Secretion of Hematopoietic Niche Signal Molecules under Conditions of Osteogenic Differentiation of Multipotent Mesenchymal Stromal Cells Induced By Relief Calcium Phosphate Coating

L. S. Litvinova, V. V. Shupletsova, K. A. Yurova, O. G. Khaziakhmatova, N. M. Todosenko, V. V. Malashchenko, E. O. Shunkin, E. S. Melashchenko, M. Yu Khlusova, E. G. Komarova, V. V. Chebodaeva, Yu P. Sharkeev, P. A. Ivanov, I. A. Khlusov

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Abstract—: Using a multiplex kit the secretion of a number of cytokines, chemokines, and growth factors has been investigated in vitro in a culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) under conditions of their osteogenic differentiation caused by 14-day contact with a calcium phosphate (CP) surface of different roughness. Bilateral X-ray amorphous CP coatings were prepared on the samples of commercially pure titanium in the anodal regime using a microarc method. The electrolyte consisted of aqueous orthophosphoric acid (20 wt %), calcium carbonate (9 wt %), and synthetic hydroxyapatite nanopowder (6 wt %, particle diameter of 10–30 nm with single agglomerates up to 100 nm). hAMMSCs isolated from lipoaspirate were co-cultured after 4 passages with the CP-coated samples at a final concentration of 1.5 × 105 viable karyocytes per 1.5 mL of standard nutrition medium (without osteogenic stimulators) for 14 days (determination of the [CD45,34,14,20], CD73, CD90, and CD105 cell immunophenotype; analysis of secretory activity) and 21 days (alizarin red S cell culture staining) with medium replacement every 3–4 days. Under conditions of in vitro contact with rough CP coating hAMMSCs differentiated into osteoblasts synthesizing the mineralized bone matrix; this was accompanied by a 2−3-fold increase in the proportion of [CD45,34,14,20]+ hemopoietic cells. The following humoral factors of hemopoietic niches acted as the signal molecules escalating in vitro the hemopoietic base in 14 days of differentiating three-dimensional culture of hAMMSCs: leukemia inhibitory factor (LIF) and stem cell factor (SCF) cytokines in the case of the mean index of CP roughness Ra = 2.4–2.6 µm or stromal derived factor-1 (SDF-1α, CXCL12 chemokine) in the case of Ra = 3.1–4.4 µm.

Original languageEnglish
Pages (from-to)341-348
Number of pages8
JournalBiochemistry (Moscow) Supplement Series B: Biomedical Chemistry
Volume13
Issue number4
DOIs
Publication statusPublished - 1 Oct 2019

Keywords

  • in vitro
  • microarc coating
  • roughness index R
  • stem cell factor
  • stem cell growth factor
  • stromal derived factor-1

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Clinical Biochemistry

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