Identification of the skeletal progenitor cells forming osteophytes in osteoarthritis (original) (raw)
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Arthritis Research & Therapy, 2011
Introduction: Mesenchymal stem cells (MSC) are highly attractive for use in cartilage regeneration. To date, MSC are usually recruited from subchondral bone marrow using microfracture. Recent data suggest that isolated cells from adult human articular cartilage, which express the combination of the cell-surface markers CD105 and CD166, are multi-potent mesenchymal progenitor cells (MPC) with characteristics similar to MSC. MPC within the cartilage matrix, the target of tissue regeneration, may provide the basis for in situ regeneration of focal cartilage defects. However, there is only limited information concerning the presence/abundance of CD105 + /CD166 + MPC in human articular cartilage. The present study therefore assessed the relative percentage and particularly the zonal distribution of cartilage MPC using the markers CD105/CD166. Methods: Specimens of human osteoarthritic (OA; n = 11) and normal (n = 3) cartilage were used for either cell isolation or immunohistochemistry. Due to low numbers, isolated cells were expanded for 2 weeks and then analyzed by flow cytometry (FACS) or immunofluorescence in chamber slides for the expression of CD105 and CD166. Following immunomagnetic separation of CD166 + / -OA cells, multi-lineage differentiation assays were performed. Also, the zonal distribution of CD166 + cells within the matrix of OA and normal cartilage was analyzed by immunohistochemistry. Results: FACS analysis showed that 16.7 ± 2.1% (mean ± SEM) of OA and 15.3 ± 2.3 of normal chondrocytes (n.s.) were CD105 + /CD166 + and thus carried the established MPC marker combination. Similarly, 13.2% ± 0.9% and 11.7 ± 2.1 of CD105 + /CD166 + cells, respectively, were identified by immunofluorescence in adherent OA and normal chondrocytes. The CD166 + enriched OA cells showed a stronger induction of the chondrogenic phenotype in differentiation assays than the CD166 + depleted cell population, underlining the chondrogenic potential of the MPC. Strikingly, CD166 + cells in OA and normal articular cartilage sections (22.1 ± 1.7% and 23.6% ± 1.4%, respectively; n.s.) were almost exclusively located in the superficial and middle zone. Conclusions: The present results underline the suitability of CD166 as a biomarker to identify and, in particular, localize and/or enrich resident MPC with a high chondrogenic potential in human articular cartilage. The percentage of MPC in both OA and normal cartilage is substantially higher than previously reported, suggesting a yet unexplored reserve capacity for regeneration.
Journal of Orthopaedic Research®, 2017
Cell-based therapies development for the treatment of osteoarthritis (OA) requires an understanding of the disease progression and attributes of the cells resident in cartilage. This study focused on quantitative assessment of the concentration and biological potential of stem and progenitor cells resident in different zones of cartilage displaying macroscopic Outerbridge grade 1-2 OA, and their correlation with OA progression based on established histologic scoring system. Lateral femoral condyles were collected from 15 patients with idiopathic OA and varus knees undergoing total knee arthroplasty. Superficial(C sp , top $ 500 mm) and deep cartilage(C dp) was separated. Chondrogenic Connective Tissue Progenitors (CTP-C) were assayed by standardized Colony-Forming-Unit assay using automated image analysis (Colonyze TM) based on ASTM standard F-2944-12. Cell concentration (cells/mg) was significantly greater in C sp (median: 7,000; range: 3,440-17,600) than C dp (median: 5,340; range: 3,393-9,660), p ¼ 0.039. Prevalence (CTPs/million cells) was not different between C sp (median: 1,274; range: 0-3,898) and C dp (median:1,365; range:0-6,330), p ¼ 0.42. In vitro performance of CTP-C progeny varied widely within and between patients, manifest by variation in colony size and morphology. Mean histopathological Mankin score was 4.7 (SD ¼ 1.2), representing mild to moderate OA. Tidemark breach by blood vessels was associated with lower C sp cell concentration (p ¼ 0.02). Matrix degradation was associated with lower C dp cell and CTP-C concentration (p ¼ 0.015 and p ¼ 0.095, respectively), independent of articular surface changes. These findings suggest that the initiation of OA may occur in either superficial or deep zones. The pathological changes affect CTP-Cs in C sp and C dp cartilage zones differently. The heterogeneity among the available CTP-Cs in C sp and C dp suggests performance-based selection to optimize cell-sourcing strategies for therapy.
