Effects of Mechanical Vibration on Matrix Production and Proliferation of Three-Dimensional Cultured Chondrocytes

2008 ◽  
Author(s):  
Yuta Takagi ◽  
Toshihiko Shiraishi ◽  
Shin Morishita ◽  
Ryohei Takeuchi ◽  
Tomoyuki Saito ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Signaling Pathways ◽  
Weight Bearing ◽  
Mechanical Vibration ◽  
Three Dimensional ◽  
Cartilage Tissue ◽  
Vibration Treatment ◽  
Matrix Production

This paper describes the effects of vibration stimulation on chondrocytes in three-dimensional culture in relation to the production of regenerative cartilage tissue, using collagen artificial skin as a carrier and supplementation with hyaluronic acid (used in the conservative treatment of osteoarthritis), and the mechanism of the adaptive response of chondrocytes to mechanical loading. The experimental condition imitates an environment of articular cartilage in vivo that chondrocytes are completely surrounded by the extracellular matrix and receives mechanical stimulation for the weight-bearing mechanics. Chondrocytes were isolated from articular cartilage of porcine metatarsophalangeal joints. Experiments were performed under four different culture conditions: control condition, in which chondrocytes were cultured with atelocollagen gel and collagen artificial skins, and no vibration (HA−Vib−); HA−Vib+, in which chondrocytes were cultured in atelocollagen gel and collagen artificial skins with vibration treatment for 2 weeks; HA+Vib−, in which chondrocytes were cultured in medium containing 0.1% hyaluronic acid; and HA+Vib+, in which chondrocytes were cultured in medium containing 0.1% hyaluronic acid with vibration treatment for 2 weeks. Histologic analysis was conducted at 14 days of culture. The proliferation of chondrocytes was obtained by counting the number of cells with a hemocytometer after 3, 7, 10, and 14 days of culture. The expression of Sox 9 and β-catenin was detected by western blotting analysis. Sox 9 has been reported of involvement in transcription of type IX collagen that binds cartilage-specific type II collagen fibrils. β-catenin plays an important role of signaling pathways of cell proliferation although the relationship between β-catenin and mechanical vibration stimulation has not been clarified yet. The obtained results are as follows. The mechanical vibration enhanced the thickness of extracellular matrix of chondrocytes in histologic section at 14 days of culture and increased the expression of Sox 9. In addition, the mechanical vibration significantly increased the number of chondrocytes after 10 days of culture and promoted the expression of β-catenin. These results show that mechanical vibration promotes the matrix production and proliferation of chondrocytes and that a part of important signaling pathways in relation to mechanical vibration stimulation and proliferation of chondrocytes has been revealed.

Tissue-Engineered Cartilage in Three-Dimensional Cultured Chondrocytes by Ultrasound Stimulation and Collagen Sponge as Scaffold

2009 ◽  
Author(s):  
Toshihiko Shiraishi ◽  
Ietomo Matsunaga ◽  
Shin Morishita ◽  
Ryohei Takeuchi ◽  
Tomoyuki Saito ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Three Dimensional ◽  
Collagen Sponge ◽  
Cartilage Tissue ◽  
Mechanical Environment ◽  
Ultrasound Stimulation ◽  
Scaffold Structure ◽  
Matrix Production ◽  
Engineered Cartilage ◽  
Cultured Chondrocytes

This paper describes the effects of ultrasound stimulation on chondrocytes in three-dimensional culture in relation to the production of regenerative cartilage tissue, using collagen sponges as a carrier and supplementation with hyaluronic acid (used in the conservative treatment of osteoarthritis). It has been shown that cell proliferation and matrix production can be facilitated by considering the mechanical environment of the cultured chondrocytes and the mechanical properties of the scaffold structure used in the culture and of the stimulation used.

scholarly journals Challenges in Fabrication of Tissue-Engineered Cartilage with Correct Cellular Colonization and Extracellular Matrix Assembly

2018 ◽  
Vol 19 (9) ◽  
pp. 2700 ◽  
Author(s):  
Mikko Lammi ◽  
Juha Piltti ◽  
Juha Prittinen ◽  
Chengjuan Qu
Keyword(s):  
Extracellular Matrix ◽  
Articular Cartilage ◽  
Pore Size ◽  
Cartilage Tissue ◽  
Cellular Interactions ◽  
Matrix Assembly ◽  
Small Pore ◽  
Culture Models ◽  
Open Question ◽  
Engineered Cartilage

