articular chondrocytes
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Author(s):  
Simon L. Wuest ◽  
Geraldine Cerretti ◽  
Jennifer Louise Wadsworth ◽  
Cindy Follonier ◽  
Karin F. Rattenbacher-Kiser ◽  
...  

2021 ◽  
Author(s):  
Ni Zeng ◽  
Zhi-Peng Yan ◽  
Tao Liao ◽  
Jie-Ting Li ◽  
Yan Chen ◽  
...  

Abstract Objective: To investigate the interaction of infrapatellar fat pad/cartilage and related mechanisms in knee osteoarthritis (OA) using the metabolomics method.Method: Fat-conditioned media (FCM) of the infrapatellar fat pad from patients with OA were used to treat human OA chondrocytes. The extracellular metabolites of human OA chondrocytes were detected by nontargeted metabolic footprint analysis based on liquid chromatography and mass spectrometry (LC-MS). Then, the different metabolites were found, and the main metabolic pathways were explored, combined with bioinformatics methods.Results: After treatment with FCM for 48 h, the proliferation of human OA chondrocytes was slowed down, indicating that FCM had a certain inhibitory effect on the proliferation of human OA chondrocytes (P = 0.023). On the pattern diagram of principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA), after FCM treatment, the data sample areas were obviously separated, indicating that FCM can significantly affect the metabolic footprint of human OA chondrocytes. Through metabonomic identification, 131 different metabolites were screened after FCM treatment compared with before treatment. For 4 pathways in total, significantly different activity levels were discovered in pairwise comparisons: alanine, aspartate, and glutamate metabolism; citrate cycle (TCA cycle); arginine and proline metabolism; and phenylalanine metabolism.Conclusion: The infrapatellar fat pad aggravates OA chondrocyte injury and is involved in OA by disturbing the chondrocyte TCA cycle, amino acid metabolism, and glutamine metabolism, among others.


2021 ◽  
Vol 129 ◽  
pp. 110774
Author(s):  
Junpei Hatakeyama ◽  
Masato Nomura ◽  
Yoshio Wakimoto ◽  
Shota Inoue ◽  
Changxin Li ◽  
...  

2021 ◽  
Author(s):  
Kosei Nagata ◽  
Hironori Hojo ◽  
Song Ho Chang ◽  
Hiroyuki Okada ◽  
Fumiko Yano ◽  
...  

Abstract The Runt-related transcription factor (Runx) family plays various roles in the homeostasis of cartilage. Here, we examined the role of Runx2 and Runx3 for osteoarthritis (OA) development in vivo and in vitro. Runx3 knockout mice accelerated OA by surgical induction, accompanied with decreased expression of lubricin and aggrecan. Meanwhile, Runx2 conditional knockout mice showed biphasic phenotypes; OA was inhibited by hetero-knockout accompanied with decreased matrix metallopeptidase 13 (Mmp13) expression, but accelerated in homo-knockout of Runx2 accompanied with reduction of type II collagen (Col2a1) expression. Comprehensive transcriptional analyses revealed lubricin and aggrecan as transcriptional target genes of Runx3, and indicated that Runx2 sustained Col2a1 expression through an intron 6 enhancer when Sox9 was decreased. Intra-articular administration of Runx3 adenovirus ameliorated development of surgically induced OA. Runx3 protects adult articular cartilage through extracellular matrix protein production under the normal condition, while Runx2 exerts both catabolic and anabolic effects under the inflammatory condition.


Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2343
Author(s):  
Aitthiphon Chongchai ◽  
Sajee Waramit ◽  
Tunchanok Wongwichai ◽  
Jirawan Kampangtip ◽  
Thanyaluck Phitak ◽  
...  

Osteoarthritis (OA) is a degenerative joint disease characterized by progressive deterioration and loss of articular cartilage. There is currently no treatment to reverse the onset of OA. Thus, we developed a targeted delivery strategy to transfer genes into primary human chondrocytes as a proof-of-concept study. We displayed a chondrocyte-affinity peptide (CAP) on the pIII minor coat protein of the M13 filamentous bacteriophage (phage)-based particle carrying a mammalian transgene cassette under cytomegalovirus CMV promoter and inverted terminal repeats (ITRs) cis elements of adeno-associated virus serotype 2 (AAV-2). Primary human articular chondrocytes (HACs) were used as an in vitro model, and the selectivity and binding properties of the CAP ligand in relation to the pathogenic conditions of HACs were characterized. We found that the CAP ligand is highly selective toward pathogenic HACs. Furthermore, the stability, cytotoxicity, and gene delivery efficacy of the CAP-displaying phage (CAP.Phage) were evaluated. We found that the phage particle is stable under a wide range of temperatures and pH values, while showing no cytotoxicity to HACs. Importantly, the CAP.Phage particle, carrying a secreted luciferase (Lucia) reporter gene, efficiently and selectively delivered transgene expression to HACs. In summary, it was found that the CAP ligand preferably binds to pathogenic chondrocytes, and the CAP.Phage particle successfully targets and delivers transgene to HACs.