Scientific reports, 2018
Adult human articular cartilage harbors a population of CD166+ mesenchymal stem cell-like progenitors that become more numerous during osteoarthritis (OA). While their role is not well understood, here we report that they are indeed part of cellular clusters formed in OA cartilage, which is a pathological hallmark of this disease. We hypothesize that these cells, termed OA mesenchymal stem cells (OA-MSCs), contribute to OA pathogenesis. To test this hypothesis, we generated and characterized multiple clonally derived stable/immortalized human OA-MSC cell lines, which exhibited the following properties. Firstly, two mesenchymal stem cell populations exist in human OA cartilage. While both populations are multi-potent, one preferentially undergoes chondrogenesis while the other exhibits higher osteogenesis potential. Secondly, both OA-MSCs exhibit significantly higher expression of hypertrophic OA cartilage markers COL10A1 and RUNX2, compared to OA chondrocytes. Induction of chondroge...
Biotechnology Progress, 2012
Tissue engineering efforts for the fabrication of cartilage substitutes head toward applicability in osteoarthritis (OA). Progenitor cells can be harvested from the osteoarthritic joint itself, resembling multipotent mesenchymal stromal cells (MSC). Our objective was to analyze MSC characteristics of those cells in respect to the OA-related damage of their harvest site. OA cartilage was obtained from six patients during alloarthroplastic knee surgery, sample grading was done according to Outerbridge's classification. Upon enzymatic dissociation, primary chondrocytes were expanded in two-dimensional monolayer culture. At distinct cell passages, the process of dedifferentiation was phenotypically monitored; cell surface expression of classical MSC markers was analyzed by flow cytometry. Cells were subjected to chondrogenesis and osteogenesis after their fourth passage. At third passage, 95% of cells became positive for cluster of differentiation (CD)105 and further subclassification revealed that the majority of them were positive for both CD73 and CD90. CD105 þ CD73 þ CD90 þ phenotype meets thus the minimal surface antigen criteria for MSC definition. More than one-third of dedifferentiated chondrocytes displayed a coexpression of CD9 þ CD166 þ CD90 þ and to a lesser extent CD105 þ CD73 þ CD44 þ , irrespective of the stage of the original cartilage degradation. Finally, we could successfully demonstrate a redifferentiation of these progenitors into sulfated glycosaminoglycan producing cells. The basic level of alkaline phosphatase activity could not be enhanced upon osteogenic differentiation. In conclusion, chondrogenic progenitors derived from OA cartilages with low or high Outerbridge's grade can be seen as a potential cellular source for cartilage replacement. V
Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature, 2018
Adult human articular cartilage harbors a population of CD166+ mesenchymal stem cell-like progenitors that become more numerous during osteoarthritis (OA). While their role is not well understood, here we report that they are indeed part of cellular clusters formed in OA cartilage, which is a pathological hallmark of this disease. We hypothesize that these cells, termed OA mesenchymal stem cells (OA-MSCs), contribute to OA pathogenesis. To test this hypothesis, we generated and characterized multiple clonally derived stable/immortalized human OA-MSC cell lines, which exhibited the following properties. Firstly, two mesenchymal stem cell populations exist in human OA cartilage. While both populations are multi-potent, one preferentially undergoes chondrogenesis while the other exhibits higher osteogenesis potential. Secondly, both OA-MSCs exhibit significantly higher expression of hypertrophic OA cartilage markers COL10A1 and RUNX2, compared to OA chondrocytes. Induction of chondrogenesis in OA-MSCs further stimulated COL10A1 expression and MMP-13 release, suggesting that they contribute to OA phenotypes. Finally, knocking down RUNX2 is insufficient to inhibit COL10A1 in OA-MSCs and also requires simultaneous knockdown of NOTCH1 thereby suggesting altered gene regulation in OA stem cells in comparison to chondrocytes. Overall, our findings suggest that OA-MSCs may drive pathogenesis of cartilage degeneration and should therefore be a novel cell target for OA therapy.
