contained greater amounts of glycosaminoglycans (GAGs) and type II collagen, and had greater compressive and tensile properties than those cultured in atmospheric oxygen. In all cases, cartilage sheets stained throughout for extracellular matrix components. Type II-IX-XI collagen heteropolymer formed in the neo-cartilage and fibrils were stabilized by trivalent pyridinoline cross-links. Collagen cross-links were not significantly affected by oxygen tension but increased with time in culture. Physiological oxygen tension and longer culture periods both served to increase extracellular matrix components. The foremost correlation was found between compressive stiffness and the GAG to collagen ratio.SummaryTissue-engineered cartilage formed from human articular chondrocytes produces thicker, stiffer, more extracellular-matrix rich cartilage tissue when grown under physiological (5%) vs. atmospheric oxygen (20%) tension.

scholarly journals The Effect of Varying Concentrations and Application Periods of Chondroitinase ABC on Tissue-Engineered Cartilage

2014 ◽  
Vol 1 (1) ◽  
Author(s):  
Eric Tong ◽  
Grace D. O'Connell ◽  
Terri-Ann N. Kelly ◽  
Clark T. Hung
Keyword(s):  
Mechanical Properties ◽  
Articular Cartilage ◽  
Treatment Option ◽  
Type Ii Collagen ◽  
Treated Group ◽  
Cartilage Tissue ◽  
Type Ii ◽  
Chondroitinase Abc ◽  
Engineered Cartilage

Osteoarthritis, a chronic malady characterized by joint pain and swelling, is caused by damage to articular cartilage and is perpetuated by low-grade inflammation.  Treatments for osteoarthritis do exist, but many treatments focus on coping with the disease rather than curing it.  Surgical options that replace damaged cartilage tissue with that of donor cartilage tissue or cartilage tissue from other parts of articular joints face complications especially when the tissue is not of the correct size or does not have native-like properties. A more suitable treatment option for osteoarthritis is to develop an in vitro tissue-engineered cartilage construct that can be grown using the patient’s own cells and to surgically remove the patient’s damaged cartilage and replace it with the tissue-engineered cartilage. A challenge in developing such a treatment option is producing tissue-engineered cartilage with mechanical properties akin to those of native human articular cartilage. This challenge may be overcome by maximizing the production of type II collagen by the chondrocytes in vitro. One way to maximize collagen production is through the application of chondroitinase ABC, an enzyme which temporarily suppresses proteoglycans in the cartilage matrix to create more space for type II collagen to develop. In this study, two two levels of cABC treatment were applied (“high” and “low”) to cartilage tissue constructs. The “low” cABC treated group received daily feeding of 0.075 U/mL from day 14 to 21 followed by a replacement of chondrogenic media without cABC.  The “high” cABC treated group received a single addition of 0.15 U/mL from day 14 to 16 followed by a replacement of chondrogenic media without cABC.  At the end of 42 days, the constructs were subjected to mechanical testing and biochemical analyses. These analyses showed that the high cABC treatment yielded more native-like mechanical properties when compared to the low cABC treatment and the control results.  Biochemical and histological analyses confirmed that the proteoglycan and collagen II content were higher in the low and high cABC treated groups when compared to the control. All analyses show that the most efficient application of chondroitinase ABC is through a two day duration treatment of a higher concentration (0.15 U/mL).

Bioprinting of biomimetic self-organised cartilage with a supporting joint fixation device

2021 ◽  
Vol 14 (1) ◽  
pp. 015008
Author(s):  
Ross Burdis ◽  
Farhad Chariyev-Prinz ◽  
Daniel J Kelly
Keyword(s):  
Complete Solution ◽  
Cartilage Tissue ◽  
Synovial Joint ◽  
Fixation Device ◽  
Collagen Network ◽  
Parallel Fibre ◽  
Engineered Tissue ◽  
Self Organisation ◽  
Engineered Cartilage

