Passage and reversal effects on gene expression of bovine meniscal fibrochondrocytes

Abstract

The knee meniscus contains a mixed population of cells that exhibit fibroblastic as well as chondrocytic characteristics. Tissue engineering studies and future therapies for the meniscus require a large population of cells that are seeded on scaffolds. To achieve this, monolayer expansion is often used as a technique to increase cell number. However, the phenotype of these cells may be significantly different from that of the primary population. The objective of this study was to investigate changes in meniscal fibrochondrocytes at the gene expression level over four passages using quantitative real-time reverse transcriptase polymerase chain reaction. Cells from the inner two-thirds of bovine medial menisci were used. Four extracellular matrix (ECM) molecules, commonly found in the meniscus, were investigated, namely collagen I, collagen II, aggrecan and cartilage oligomeric matrix protein (COMP). In addition, primary and passaged meniscus fibrochondrocytes were placed on surfaces coated with collagen I or aggrecan protein to investigate whether any gene expression changes resulting from passage could be reversed. Collagen I expression was found to increase with the number of passages, whereas collagen II and COMP expression decreased. Collagen I and aggrecan surface coatings were shown to downregulate and upregulate collagen I and COMP expression levels, respectively, in passaged cells. However, decreases in collagen II expression could not be reversed by either protein coating. These results indicate that although monolayer expansion results in significant changes in gene expression in meniscal fibrochondrocytes, protein coatings may be used to regain the primary cell expression of several ECM molecules.

Porosity and Cell Preseeding Influence Electrospun Scaffold Maturation and Meniscus IntegrationIn Vitro

“…41 Despite this, culture-expanded cells are likely phenotypically distinct from the native meniscus cells due to changes during monolayer expansion. 42 Thus, it is unclear if the growth differences seen between tissue-free control scaffolds and scaffold-meniscus constructs are due to changes in cells themselves with passaging, or to the presence of the native tissue. Similarly, meniscus cells for preseeding or preculture were derived from all portions of the meniscus and combined; a more detailed analysis should be conducted using region-specific cell populations.…”

Biological Augmentation and Tissue Engineering Approaches in Meniscus Surgery

“…Given the relative scarcity of these cells in the tissue, monolayer expansion is also required, which can result in downregulation of genes involved in matrix production. 76 Allogenic and xenogeneic cell sources have shown promise in animal models, and although they may overcome the discrepancy in supply and demand, they are fraught with translational issues. 77,78 MSCs, in contrast, have been successfully isolated from adipose tissue, muscle, and bone marrow and subsequently used in soft tissue engineering.…”

Citocompatibilidade de blendas de poli(p-dioxanona)/ poli(hidroxi butirato) (PPD/PHB) para aplicações em engenharia de tecido cartilaginoso

“…Estas moléculas apresentam importantes funções biomecânicas [57] . O colágeno do tipo I é um dos principais colágenos encontrados nesta matriz [58] . Naumann et al [59] caracterizaram os componentes da matriz extracelular do menisco de coelhos e observaram marcação positiva para elastina e colágeno VI na proximidade das células do menisco.…”