Arthritic periosteal tissue from joint replacement surgery: a novel, autologous source of stem cells

Abstract

The overarching aim of this study is to assess the feasibility of using periosteal tissue from the femoral neck of arthritic hip joints, usually discarded in the normal course of hip replacement surgery, as an autologous source of stem cells. In addition, the study aims to characterize intrinsic differences between periosteum-derived cell (PDC) populations, isolated via either enzymatic digestion or a migration assay, including their proliferative capacity, surface marker expression, and multipotency, relative to commercially available human bone marrow-derived stromal cells (BMSCs) cultured under identical conditions. Commercial BMSCs and PDCs were characterized in vitro, using a growth assay, flow cytometry, as well as assay of Oil Red O, alizarin red, and Safranin O/Fast Green staining after respective culture in adipo-, osteo-, and chondrogenic media. Based on these outcome measures, PDCs exhibited proliferation rate, morphology, surface receptor expression, and multipotency similar to those of BMSCs. No significant correlation was observed between outcome measures and donor age or diagnosis (osteoarthritis [OA] and rheumatoid arthritis [RA], respectively), a profound finding given recent rheumatological studies indicating that OA and RA share not only common biomarkers and molecular mechanisms but also common pathophysiology, ultimately resulting in the need for joint replacement. Furthermore, PDCs isolated via enzymatic digestion and migration assay showed subtle differences in surface marker expression but otherwise no significant differences in proliferation or multipotency; the observed differences in surface marker expression may indicate potential effects of isolation method on the population of cells isolated and/or the behavior of the respective isolated cell populations. This study demonstrates, for the first time to our knowledge, the feasibility of using arthritic tissue resected during hip replacement as a source of autologous stem cells. In sum, periosteum tissue that is resected with the femoral neck in replacing the hip represents an unprecedented and, to date, unstudied source of stem cells from OA and RA patients. Follow-up studies will determine the degree to which this new, autologous source of stem cells can be banked for future use.

Keywords: Arthritis; Clinical translation; Differentiation; Osteoarthritis; Periosteum; Regenerative medicine; Rheumatoid arthritis; Stem cell.

Figure 1.

Cell morphology and growth curves. (A–C): Representative phase contrast (×10) microscopic images of BMSCs (A), dPDCs (B), and mPDCs (C). Cells exhibited similar morphologies and sizes. Scale bar = 100 μm. (D): Growth curves of cells were quantified for 15 days. All cell types showed similar growth rates (slope) (n = 4). Error bars indicate 95% confidence intervals. Abbreviations: BMSC, bone marrow-derived stromal cell; dPDC, enzymatically digested periosteum-derived cells; mPDC, migrated periosteum-derived cell.

Figure 2.

Oil Red O staining after culture in adipogenic medium. (A–F): Control BMSCs (A), dPDCs (B), and mPDCs (C) and induced BMSCs (D), dPDCs (E), and mPDCs (F) stained with Oil Red O. Scale bar = 100 μm. (G): Oil Red O absorbance for control and induced cells. Six replicates of each donor are included to show consistency and distribution. Asterisks (∗) indicate difference from the control (p < .05) (n = 4). Error bars indicate 95% confidence intervals. Abbreviations: BMSC, bone marrow-derived stromal cell; dPDC, enzymatically digested periosteum-derived cells; mPDC, migrated periosteum-derived cell.

Figure 3.

Alizarin red staining after culture in osteogenic medium. (A–F): Control BMSCs (A), dPDCs (B), and mPDCs (C) and induced BMSCs (D), dPDCs (E), and mPDCs (F), stained with alizarin red. Each well represents a single donor: top left, age 30; top right, age 48; bottom left, age 58; and bottom right, age 72. (G): Alizarin red absorbance for control and induced cells. Six replicates of each donor are included to show consistency and distribution (n = 4). Error bars indicate 95% confidence intervals. (H): Alizarin red absorbance for different donors. Plus sign (+) indicates increase from control, and asterisk (∗) indicates decrease from control (p < .05) (n = 6). Error bars indicate 95% confidence intervals. Abbreviations: BMSC, bone marrow-derived stromal cell; dPDC, enzymatically digested periosteum-derived cells; mPDC, migrated periosteum-derived cell.

Figure 4.

Safranin O/Fast Green staining after culture in chondrogenic medium. (A–F): Pellets of control (A–C) and induced (D-F) BMSCs, dPDCs, and mPDCs, stained for GAG production with Safranin O/Fast Green. Scale bars = 250 and 50 μm. (G): Cross-sectional pellet area of control and induced cells. Three replicates of each donor are included to show consistency and distribution. Asterisks (∗) indicate difference from the control (p < .05) (n = 3). Error bars indicate 95% confidence intervals. Abbreviations: BMSC, bone marrow-derived stromal cell; Chondro, chondrogenic medium; dPDC, enzymatically digested periosteum-derived cells; mPDC, migrated periosteum-derived cell.