An in vitro analysis of the effect of hyperosmolarity on the chondrogenic potential of human articular cartilage derived chondroprogenitors

Abstract

Purpose: Chondroprogenitors display promise for articular cartilage regeneration. It is imperative to standardize culture conditions, to further enhance chondrogenicity and reduce tendency for hypertrophy. Cartilage matrix provides a unique hyperosmolar microenvironment that enables native cells to resist compressive stress. However, commonly used culture media have osmolarities relatively hypoosmotic when compared to in-vivo conditions. Previous reports involving chondrocytes demonstrated enhanced chondrogenic potential secondary to utilization of hyperosmolar culture conditions. The study aimed to assess the effect of hyperosmolarity (either mimicking normal joint conditions or short-term hyperosmotic stress) on chondroprogenitor phenotype.

Materials and methods: Fibronectin adhesion assay derived human articular chondroprogenitors (n = 3) were divided into 3 groups: a) Control: cells grown in standard culture conditions (320 mOsm/L), b) Test A: cells grown in hyperosmolar media mimicking joint conditions (409 mOsm/L) and c) Test B: cells exposed to short-term hyperosmotic stress (504 mOsm/L) for 24 h, prior to assessment. Evaluation parameters included population doubling, cell size, surface marker expression, mRNA expression (markers of chondrogenesis, dedifferentiation and hypertrophy) and multilineage potential.

Results: Subjecting these cells to increased osmolarity in culture did not demonstrably favor chondrogenesis (control vs Test A: comparable COL2A1) while hyperosmotic stress further increased the tendency for hypertrophy and terminal differentiation (high COL1A1 and low COL2A1, P = 0.006). Additionally, growth kinetics, surface marker expression and multilineage potential were comparable across groups.

Conclusion: Chondroprogenitors displayed sensitivity to increase in osmolarity as chondrogenic phenotype did not improve, while hypertrophic propensity was heightened, although further analysis of culture and phenotypic parameters will aid in optimizing chondroprogenitor use in cartilage regeneration.

Keywords: Cartilage regeneration; Chondrogenesis; Chondroprogenitors; Hyperosmolarity; Hypertrophy.