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Cell death in human articular chondrocyte: a morpho-functional study in micromass model

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Abstract

Chondrocyte death and loss of extracellular matrix are the central features in articular cartilage degeneration during osteoarthritis pathogenesis. Cartilage diseases and, in particular, osteoarthritis are widely correlated to apoptosis but, chondrocytes undergoing apoptosis “in vivo” more often display peculiar features that correspond to a distinct process of programmed cell death termed “chondroptosis”. Programmed cell death of primary human chondrocyte has been here investigated in micromasses, a tridimensional culture model, that represents a convenient means for studying chondrocyte biology. Cell death has been induced by different physical or chemical apoptotic agents, such as UVB radiation, hyperthermia and staurosporine delivered at both 1 and 3 weeks maturation. Conventional electron microscopy was used to analyse morphological changes. Occurrence of DNA fragmentation and caspase involvement were also investigated. At Transmission Electron Microscopy, control cells appear rounding or slightly elongated with plurilobated nucleus and diffusely dispersed chromatin. Typically UVB radiation and staurosporine induce chromatin apoptotic features, while hyperthermia triggers the “chondroptotic” phenotype. A weak TUNEL positivity appears in control, correlated to the well known cell death patterns occurring along cartilage differentiation. UVB radiation produces a strong positivity, mostly localized at the micromass periphery. After hyperthermia a higher number of fluorescent nuclei appears, in particular at 3 weeks. Staurosporine evidences a diffuse, but reduced, positivity. Therefore, DNA fragmentation is a common pattern in dying chondrocytes, both in apoptotic and “chondroptotic” cells. Moreover, all triggers induce caspase pathway activation, even if to a different extent, suggesting a fundamental role of apoptotic features, in chondrocyte cell death.

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Acknowledgments

This research was supported by Urbino University and MIUR PRIN 2009 (Michela Battistelli, Sara Salucci and Elisabetta Falcieri); CARISBO Foundation of Bologna, (Eleonora Olivotto, Stefania Pagani and Andrea Facchini); FIRB-MIUR, Grant RBAP10KCNS (Andrea Facchini, Annalisa Facchini, Flavio Flamigni and Rosa Maria Borzì); Fondi cinque per mille (Ministero della Salute,) (Andrea Facchini and Rosa Maria Borzì); POS-FESR 2007–2013, Emilia Romagna Region (EO).

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Authors and Affiliations

  1. DiSTeVA, Campus Scientifico Enrico Mattei, Università degli Studi di Urbino Carlo Bo, Via Ca’ le Suore 2, 61029, Urbino, PU, Italy

    M. Battistelli, S. Salucci & E. Falcieri

  2. Laboratorio di Immunoreumatologia E Rigenerazione Tissutale/Laboratorio RAMSES Istituti Ortopedici Rizzoli, Bologna, 40100, Italy

    E. Olivotto, M. Minguzzi, S. Guidotti, S. Pagani, R. M. Borzì & A. Facchini

  3. Dipartimento di Scienze Biomediche E Neuromotorie, Università degli Studi di Bologna, Bologna, 40100, Italy

    A. Facchini & F. Flamigni

  4. Dipartimento di Scienze Mediche E Chirurgiche, Università degli Studi di Bologna, Bologna, 40100, Italy

    A. Facchini, M. Minguzzi, S. Guidotti & A. Facchini

  5. IGM, CNR, Istituti Ortopedici Rizzoli, Bologna, 40100, Italy

    E. Falcieri

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  1. M. Battistelli
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  2. S. Salucci
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Correspondence to M. Battistelli.

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Battistelli, M., Salucci, S., Olivotto, E. et al. Cell death in human articular chondrocyte: a morpho-functional study in micromass model. Apoptosis 19, 1471–1483 (2014). https://doi.org/10.1007/s10495-014-1017-9

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