A correct articular cartilage ultrastructure regarding its structural components and cellularity is important for appropriate performance of tissue-engineered articular cartilage. Various scaffold-based, as well as scaffold-free, culture models have been under development to manufacture functional cartilage tissue. Even decellularized tissues have been considered as a potential choice for cellular seeding and tissue fabrication. Pore size, interconnectivity, and functionalization of the scaffold architecture can be varied. Increased mechanical function requires a dense scaffold, which also easily restricts cellular access within the scaffold at seeding. High pore size enhances nutrient transport, while small pore size improves cellular interactions and scaffold resorption. In scaffold-free cultures, the cells assemble the tissue completely by themselves; in optimized cultures, they should be able to fabricate native-like tissue. Decellularized cartilage has a native ultrastructure, although it is a challenge to obtain proper cellular colonization during cell seeding. Bioprinting can, in principle, provide the tissue with correct cellularity and extracellular matrix content, although it is still an open question as to how the correct molecular interaction and structure of extracellular matrix could be achieved. These are challenges facing the ongoing efforts to manufacture optimal articular cartilage.

Zonal Poisson Effect on the Chondron Deformation in Articular Cartilage under Compression

2007 ◽  
Vol 342-343 ◽  
pp. 133-136
Author(s):  
Jae Bong Choi
Keyword(s):  
Extracellular Matrix ◽  
Articular Cartilage ◽  
Mechanical Response ◽  
Three Dimensional ◽  
Pericellular Matrix ◽  
Poisson Effect ◽  
Type Vi Collagen ◽  
Confocal Images ◽  
Three Dimensional Models

The objective of this study was to quantify the zonal difference of the in situ chondron’s Poisson effect under different magnitudes of compression. Fluorescence immunolabeling for type VI collagen was used to identify the pericellular matrix (PCM) and chondron, and a series of fluorescent confocal images were recorded and reconstructed to form quantitative three-dimensional models. The zonal variations in the mechanical response of the chondron do not appear to be due to zonal differences in PCM properties, but rather seem to result from significant inhomogeneities in relative stiffnesses of the extracellular matrix (ECM) and PCM with depth.

scholarly journals A biomimetic extracellular matrix for cartilage tissue engineering centered on photocurable gelatin, hyaluronic acid and chondroitin sulfate

2014 ◽  
Vol 10 (1) ◽  
pp. 214-223 ◽  
Author(s):  
Peter A. Levett ◽  
Ferry P.W. Melchels ◽  
Karsten Schrobback ◽  
Dietmar W. Hutmacher ◽  
Jos Malda ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Chondroitin Sulfate ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue

scholarly journals Localization of hyaluronic acid in human articular cartilage.

1994 ◽  
Vol 42 (4) ◽  
pp. 513-522 ◽  
Author(s):  
A Asari ◽  
S Miyauchi ◽  
S Kuriyama ◽  
A Machida ◽  
K Kohno ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Dermatan Sulfate ◽  
Keratan Sulfate ◽  
Human Articular Cartilage ◽  
Middle Zone ◽  
Deep Zone ◽  
Weak Staining ◽  
Pre Treatment

To demonstrate localization of hyaluronic acid (HA) in articular cartilage of the human femur, biotinylated HA-binding region, which specifically binds HA molecules, was applied to the tissue. In sections fixed by 2% paraformaldehyde-2% glutaraldehyde, HA staining was detected in lamina splendens and chondrocytes in the middle zone. By pretreatment with trypsin, intense HA staining appeared in the extracellular matrix of the deep zone and weak staining in the superficial and middle zones. Moreover, pre-treatment with chondroitinase ABC (CHase ABC) intensely enhanced the stainability for HA in the superficial and middle zones and weakly in the deeper zone. Combined pre-treatment of trypsin with CHase ABC abolished intra- and extracellular staining for HA in all zones. By microbiochemical study, the concentrations of HA and dermatan sulfate were high in the middle zone, whereas those of chondroitin sulfate and keratan sulfate were high in the deep zone. These results suggest that HA is abundantly synthesized in and secreted from the chondrocytes, particularly in the middle zone, whereas it is largely masked by proteoglycan constituents in the extracellular matrix.

