scholarly journals Effects of Serum and Compressive Loading on the Cartilage Matrix Synthesis and Spatiotemporal Deposition Around Chondrocytes in 3D Culture

2013 ◽  
Vol 19 (9-10) ◽  
pp. 1199-1208 ◽  
Author(s):  
Peihui Wu ◽  
Elizabeth DeLassus ◽  
Debabrata Patra ◽  
Weiming Liao ◽  
Linda J. Sandell
Keyword(s):  
Compressive Loading ◽  
3D Culture ◽  
Cartilage Matrix ◽  
Matrix Synthesis

scholarly journals Low Magnitude of Compression Enhances Biosynthesis of Mesenchymal Stem Cells towards Nucleus Pulposus Cells via the TRPV4-Dependent Pathway

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Yibo Gan ◽  
Bing Tu ◽  
Pei Li ◽  
Jixing Ye ◽  
Chen Zhao ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Transient Receptor Potential ◽  
Compressive Loading ◽  
3D Culture ◽  
Receptor Potential ◽  
Immunohistochemical Analysis ◽  
Transient Receptor Potential Vanilloid ◽  
Nucleus Pulposus Cells ◽  
Anabolic Response ◽  
Low Magnitude

Mesenchymal stem cell- (MSC-) based therapy is regarded as a promising tissue engineering strategy to achieve nucleus pulposus (NP) regeneration for the treatment of intervertebral disc degeneration (IDD). However, it is still a challenge to promote the biosynthesis of MSC to meet the requirement of NP regeneration. The purpose of this study was to optimize the compressive magnitude to enhance the extracellular matrix (ECM) deposition towards discogenesis of MSCs. Thus, we constructed a 3D culture model for MSCs to bear different magnitudes of compression for 7 days (5%, 10%, and 20% at the frequency of 1.0 Hz for 8 hours/day) using an intelligent and mechanically active bioreactor. Then, the underlying mechanotransduction mechanism of transient receptor potential vanilloid 4 (TRPV4) was further explored. The MSC-encapsulated hybrids were evaluated by Live/Dead staining, biochemical content assay, real-time PCR, Western blot, histological, and immunohistochemical analysis. The results showed that low-magnitude compression promoted anabolic response where high-magnitude compression induced the catabolic response for the 3D-cultured MSCs. The anabolic effect of low-magnitude compression could be inhibited by inhibiting TRPV4. Meanwhile, the activation of TRPV4 enhanced the biosynthesis analogous to low-magnitude compression. These findings demonstrate that low-magnitude compression promoted the anabolic response of ECM deposition towards discogenesis for the 3D-cultured MSCs and the TRPV4 channel plays a key role on mechanical signal transduction for low-magnitude compressive loading. Further understanding of this mechanism may provide insights into the development of new therapies for MSC-based NP regeneration.

Repeated Dynamic Loading Modulates Cartilage Gene Expression but Does Not Improve Mechanical Properties of MSC-Laden Hydrogels

2009 ◽  
Author(s):  
Alice H. Huang ◽  
Robert L. Mauck
Keyword(s):  
Gene Expression ◽  
Mechanical Properties ◽  
Dynamic Compression ◽  
Compressive Loading ◽  
3D Culture ◽  
Tissue Growth ◽  
Cartilage Tissue ◽  
Seeding Density ◽  
Porous Scaffolds ◽  
Positive Effects

Mesenchymal stem cells (MSCs) are a multi-potential cell type that can differentiate toward a variety of tissue-specific phenotypes, including cartilage. Given their chondrogenic potential, MSCs are a promising cell source for cartilage tissue engineering (TE). However, while MSCs readily undergo chondrogenesis in 3D culture and deposit a cartilage-like matrix, the mechanical properties of MSC-seeded constructs are greatly inferior to chondrocyte-seeded constructs similarly maintained [1]. To date, optimization strategies for enhancing functional MSC chondrogenesis, including increasing seeding density and transient application of growth factor, have shown limited success [3]. Using microarray analysis, we have recently demonstrated that mis-expression of certain genes, including lubricin, chondromodulin and RGD-CAP, a collagen associated protein, may underlie this disparity in mechanical function [2]. In this study, we examined dynamic compression as an alternative method to enhance MSC differentiation. Previous work using chondrocyte-based constructs have demonstrated that matrix biosynthesis and mechanical properties were improved with the application of cyclic compression [4]. Furthermore, upregulation of lubricin was observed when surface motion was applied to chondrocyte-seeded porous scaffolds [5]. While significant effort has gone toward optimizing loading parameters to direct tissue growth of chondrocyte-based constructs, few studies have examined the effects of mechanical stimulation on MSC-based constructs. Some have demonstrated positive effects on MSC chondrogenesis with application of compressive loading [6, 7], while others have shown that long-term loading may adversely affect the developing mechanical properties of MSC-seeded constructs [8]. In this study, we examined the effects of repeated dynamic compressive loading on MSC chondrogenesis and showed that mechanical properties and gene expression were modulated by this loading modality.

Eel insulin: isolation, characterization and stimulatory actions on [35S]sulphate and [3H]thymidine uptake in the branchial cartilage of the eel in vitro

1992 ◽  
Vol 133 (2) ◽  
pp. 221-230 ◽  
Author(s):  
C. Duan ◽  
T. Noso ◽  
S. Moriyama ◽  
H. Kawauchi ◽  
T. Hirano
Keyword(s):  
Bovine Insulin ◽  
Cartilage Matrix ◽  
Common Mechanism ◽  
Thymidine Uptake ◽  
Dependent Manner ◽  
Matrix Synthesis ◽  
Sulphate Uptake ◽  
Igf I ◽  
Dose Dependent

ABSTRACT Our previous studies have shown that mammalian and salmon insulins stimulate sulphate uptake by cultured eel cartilage, suggesting the possible involvement of insulin in the regulation of cartilage matrix synthesis. In the present study, homologous eel insulin was isolated and characterized, and its effects on cartilage matrix synthesis and DNA synthesis were examined in vitro. Insulin was extracted from eel pancreas with acid–ethanol, and subsequently purified by isoelectric precipitation at pH 5·3, gel filtration on Sephadex G-50, and reversed-phase high-performance liquid chromatography. The amino acid composition and complete sequence (50 residues) of eel insulin revealed high homology to teleostean and mammalian insulins. The isolated eel insulin produced a more pronounced and longer lasting hypoglycaemic effect than bovine insulin in the eel. Homologous eel insulin, like bovine insulin-like growth factor (IGF-I) and insulin, stimulated sulphate uptake by cultured eel cartilage in a dose-dependent manner (16–1000 ng/ml). Combination experiments using maximal concentrations of bovine IGF-I (250 ng/ml) and increasing amounts of eel insulin (10–250 ng/ml) showed no additive effects of insulin on sulphate uptake, suggesting that insulin and IGF-I may share a common mechanism(s) of action. Eel insulin and bovine IGF-I also enhanced thymidine incorporation by eel cartilage in a dose-dependent manner (4–1000 ng/ml); eel insulin was equipotent with bovine IGF-I. These results suggest that insulin, like IGF-I, may exert direct growth-promoting actions in branchial cartilage of the eel. Journal of Endocrinology (1992) 133, 221–230

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