Abstract Despite sustained efforts, engineering truly biomimetic articular cartilage (AC) via traditional top-down approaches remains challenging. Emerging biofabrication strategies, from 3D bioprinting to scaffold-free approaches that leverage principles of cellular self-organisation, are generating significant interest in the field of cartilage tissue engineering as a means of developing biomimetic tissue analogues in vitro. Although such strategies have advanced the quality of engineered cartilage, recapitulation of many key structural features of native AC, in particular a collagen network mimicking the tissue’s ‘Benninghoff arcade’, remains elusive. Additionally, a complete solution to fixating engineered cartilages in situ within damaged synovial joints has yet to be identified. This study sought to address both of these key challenges by engineering biomimetic AC within a device designed to anchor the tissue within a synovial joint defect. We first designed and fabricated a fixation device capable of anchoring engineered cartilage into the subchondral bone. Next, we developed a strategy for inkjet printing porcine mesenchymal stem/stromal cells (MSCs) into this supporting fixation device, which was also designed to provide instructive cues to direct the self-organisation of MSC condensations towards a stratified engineered AC. We found that a higher starting cell-density supported the development of a more zonally defined collagen network within the engineered tissue. Dynamic culture was implemented to further enhance the quality of this engineered tissue, resulting in an approximate 3 fold increase in glycosaminoglycan and collagen accumulation. Ultimately this strategy supported the development of AC that exhibited near-native levels of glycosaminoglycan accumulation (>5% WW), as well as a biomimetic collagen network organisation with a perpendicular to a parallel fibre arrangement (relative to the tissue surface) from the deep to superficial zones via arcading fibres within the middle zone of the engineered tissue. Collectively, this work demonstrates the successful convergence of novel biofabrication methods, bioprinting strategies and culture regimes to engineer a hybrid implant suited to resurfacing AC defects.

scholarly journals Human adipose-derived stromal/stem cells expressing doublecortin improve cartilage repair in rabbits and monkeys

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Dongxia Ge ◽  
Michael J. O’Brien ◽  
Felix H. Savoie ◽  
Jeffrey M. Gimble ◽  
Xiying Wu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cartilage Repair ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Cartilage Lesions ◽  
Tissue Constructs ◽  
Relieve Pain ◽  
Engineered Cartilage ◽  
Stromal Stem Cells

AbstractLocalized cartilage lesions in early osteoarthritis and acute joint injuries are usually treated surgically to restore function and relieve pain. However, a persistent clinical challenge remains in how to repair the cartilage lesions. We expressed doublecortin (DCX) in human adipose-derived stromal/stem cells (hASCs) and engineered hASCs into cartilage tissues using an in vitro 96-well pellet culture system. The cartilage tissue constructs with and without DCX expression were implanted in the knee cartilage defects of rabbits (n = 42) and monkeys (n = 12). Cohorts of animals were euthanized at 6, 12, and 24 months after surgery to evaluate the cartilage repair outcomes. We found that DCX expression in hASCs increased expression of growth differentiation factor 5 (GDF5) and matrilin 2 in the engineered cartilage tissues. The cartilage tissues with DCX expression significantly enhanced cartilage repair as assessed macroscopically and histologically at 6, 12, and 24 months after implantation in the rabbits and 24 months after implantation in the monkeys, compared to the cartilage tissues without DCX expression. These findings suggest that hASCs expressing DCX may be engineered into cartilage tissues that can be used to treat localized cartilage lesions.

scholarly journals CARTILAGE TISSUE ENGINEERING

2005 ◽  
Vol 17 (02) ◽  
pp. 61-71 ◽  
Author(s):  
CHIH-HUNG CHANG ◽  
FENG-HUEI LIN ◽  
TZONG-FU KUO ◽  
HWA-CHANG LIU
Keyword(s):  
Tissue Engineering ◽  
Animal Studies ◽  
Cartilage Tissue Engineering ◽  
In Vivo Studies ◽  
Cartilage Tissue ◽  
Current Status ◽  
Materials Used ◽  
Engineered Cartilage

Tissue engineering is a new approach for articular cartilage repair. The aim of the present article was to review the current status of cartilage tissue engineering researches. The scaffold materials used for cartilage tissue engineering, the in vitro, in vivo studies and the clinical trials were all reviewed. Our researches about in vitro cartilage tissue engineering with new type bioactive scaffold and preliminary animal studies results will also be described. The scaffold was tricopolymer made from gelatin, hyaluronan and chondroitin. Chondrocytes seeded in tricopolymer showed in vitro engineered cartilage formation. The engineered cartilage constructs were implanted into knee joints of miniature pigs for animal study.

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