Redifferentiation of aged human articular chondrocytes by combining bone morphogenetic protein-2 and melanoma inhibitory activity protein in 3D-culture

Payr, Stephan; Tichy, Brigitte; Atteneder, Clemens; Michel, Mary Ellen; Tiefenboeck, Thomas M.; Lang, Nikolaus; Nuernberger, Sylvia; Hajdu, Stefan; Balmayor, Elizabeth R.; Marlovits, Stefan; Albrecht, Christian

doi:10.1371/journal.pone.0179729

Cited by 6 publications

(5 citation statements)

References 35 publications

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“…ACAN, COL2A1, and GAG are known to be the most components of cartilage ECM. In vitro, ACAN, COL2A1, and GAG biosynthesis support chondrocyte redifferentiation [41]. Therefore, changes in ACAN, COL2A1, and GAG reflect the anabolism of cartilage ECM.…”

Section: Discussionmentioning

confidence: 99%

^pNNS-Conjugated Chitosan Mediated IGF-1 and miR-140 Overexpression in Articular Chondrocytes Improves Cartilage Repair

Zhao

Zhang

Jia

et al. 2019

BioMed Research International

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The aim of the present study was to investigate the effects of phosphorylatable nucleus localization signal linked nucleic kinase substrate short peptide (NNSp)-conjugated chitosan (NNSp-CS) mediated miR-140 and IGF-1 in both rabbit chondrocytes and cartilage defects model. NNSp-CS was combined with pBudCE4.1-IGF-1, pBudCE4.1-miR-140, and negative control pBudCE4.1 to form pDNA/NNSp-CS complexes. Then these complexes were transfected into chondrocytes or injected intra-articularly into the knee joints. High levels of IGF-1 and miR-140 expression were detected both in vitro and in vivo. Compared with pBudCE4.1 group, in vitro, the transgenic groups significantly promoted chondrocyte proliferation, increased glycosaminoglycan (GAG) synthesis, and ACAN, COL2A1, and TIMP-1 levels, and reduced the levels of nitric oxide (NO), MMP-13, and ADAMTS-5. In vivo, the exogenous genes enhanced COL2A1, ACAN, and TIMP-1 expression in cartilage and reduced cartilage Mankin score and the contents of NO, IL-1β, TNF-α, and GAG contents in synovial fluid of rabbits, MMP-13, ADAMTS-5, COL1A2, and COL10A1 levels in cartilage. Double gene combination showed better results than single gene. This study indicate that NNSp-CS is a better gene delivery vehicle in gene therapy for cartilage defects and that miR-140 combination IGF-1 transfection has better biologic effects on cartilage defects.

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Section: Discussionmentioning

confidence: 99%

^pNNS-Conjugated Chitosan Mediated IGF-1 and miR-140 Overexpression in Articular Chondrocytes Improves Cartilage Repair

Zhao

Zhang

Jia

et al. 2019

BioMed Research International

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“…25 Although the use of BMP2 has been related to hypertrophy of chondrocytes and calcification of cartilage ECM associated with increased expression of COL10 mRNA and protein, [26][27][28] Payr et al have argued that hypertrophy and expression of osteogenic markers after BMP2 stimulation might be more relevant when using MSCs than human articular chondrocytes. 29 Still, the role played by BMP2 in the CDMB deserves further investigation. With respect to IGF1, a study on the redifferentiation capacity of human chondrocytes in pellet cultures has shown that TGFβ1 alone had positive effects on expression of ECM components under normoxia, whereas a positive effect of IGF1 in combination with TGFβ1 was only seen under hypoxic conditions.…”

Section: Discussionmentioning

confidence: 98%

“…have argued that hypertrophy and expression of osteogenic markers after BMP2 stimulation might be more relevant when using MSCs than human articular chondrocytes. 29 Still, the role played by BMP2 in the CDMB deserves further investigation.…”