scholarly journals TISSUE ENGINEERING: FROM RESEARCH TO CLINICAL APPLICATION IN REGENERATIVE MEDICINE

1970 ◽  
pp. 159-192
Author(s):  
Enrico Tognana ◽  
Lanfranco Callegaro
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Regenerative Medicine ◽  
Therapeutic Option ◽  
Cartilage Tissue ◽  
Sequence Of Events ◽  
Benzyl Esters ◽  
Tissue Restoration

Tissue engineering strategies have recently emerged as the most advanced therapeutic option presently available in regenerative medicine. Tissue engineering encompasses the use of cells and their molecules in artificial constructs that compensate for lost or impaired body functions. It is based upon scaffoldguided tissue regeneration and involves the seeding of porous, biodegradable scaffolds with donor cells, which become differentiated and mimic naturally occurring tissues. These tissue-engineered constructs are then implanted into the patient to replace diseased or damaged tissues. Our approach to regenerative medicine is based on hyaluronan derivative polymers. HYAFF® is a class of hyaluronan derivative polymers obtained by coupling reaction. The strategy behind the creation of these polymers was to improve the stability of the polymer by esterifying the free carboxyl group of glucuronic acid, frequently repeated along the hyaluronic acid chain, with different types of alcohols. Once esterification of the polymer has been obtained, the material can easily be processed to produce membranes, fibres, sponges, microspheres and other devices, by extrusion, lyophilization or spray drying. A broad variety of polymers can be subsequently generated either by changing the type of ester group introduced or the extent of the esterification. The benzyl esters of hyaluronan, termed HYAFF®-11, are one of the most characterized HYAFF® polymers, from both the physicochemical and biological viewpoints, produced starting from hyaluronan of about 200 KDa. The ideal scaffold for tissue engineering should provide an immediate support to cells and have mechanical properties matching those of the tissue being repaired. Gradually then the material should be resorbed, as the cells begin secreting their own extracellular matrix, thus allowing for an optimal integration between newformed and existing tissue. Extensive biocompatibility studies have demonstrated the safety of HYAFF® scaffolds and their ability to be resorbed in the absence of an inflammatory response. Moreover, when implanted tend to promote the recapitulation of the events that facilitate tissue repair. HYAFF®-11 three-dimensional matrices support the in vitro growth of highly viable chondrocytes and fibroblasts. Similarly, micro-perforated membrane supports the growth and differentiation of keratinocytes. These cells, previously expanded on plastic and hence seeded into the HYAFF® scaffold, produce a characteristic extracellular matrix rich in proteoglycans expressing the typical markers of the tissues of their origin. Hyaluronan presents a variety of multi-functional activity being both a structural and informational molecule. Investigation of hyaluronan synthesis and degradation, the identification of new receptors and binding proteins and the elucidation of hyaluronan-dependent signaling pathways keep providing novel insights into the true biological functions of this intriguing polymer. The possibility to elaborate this natural polymer in different physical forms, as HYAFF® biopolymers family is allowing to do, has given the opportunity to translate tissue engineering strategies in clinical practice providing a biomaterial that induces and modulates the sequence of events that lead to damage tissue restoration. The following chapter will report how tissue engineering approach and hyaluronic acid technology could improve the biological function of cell transplantation in the treatment of tissue defects, in particular for skin and cartilage tissue restoration.

Hyaluronic acid-based hydrogels to study cancer cell behaviors

2021 ◽  
Author(s):  
Kasra Goodarzi ◽  
Shreyas Rao
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Cancer Cell ◽  
Three Dimensional ◽  
Cell Behaviors ◽  
Natural Polysaccharide

Hyaluronic acid (HA) is a natural polysaccharide and a key component of the extracellular matrix (ECM) in many tissues. Therefore, HA-based biomaterials are extensively utilized to create three dimensional ECM...

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.

