Abstract
BackgroundBone marrow stromal cells (BMSC) have promise in cartilage tissue engineering, but for their potential to be fully realised, the propensity to undergo hypertrophy must be mitigated. The literature contains diverging reports on the effect of parathyroid hormone (PTH) on BMSC differentiation. Cartilage tissue models can be heterogeneous, confounding efforts to improve media formulations.MethodsHerein, we use a novel microwell platform (the Microwell-mesh) to manufacture hundreds of small-diameter homogeneous micro-pellets and use this high-resolution assay to quantify the influence of constant or intermittent PTH(1–34) medium supplementation on BMSC chondrogenesis and hypertrophy. Micro-pellets were manufactured from 5000 BMSC each and cultured in standard chondrogenic media supplemented with (1) no PTH, (2) intermittent PTH, or (3) constant PTH.ResultsRelative to control chondrogenic cultures, BMSC micro-pellets exposed to intermittent PTH had reduced hypertrophic gene expression following 1 week of culture, but this was accompanied by a loss in chondrogenesis by the second week of culture. Constant PTH treatment was detrimental to chondrogenic culture.ConclusionsThis study provides further clarity on the role of PTH on chondrogenic differentiation in vitro and suggests that while PTH may mitigate BMSC hypertrophy, it does so at the expense of chondrogenesis.
Highlights
Bone marrow stromal cells (BMSC) have promise in cartilage tissue engineering, but for their potential to be fully realised, the propensity to undergo hypertrophy must be mitigated
In chondrogenic human BMSC pellet cultures, intermittent parathyroid hormone (PTH) treatment appeared to restore chondrogenic differentiation that was lost in cultures treated with constant PTH, leading to reduced Indian hedgehog (IHH) expression and alkaline phosphatase (ALP) activity [23]. These findings suggest that a PTH treatment regimen may be important in controlling the differentiation of BMSC
The tubes were centrifuged for 30 min at 400×g, and interface cells were collected, washed, and resuspended in low glucose Dulbecco’s modified Eagle’s medium (LG-DMEM; Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific), 10 ng/mL fibroblast growth factor-1 (FGF-1; PeproTech), and 100 U/mL penicillin/streptomycin (PenStrep; Thermo Fisher Scientific)
Summary
Bone marrow stromal cells (BMSC) have promise in cartilage tissue engineering, but for their potential to be fully realised, the propensity to undergo hypertrophy must be mitigated. Articular cartilage is an avascular, aneural tissue with limited regenerative potential. PTH significantly increased aggrecan levels in chondrocytes isolated from day 17 and 18 embryos, but no increase was observed in chondrocytes harvested from embryos at day 20 or 21 of gestation [16]. These observations suggest that the stage of chondrogenesis may be important for PTH signalling outcomes
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