Section: Discussionmentioning

confidence: 98%

Scaffold-Free Engineering of Human Cartilage Implants

et al. 2021

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Objective Despite new strategies in tissue engineering, cartilage repair remains a major challenge. Our aim is to treat patients with focal lesions of articular cartilage with autologous hyaline cartilage implants using a scaffold-free approach. In this article, we describe experiments to optimize production of scaffold-free cartilage discs. Design Articular chondrocytes were expanded in vitro, seeded in transwell inserts and redifferentiated using established chondrogenic components. Experimental variables included testing 2 different expansion media, adding bone morphogenetic protein 2 (BMP2), insulin-like growth factor 1 (IGF1), growth/differentiation factor 5 (GDF5), or fibroblast growth factor 18 (FGF18) to the differentiation medium and allowing the disc to float freely in large wells. Cartilage discs were analyzed by weight and thickness, real-time RT-qPCR (reverse transcriptase qualitative polymerase chain reaction), fluorescence immunostaining, transmission electron microscopy, second harmonic generation imaging, and measurement of Young’s modulus. Results Addition of BMP2 to the chondrogenic differentiation medium (CDM) was essential for stable disc formation, while IGF1, GDF5, and FGF18 were redundant. Allowing discs to float freely in CDM on a moving platform increased disc thickness compared with discs kept continuously in transwell inserts. Discs cultured for 6 weeks reached a thickness of almost 2 mm and Young’s modulus of >200 kPa. There was abundant type II collagen. Collagen fibrils were 25 nm thick, with a tendency to be organized perpendicular to the disc surface. Conclusion Scaffold-free engineering using BMP2 and providing free movement in CDM produced firm, elastic cartilage discs with abundant type II collagen. This approach may potentially be used in clinical trials.

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“…In vitro , chondrocyte redifferentiation was supported by COL2A1, ACAN, and GAG biosynthesis. 45 Changes in the COL2A1, ACAN, and GAG levels therefore reflect the anabolic level of the cartilage ECM. 46 In this study, compared with the control group, the groups with IL-1Ra and miR-140 overexpression each individually presented with increased GAG accumulation in the cell supernatants and COL2A1 and ACAN expression in the chondrocyte, and the joint IL-1Ra and miR-140 treatment significantly enhanced these effects.…”

Section: Discussionmentioning

confidence: 99%

Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model

Zhao

Wang

Jia

et al. 2019

Bone & Joint Research

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Objectives Previously, we reported the improved transfection efficiency of a plasmid DNA-chitosan (pDNA-CS) complex using a phosphorylatable nuclear localization signal-linked nucleic kinase substrate short peptide (pNNS) conjugated to chitosan (pNNS-CS). This study investigated the effects of pNNS-CS-mediated miR-140 and interleukin-1 receptor antagonist protein (IL-1Ra) gene transfection both in rabbit chondrocytes and a cartilage defect model. Methods The pBudCE4.1-miR-140, pBudCE4.1-IL-1Ra, and negative control pBudCE4.1 plasmids were constructed and combined with pNNS-CS to form pDNA/pNNS-CS complexes. These complexes were transfected into chondrocytes or injected into the knee joint cavity. Results High IL-1Ra and miR-140 expression levels were detected both in vitro and in vivo. In vitro , compared with the pBudCE4.1 group, the transgenic group presented with significantly increased chondrocyte proliferation and glycosaminoglycan (GAG) synthesis, as well as increased collagen type II alpha 1 chain (COL2A1), aggrecan (ACAN), and TIMP metallopeptidase inhibitor 1 (TIMP-1) levels. Nitric oxide (NO) synthesis was reduced, as were a disintegrin and metalloproteinase with thrombospondin type 1 motif 5 (ADAMTS-5) and matrix metalloproteinase (MMP)-13 levels. In vivo , the exogenous genes reduced the synovial fluid GAG and NO concentrations and the ADAMTS-5 and MMP-13 levels in cartilage. In contrast, COL2A1, ACAN, and TIMP-1 levels were increased, and the cartilage Mankin score was decreased in the transgenic group compared with the pBudCE4.1 group. Double gene combination produced greater efficacies than each single gene, both in vitro and in vivo . Conclusion This study suggests that pNNS-CS is a good candidate for treating cartilage defects via gene therapy, and that IL-1Ra in combination with miR-140 produces promising biological effects on cartilage defects. Cite this article : R. Zhao, S. Wang, L. Jia, Q. Li, J. Qiao, X. Peng. Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model. Bone Joint Res 2019;8:165–178. DOI: 10.1302/2046-3758.83.BJR-2018-0222.R1.

show abstract

Redifferentiation of aged human articular chondrocytes by combining bone morphogenetic protein-2 and melanoma inhibitory activity protein in 3D-culture

Cited by 6 publications

References 35 publications

^pNNS-Conjugated Chitosan Mediated IGF-1 and miR-140 Overexpression in Articular Chondrocytes Improves Cartilage Repair

^pNNS-Conjugated Chitosan Mediated IGF-1 and miR-140 Overexpression in Articular Chondrocytes Improves Cartilage Repair

Scaffold-Free Engineering of Human Cartilage Implants

Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model

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