Co-culture of chondrons and mesenchymal stromal cells reduces the loss of collagen VI and improves extracellular matrix production

Owida, Hamza Abu; Ruiz, T De Las Heras; Dhillon, Anisa; Yang, Ying; Kuiper, Nicola J.

doi:10.1007/s00418-017-1602-4

Cited by 21 publications

(24 citation statements)

References 36 publications

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“…Recent works have shown that co-culture of human chondrons with MSCs produce more cartilage ECM than chondrocytes with MSCs [49,50]. In this study, we found that co-culture of chondrons with chondrocytes, particularly at a ratio of 1:1, significantly elevated the levels of AGG and Col-2 expression and GAG production in vitro, suggesting that co-culture of them at a good ration may promote chondrogenesis.…”

Section: Discussionsupporting

confidence: 55%

Combination of chondrocytes with chondrons improves Proteoglycan and Collagen-2 production to promote the repairs of defective knee cartilage in rabbits

Duan

Zhao

Ren

et al. 2020

Preprint

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Background: Chondrons are composed of chondrocytes and the surrounding pericellular matrix (PCM) and function to enhance chondrocyte-mediated cartilage tissue engineering. This study aimed at investigating the potential effect of combined chondrocytes with chondrons on the production of proteoglycan and collagen-II (Col-2) and the repair of defective knee cartilage in rabbits.Methods: Chondrocytes and chondrons were isolated from the knee cartilage of rabbits, and cultured alone or co-cultured for varying periods in vitro. Their morphology was characterized by histology and immunohistochemistry. The levels of aggrecan (AGG), Col-2 and glycosaminoglycan (GAG) expression were quantified by qRT-PCR, Alcian Blue-based precipitation and ELISA. The effect of combined chondrocytes with chondrons in alginate spheres on the repair of defective knee cartilage was examined in rabbits.Results: The isolated chondrocytes and chondrons displayed unique morphology and began to proliferate on day 3 and 6 post culture, respectively, accompanied by completely degenerated PCM on day 6 post culture. Evidently, chondrocytes had stronger proliferation capacity than chondrons. Longitudinal analyses indicated that culture of chondrons, but not chondrocytes, increased AGG mRNA transcripts and GAG levels with time and Col-2 mRNA transcripts only on day 3 post culture. Compared with chondrocytes or chondrons alone, co-culture of chondrocytes and chondrons significantly up-regulated AGG and Col-2 expression and GAG production, particularly at a ratio of 1:1. Implantation with chondrocytes and chondrons at 1:1 significantly promoted the repair of defective knee cartilage in rabbits, accompanied by reduced the Wakiteni scores with time. Conclusion: Combined chondrons with chondrocytes promoted the production of proteoglycan and Col-2 and the repair of defective knee cartilage in rabbits.

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Section: Discussionsupporting

confidence: 55%

Combination of chondrocytes with chondrons improves Proteoglycan and Collagen-2 production to promote the repairs of defective knee cartilage in rabbits

Duan

Zhao

Ren

et al. 2020

Preprint

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“…Four separate isolations were performed, each using one humerus. Chondrocyte isolations were performed with a minor modification of our previously described protocol [19]. Briefly, diced cartilage was sequentially digested with 0.1% (w/v) proteinase K for 1 h and then 0.3% (w/v) collagenase type IA for 3 h. Chondrocytes from the digestion supernatant were strained through a 70 mm cell sieve and washed in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% (v/v) fetal calf serum (FCS).…”

Section: Isolation Of Chondrocytesmentioning

confidence: 99%

Induction of zonal-specific cellular morphology and matrix synthesis for biomimetic cartilage regeneration using hybrid scaffolds

