Abstract

Cell therapies present a feasible option for the treatment of degenerated cartilaginous and intervertebral disc (IVD) tissues. Microenvironments of these tissues are specific and often differ from the microenvironment of cells that, could be potentially used for therapy, e.g. human adipose-derived stem cells (hASC). To ensure safe and efficient implantation of hASC, it is important to evaluate how microenvironmental conditions at the site of implantation affect the implanted cells. This study has demonstrated that cartilaginous tissue-specific osmolarities ranging from 400–600 mOsm/L affected hASC in a dose- and time-dependent fashion in comparison to 300 mOsm/L. Increased osmolarities resulted in transient (nuclear DNA and actin reorganisation) and non-transient, long-term morphological changes (vesicle formation, increase in cell area, and culture morphology), as well as reduced proliferation in monolayer cultures. Increased osmolarities diminished acid proteoglycan production and compactness of chondrogenically induced pellet cultures, indicating decreased chondrogenic potential. Viability of hASC was strongly dependent on the type of culture, with hASC in monolayer culture being more tolerant to increased osmolarity compared to hASC in suspension, alginate-agarose hydrogel, and pellet cultures, thus emphasizing the importance of choosing relevant in vitro conditions according to the specifics of clinical application.

Highlights

  • Degeneration of cartilaginous tissues is a serious health problem, which affects a large percentage of the worldwide population

  • Human adipose-derived stem cells cultured in 400 mOsm/L, 500 mOsm/L, and 600 mOsm/L remained attached and had a comparable viability to that of cells cultured in 300 mOsm/L up to 4 w (Fig 1A–1D)

  • Human adipose-derived stem cells are the subject of many studies focusing on their chondrogenic and anti-inflammatory [43, 44, 45] potential in the context of clinical applications for cartilaginous tissue repair [13, 17, 18, 43, 46, 47, 48, 49]

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Summary

Introduction

Degeneration of cartilaginous tissues is a serious health problem, which affects a large percentage of the worldwide population. Low back pain affects up to 85% of people during their lives and represents a high social, healthcare, and economic burden [1, 2]. Cell therapies represent a possible approach for the treatment of intervertebral disc (IVD) and cartilage degeneration [3, 4, 5]. Human adipose-derived stem cells (hASC) have gained significant interest as a cell source due to their accessibility, limited donor site damage, high proliferation rate, and differentiation potential [5, 6, 7, 8, 9, 10, 11, 12]. PLOS ONE | DOI:10.1371/journal.pone.0163870 October 5, 2016

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