Cell-derived decellularized extracellular matrix scaffolds for articular cartilage repair

2020 ◽  
pp. 039139882095386
Author(s):  
Wenrun Zhu ◽  
Lu Cao ◽  
Chunfeng Song ◽  
Zhiying Pang ◽  
Haochen Jiang ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Articular Cartilage ◽  
Cartilage Repair ◽  
Cell Attachment ◽  
Tissue Growth ◽  
Bioactive Molecules ◽  
Cartilage Tissue ◽  
Articular Cartilage Repair ◽  
Decellularized Extracellular Matrix

Articular cartilage repair remains a great clinical challenge. Tissue engineering approaches based on decellularized extracellular matrix (dECM) scaffolds show promise for facilitating articular cartilage repair. Traditional regenerative approaches currently used in clinical practice, such as microfracture, mosaicplasty, and autologous chondrocyte implantation, can improve cartilage repair and show therapeutic effect to some degree; however, the long-term curative effect is suboptimal. As dECM prepared by proper decellularization procedures is a biodegradable material, which provides space for regeneration tissue growth, possesses low immunogenicity, and retains most of its bioactive molecules that maintain tissue homeostasis and facilitate tissue repair, dECM scaffolds may provide a biomimetic microenvironment promoting cell attachment, proliferation, and chondrogenic differentiation. Currently, cell-derived dECM scaffolds have become a research hotspot in the field of cartilage tissue engineering, as ECM derived from cells cultured in vitro has many advantages compared with native cartilage ECM. This review describes cell types used to secrete ECM, methods of inducing cells to secrete cartilage-like ECM and decellularization methods to prepare cell-derived dECM. The potential mechanism of dECM scaffolds on cartilage repair, methods for improving the mechanical strength of cell-derived dECM scaffolds, and future perspectives on cell-derived dECM scaffolds are also discussed in this review.

scholarly journals Label-free Raman monitoring of extracellular matrix formation in three-dimensional polymeric scaffolds

2013 ◽  
Vol 10 (86) ◽  
pp. 20130464 ◽  
Author(s):  
Aliz Kunstar ◽  
Anne M. Leferink ◽  
Paul I. Okagbare ◽  
Michael D. Morris ◽  
Blake J. Roessler ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Raman Spectroscopy ◽  
Three Dimensional ◽  
Raman Microspectroscopy ◽  
Collagen Content ◽  
Cartilage Tissue ◽  
Fibre Optic ◽  
Label Free ◽  
Tissue Quality ◽  
Non Invasive

Monitoring extracellular matrix (ECM) components is one of the key methods used to determine tissue quality in three-dimensional scaffolds for regenerative medicine and clinical purposes. Raman spectroscopy can be used for non-invasive sensing of cellular and ECM biochemistry. We have investigated the use of conventional (confocal and semiconfocal) Raman microspectroscopy and fibre-optic Raman spectroscopy for in vitro monitoring of ECM formation in three-dimensional poly(ethylene oxide terephthalate)–poly(butylene terephthalate) (PEOT/PBT) scaffolds. Chondrocyte-seeded PEOT/PBT scaffolds were analysed for ECM formation by Raman microspectroscopy, biochemical analysis, histology and scanning electron microscopy. ECM deposition in these scaffolds was successfully detected by biochemical and histological analysis and by label-free non-destructive Raman microspectroscopy. In the spectra collected by the conventional Raman set-ups, the Raman bands at 937 and at 1062 cm −1 which, respectively, correspond to collagen and sulfated glycosaminoglycans could be used as Raman markers for ECM formation in scaffolds. Collagen synthesis was found to be different in single chondrocyte-seeded scaffolds when compared with microaggregate-seeded samples. Normalized band-area ratios for collagen content of single cell-seeded samples gradually decreased during a 21-day culture period, whereas collagen content of the microaggregate-seeded samples significantly increased during this period. Moreover, a fibre-optic Raman set-up allowed for the collection of Raman spectra from multiple pores inside scaffolds in parallel. These fibre-optic measurements could give a representative average of the ECM Raman signal present in tissue-engineered constructs. Results in this study provide proof-of-principle that Raman microspectroscopy is a promising non-invasive tool to monitor ECM production and remodelling in three-dimensional porous cartilage tissue-engineered constructs.

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