Journal of Advanced Biotechnology and Experimental Therapeutics, 2021
ABSTRACT: Osteoarthritis (OA) is a leading pathological condition resulting in the degeneration and destruction of articular cartilage. The presence of inherent mesenchymal progenitor cells (MPCs) within the articular cartilage has led to explore the possible reparative mechanisms to regenerate and restore the functional and mechanical properties of hyaline cartilage. The present in vitro study was aimed to identify and characterize the MPCs derived from OA cartilage. MPCs derived from the explant culture of OA cartilage were analyzed in terms of cellular and biological properties, and multilineage differentiation abilities. Upon cell surface marker analysis, MPCs were CD73+, CD90+, CD166+, CD146-, CD34-, CD45-, and HLA-DR-, whose expression defines stemness and chondroprogenitor status. MPCs exhibited a higher proliferative index and limited or no senescence activity till later passages. Trilineage differentiation towards osteogenesis, adipogenesis, and chondrogenesis was observed with cytochemical staining and also by mRNA expression of lineagespecific markers by RT-qPCR. The results showed that OA cartilage harbors a viable pool of MPCs with greater chondrogenic potential. These cell niches could serve as a superior cell source for cartilage regeneration due to their committed progeny and hence could prevent heterotypic cartilage formation.
Journal of Bone and Joint Surgery, 2018
Background: Current decisions on cellular therapies for osteoarthritis are based primarily on clinical experience or on assumptions about preferred cell sourcing. They have not been informed by rigorous standardized measurements of the chondrogenic connective-tissue progenitors (CTP-Cs) or their intrinsic diversity of chondrogenic potential. The goal of this study was to quantitatively define the CTP-Cs resident in cartilage of different grades of osteoarthritis and to compare their concentration, prevalence, and biological potential. Methods: Twenty-three patients who had varus malalignment of the knee and were scheduled to undergo elective total knee arthroplasty for idiopathic osteoarthritis and who had grade 1-2 osteoarthritis on the lateral femoral condyle and grade 3-4 osteoarthritis on the medial femoral condyle were recruited for study of the cartilage removed during surgery. CTP-Cs were assayed by a standardized colony-forming-unit assay using automated image-analysis software based on ASTM standard test method F2944-12. Results: Cell concentration was significantly greater (p < 0.001) in grade 3-4 cartilage than in grade 1-2 cartilage. The prevalence of CTP-Cs varied widely, but it trended lower in grade 3-4 cartilage than in grade 1-2 samples (p = 0.078). The biological performance of CTP-Cs from grade 1-2 and grade 3-4 cartilage was comparable. Increased cell concentration was a significant predictor of decreased CTP-C prevalence (p = 0.002). Conclusions: Although grade 3-4 cartilage showed fewer CTP-Cs than grade 1-2 cartilage, the range of biological performance was comparable, which suggests that either may be used as a source for potent CTP-Cs. However, the biological reason for the heterogeneity of CTP-Cs in cartilage and the biological implications of that heterogeneity are not well understood and require further study. Clinical Relevance: In order to improve the efficacy of cartilage cell therapy procedures, it is key to characterize the quality and quantity of the cells and progenitors being administered. Additionally, understanding the heterogeneity in order to select appropriate subsets of populations will improve the rigor of decisions concerning cell sourcing and targeting for pharmacological and cellular therapies.
Journal of tissue engineering, 2011
The expression of type X collagen (COL X), a late-stage chondrocyte hypertrophy marker in human mesenchymal stem cells (MSCs) from osteoarthritis (OA) patients poses a major setback to current cartilage and intervertebral disc tissue engineering efforts. However, it is not yet clear whether COL X is expressed by all human bone marrow stem cells or if it is related to age, gender, site, disease status, or drug therapy. In the current study, we report that COL X expression is upregulated in MSCs from rabbits in a surgical instability model of OA (anterior cruciate ligament transection (ACLT)) when compared to control rabbit MSCs. Thus COL X expression in OA is a common phenomenon that is due to the disease process itself and not to other environmental factors. It is, therefore, critical to understand MSC phenotype in OA patients, as these cells are essential clinically for biological repair of cartilage lesions using autologous stem cells.