2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Marta R. Casanova ◽  
Hugo Osório ◽  
Rui L. Reis ◽  
Albino Martins ◽  
Nuno M. Neves

AbstractExtracellular vesicles (EVs) are being increasingly studied owing to its regenerative potential, namely EVs derived from human bone marrow mesenchymal stem cells (hBM-MSCs). Those can be used for controlling inflammation, repairing injury, and enhancing tissue regeneration. Differently, the potential of EVs derived from human articular chondrocytes (hACs) to promote cartilage regeneration has not been thoroughly investigated. This work aims to develop an EVs immobilization system capable of selectively bind EVs present in conditioned medium obtained from cultures of hACs or hBM-MSC. For that, an anti-CD63 antibody was immobilized at the surface of an activated and functionalized electrospun nanofibrous mesh. The chondrogenic potential of bound EVs was further assessed by culturing hBM-MSCs during 28 days under basal conditions. EVs derived from hACs cultured under differentiation medium or from chondrogenically committed hBM-MSCs induced a chondrogenic phenotype characterized by marked induction of SOX9, COMP, Aggrecan and Collagen type II, and matrix glycosaminoglycans synthesis. Indeed, both EVs immobilization systems outperformed the currently used chondroinductive strategies. These data show that naturally secreted EVs can guide the chondrogenic commitment of hBM-MSCs in the absence of any other chemical or genetic chondrogenic inductors based in medium supplementation.


2021 ◽  
Vol 22 (21) ◽  
pp. 11503
Author(s):  
Lukas B. Moser ◽  
Christoph Bauer ◽  
Vivek Jeyakumar ◽  
Eugenia-Paulina Niculescu-Morzsa ◽  
Stefan Nehrer

The current study aimed to investigate the cytotoxicity of co-administrating local anesthetics (LA) with glucocorticoids (GC) and hyaluronic acid (HA) in vitro. Human articular cartilage was obtained from five patients undergoing total knee arthroplasty. Chondrocytes were isolated, expanded, and seeded in 24-well plates for experimental testing. LA (lidocaine, bupivacaine, ropivacaine) were administered separately and co-administered with the following substances: GC, HA, and GC/HA. Viability was confirmed by microscopic images, flow cytometry, metabolic activity, and live/dead assay. The addition of HA and GC/HA resulted in enhanced attachment and branched appearance of the chondrocytes compared to LA and LA/GC. Metabolic activity was better in all LA co-administered with HA and GC/HA than with GC and only LA. Flow cytometry revealed the lowest cell viability in lidocaine and the highest cell viability in ropivacaine. This finding was also confirmed by live/dead assay. In conclusion, HA supports the effect of GC and reduces chondrotoxic effects of LA in vitro. Thereby, the co-administration of HA to LA and GC offers an alternative less chondrotoxic approach for treating patients with symptomatic osteoarthritis of the knee.


Author(s):  
Jade Tassey ◽  
Arijita Sarkar ◽  
Ben Van Handel ◽  
Jinxiu Lu ◽  
Siyoung Lee ◽  
...  

Cartilage tissue is comprised of extracellular matrix and chondrocytes, a cell type with very low cellular turnover in adults, providing limited capacity for regeneration. However, in development a significant number of chondrocytes actively proliferate and remodel the surrounding matrix. Uncoupling the microenvironmental influences that determine the balance between clonogenic potential and terminal differentiation of these cells is essential for the development of novel approaches for cartilage regeneration. Unfortunately, most of the existing methods are not applicable for the analysis of functional properties of chondrocytes at a single cell resolution. Here we demonstrate that a novel 3D culture method provides a long-term and permissive in vitro niche that selects for highly clonogenic, colony-forming chondrocytes which maintain cartilage-specific matrix production, thus recapitulating the in vivo niche. As a proof of concept, clonogenicity of Sox9IRES–EGFP mouse chondrocytes is almost exclusively found in the highest GFP+ fraction known to be enriched for chondrocyte progenitor cells. Although clonogenic chondrocytes are very rare in adult cartilage, we have optimized this system to support large, single cell-derived chondrogenic organoids with complex zonal architecture and robust chondrogenic phenotype from adult pig and human articular chondrocytes. Moreover, we have demonstrated that growth trajectory and matrix biosynthesis in these organoids respond to a pro-inflammatory environment. This culture method offers a robust, defined and controllable system that can be further used to interrogate the effects of various microenvironmental signals on chondrocytes, providing a high throughput platform to assess genetic and environmental factors in development and disease.


Cartilage ◽  
2021 ◽  
pp. 194760352110354
Author(s):  
Yunliang Lei ◽  
Jiabin Peng ◽  
Zhu Dai ◽  
Ying Liao ◽  
Quanhui Liu ◽  
...  

Objective This study was undertaken to elucidate the mechanism of improved chondrocyte migration after juvenile articular cartilage fragmentation. Design In vitro organ culture with rabbit cartilage fragments and cell culture with rabbit chondrocytes were performed. In part A, minced juvenile cartilage fragments (~0.5 × 0.5 × 0.5 mm) from rabbits, planted in gelatin sponge and fibrin glue, were cultured for 2, 4, or 6 weeks in vitro and compared with the cartilage chunks (~4 × 4 × 1 mm) and membrane type 1 matrix metalloprotease (MT1-MMP) inhibitor groups. Chondrocyte outgrowth was evaluated on histology and confocal laser scanning microscopy. MT1-MMP expression was compared between the cartilage fragment group and the cartilage chunks group. In part B, articular chondrocytes were harvested from juvenile rabbits, MT1-MMP was transfected into the cells, and cell migration was evaluated using the Transwell and wound healing tests. Results The histology and confocal microscopy results revealed that cell accumulation occurred at the edge of cartilage fragments, and outgrowth was better in the cartilage fragment group than those in the cartilage chunks group. Similar results were observed for MT1-MMP expression. After MT1-MMP inhibition, cells did not accumulate at the edge of the cartilage fragments, and chondrocyte outgrowth did not occur. Furthermore, overexpression of MT1-MMP enhanced the migration of articular chondrocytes. Conclusions Juvenile articular cartilage fragmentation improved chondrocyte migration by upregulating MT1-MMP.


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