Owida

Yang

Chen

et al. 2018

J. R. Soc. Interface.

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Cartilage is anisotropic in nature and organized into distinct zones. Our goal was to develop zonal-specific three-dimensional hybrid scaffolds which could induce the generation of zonal-specific cellular morphology and extracellular matrix (ECM) composition. The superficial and middle zones comprised two layers of hyaluronic acid (HA) hydrogel which enveloped specifically orientated or randomly arranged polylactic acid nanofibre meshes. The deep zone comprised a HA hydrogel with multiple vertical channels. Primary bovine chondrocytes were seeded into the individual zonal scaffolds, cultured for 14 days and then the ECM was analysed. The aligned nanofibre mesh used in the superficial zone induced an elongated cell morphology, lower glycosaminoglycan (GAG) and collagen II production, and higher cell proliferation and collagen I production than the cells in the middle zone scaffold. Within the middle zone scaffold, which comprised a randomly orientated nanofibre mesh, the cells were clustered and expressed more collagen II. The deep zone scaffold induced the highest GAG production, the lowest cell proliferation and the lowest collagen I expression of the three zones. Assembling the three zones and stabilizing the arrangement with a HA hydrogel generated aligned, randomly aggregated and columnar cells in the superficial, middle and deep zones. This study presents a method to induce zonal-specific chondrocyte morphology and ECM production.

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“…Images were whitebalanced using Photoshop Elements (Adobe). To semi-quantify the toluidine blue staining intensity, four central images of each pellet were randomly selected and the integrated density was determined using ImageJ Software (NIH) according to the published methodology (Jensen, 2013;Owida et al, 2017;Prasad and Prabhu, 2012).…”

Section: Hypertrophic Markers Alk1/acvrl1mentioning

confidence: 99%

Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

Sykes¹,

Kuiper²,

Richardson³

et al. 2018

eCM

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High hopes have been pinned on regenerative medicine strategies in order to prevent the progression of cartilage damage to osteoarthritis, particularly by autologous chondrocyte implantation (ACI). The loss of chondrocyte phenotype during in vitro monolayer expansion, a necessary step to obtain sufficient cell numbers, may be a key limitation in ACI. In this study, it was determined whether a shorter monolayer expansion approach could improve chondrogenic differentiation. The effects of two supplement types, foetal bovine serum (FBS) and Stemulate™ (a commercial source of human platelet lysate), on the expansion and re-differentiation potential of human chondrocytes, isolated from five individuals, were compared. Chondrocytes were expanded with 10 % FBS or 10 % Stemulate™. Pellets were cultured for 28 d in chondrogenic differentiation medium and assessed for the presence of cartilage matrix molecules and genes associated with chondrogenicity. Stemulate™ significantly enhanced the proliferation rate [average population doubling times: FBS, 25.07 ± 6.98 d (standard error of the mean, SEM) vs. Stemulate™, 13.10 ± 2.57 d (SEM)]. Sulphated glycosaminoglycans (sGAG), total collagen and qRT-PCR analyses of cartilage genes showed that FBS-expanded chondrocytes demonstrated significantly better chondrogenic capacity than Stemulate™-expanded chondrocytes. Histologically, FBS-expanded chondrocyte pellets appeared to be more stable, with a more intense staining for toluidine blue, indicating a greater chondrogenic capacity. Although Stemulate™ positively influenced chondrocyte proliferation, it had a negative effect on chondrogenic differentiation potential. This suggested that, in the treatment of cartilage defects, Stemulate™ might not be the ideal supplement for expanding chondrocytes (which maintained a chondrocyte phenotype) and, hence, for cell therapies (including ACI).

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Co-culture of chondrons and mesenchymal stromal cells reduces the loss of collagen VI and improves extracellular matrix production

Cited by 21 publications

References 36 publications

Combination of chondrocytes with chondrons improves Proteoglycan and Collagen-2 production to promote the repairs of defective knee cartilage in rabbits

Combination of chondrocytes with chondrons improves Proteoglycan and Collagen-2 production to promote the repairs of defective knee cartilage in rabbits

Induction of zonal-specific cellular morphology and matrix synthesis for biomimetic cartilage regeneration using hybrid scaffolds

Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

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