Bone marrow aspirate concentrate quality is affected by age and harvest site

Carola Cavallo; Angelo Boffa; Laura de Girolamo; Giulia Merli; Elizaveta Kon; Luca Cattini; Emma Santo; Brunella Grigolo; Giuseppe Filardo

doi:10.1007/s00167-022-07153-6

Bone marrow aspirate concentrate quality is affected by age and harvest site

Knee Surg Sports Traumatol Arthrosc. 2023 Jun;31(6):2140-2151. doi: 10.1007/s00167-022-07153-6. Epub 2022 Sep 26.

Authors

Carola Cavallo¹, Angelo Boffa², Laura de Girolamo³, Giulia Merli⁴, Elizaveta Kon^{5

6}, Luca Cattini⁷, Emma Santo¹, Brunella Grigolo¹, Giuseppe Filardo⁴

Affiliations

¹ Laboratorio RAMSES, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy.
² Applied and Translational Research (ATR) Center, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy. angeloboffa@libero.it.
³ Laboratorio di Biotecnologie Applicate all'Ortopedia, IRCCS Istituto Ortopedico Galeazzi, Milan, Italy.
⁴ Applied and Translational Research (ATR) Center, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy.
⁵ Humanitas Clinical and Research Center - IRCCS, Rozzano, Italy.
⁶ Humanitas University, Department of Biomedical Sciences, Pieve Emanuele, Italy.
⁷ Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy.

Abstract

Purpose: To compare the number and properties of bone marrow stromal cells (BMSCs) collected from bone marrow aspirate concentrate (BMAC) obtained from different harvest sites and from patients of different ages.

Methods: BMAC was obtained from two groups of patients based on age (n = 10 per group): 19.0 ± 2.7 years for the younger and 56.8 ± 12.5 for the older group. In the latter, BMAC was obtained from both iliac crest and proximal tibia for a donor-matched analysis. Mononucleated cell count and CFU-F assay were performed, together with phenotype characterization of BMSCs from iliac crest and proximal tibia, the study of chondrogenic and osteogenic differentiation capacity, histological staining and spectrophotometric quantification, and the analysis of mRNAs expression.

Results: Cells derived from iliac crest and proximal tibia showed the same phenotypic pattern at flow cytometry, as well as similar chondrogenic and osteogenic potential. However, a significantly higher number of mononuclear cells per ml was observed in younger patients (3.8 ± 1.8 × 10⁷) compared to older patients (1.2 ± 0.8 × 10⁷) (p < 0.0005). The latter yield, obtained from the iliac crest, was significantly higher than resulting from the BMAC harvested from the proximal tibia in the same group of patients (0.3 ± 0.2 × 10⁷, p < 0.0005). This result was confirmed by the CFU-F analysis at day 10 (15.9 ± 19.4 vs 0.6 ± 1.0, p = 0.001) and day-20 (21.7 ± 23.0 vs 2.9 ± 4.2, p = 0.006).

Conclusion: Harvest site and age can affect the quality of BMAC. BMSCs obtained from iliac crest and proximal tibia present comparable mesenchymal markers expression as well as osteogenic and chondrogenic differentiation potential, but iliac crest BMAC presents a four times higher number of mononucleated cells with significantly higher clonogenic capacity compared to the tibia. BMAC of younger patients also had a three-time higher number of mononucleated cells. The identification of BMAC characteristics could help to optimize its preparation and to identify the most suitable indications for this orthobiologic treatment in the clinical practice.

Keywords: Age; BMAC; Bone marrow aspirate concentrate; Harvest site; Iliac crest; MSCs; Tibia.

MeSH terms

Bone Marrow*
Cell Differentiation
Mesenchymal Stem Cells*
Osteogenesis
Stem Cells / metabolism