Engineering cartilage tissue is challenging, mainly because chondrocytes lose their differentiated phenotype when cultured in monolayer. The aim of this study was to analyse the influence of 3D-culture conditions on the redifferentiation of chondrocytes, devoting special attention to BMPs. Dedifferentiated chondrocytes were seeded onto two different scaffolds (Bio-Gide and HYAFF-11) and were then cultured for 38 days. Every week, samples were taken for gene expression analysis and immunohistochemistry. In both scaffolds an increasing differentiation was observed caused by an increase in Col2 and a reduction in Col1 expression. The various BMPs were regulated, albeit differently by the changing culture conditions. While GDF-5 and BMP-4 expression increased in the monolayer culture in comparison with native cartilage and decreased again in the 3D culture, the BMP-2 and BMP-6 expression decreased dramatically in the monolayer as well as in the 3D culture. BMP-7 was not detectable in any probe. Scaffold cultivation appears to stimulate the induction of redifferentiation, but is not sufficient to induce expression of BMP-2 -6 or -7. Since, in comparison to cartilage development, there is a lack of surrounding signal centres, external stimuli seem to be required to obtain complete redifferentiation. Our data indicate that a combination of BMP-2, -6 and -7 may be promising for this purpose.
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Biotechnol Bioeng
December 2024
Department of Joint Surgery, The First Affiliated Hospital, Guangzhou Medical University, Guangzhou, China.
Chondrocytes maintain the balance of the extracellular matrix by synthesizing glycoproteins, collagen, proteoglycans and hyaluronic acid. Chondrocyte dedifferentiation refers to a process in which chondrocytes lose their mature differentiated phenotype and transform into a fibroblast-like morphology with fewer differentiated stages and inferior function under external stimulation. The important mechanism of homeostasis loss in osteoarthritis (OA) is a change in the chondrocyte phenotype.
View Article and Find Full Text PDFEur J Cell Biol
June 2024
Department of Biological Sciences, University of Delaware, USA; Department of Biomedical Engineering, University of Delaware, USA. Electronic address:
Tissue Eng Regen Med
July 2024
Department of Facial Plastic and Reconstructive Surgery, Eye and ENT Hospital, Fudan University, Shanghai, 200031, China.
Background: Tissue engineering is increasingly viewed as a promising avenue for functional cartilage reconstruction. However, chondrocyte dedifferentiation during in vitro culture remains an obstacle for clinical translation of tissue engineered cartilage. Re-differentiated induction have been employed to induce dedifferentiated chondrocytes back to their original phenotype.
View Article and Find Full Text PDFACS Omega
May 2024
School of Pharmacy and Biomedical Sciences, University of Portsmouth, St Michael's Building, White Swan Road, Portsmouth PO1 2DT, U.K.
Focal cartilage defects are a prevalent knee problem affecting people of all ages. Articular cartilage (AC) possesses limited healing potential, and osteochondral defects can lead to pain and long-term complications such as osteoarthritis. Autologous chondrocyte implantation (ACI) has been a successful surgical approach for repairing osteochondral defects over the past two decades.
View Article and Find Full Text PDFBiotechnol Lett
June 2024
Department of Joint Surgery, The Key Laboratory of Advanced Interdisciplinary Studies Center, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510120, Guangdong, China.
Objectives: We genetically modified dedifferentiated chondrocytes (DCs) using lentiviral vectors and adenoviral vectors encoding TGF-β3 (referred to as transgenic groups below) and encapsulated these DCs in the microcavitary hydrogel and investigated the combinational effect on redifferentiation of the genetically manipulated DCs.
Results: The Cell Counting Kit-8 data indicated that both transgenic groups exhibited significantly higher cell viability in the first week but inferior cell viability in the subsequent timepoints compared with those of the control group. Real-time polymerase chain reaction and western blot analysis results demonstrated that both transgenic groups had a better effect on redifferentiation to some extent, as evidenced by higher expression levels of chondrogenic genes, suggesting the validity of combination with transgenic DCs and the microcavitary hydrogel on redifferentiation.
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