Bone Marrow Mesenchymal Stem Cells in Patients with Beta Thalassemia Major: Molecular Analysis with Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy Study as a Novel Method

Abstract

Bone marrow mesenchymal stem cells (BM-MSCs) are the main cellular components of the bone marrow, providing a supportive cellular microenvironment to maintain healthy hematopoiesis. β-thalassemia major (β-TM) is characterized by anemia that is caused by a genetic defect in hemoglobin synthesis and results in ineffective erythropoiesis (IE). The alterations in the microenvironment in thalassemic bone marrow during IE can cause changes in BM-MSCs. This study aimed to investigate global structural and compositional changes in BM-MSCs in β-TM that may provide a basis in understanding interactions of hematopoietic stem cells (HSCs)-MSCs in such a pathological bone marrow microenvironment. Following characterization of morphological, immunophenotypical, and differentiation properties, the changes in healthy and thalassemic BM-MSCs before and after bone marrow transplantation (BMT) were examined by attenuated total reflection-Fourier transform infrared (ATR-FTIR). The significant increase in lipid, protein, glycogen, and nucleic acid contents in thalassemic BM-MSCs with respect to healthy BM-MSCs was attributed to enhanced cell proliferation and BM activity during IE. The significant decreases in the content of mentioned macromolecules in post-transplant group BM-MSCs versus pre-transplant BM-MSCs was interpreted as restoring effect of BMT therapy on IE and defective BM microenvironment. These alterations were also supported by ELISA results of erythropoietin (EPO) and growth differentiation factor (GDF15) in bone marrow plasma samples as a reflection of IE and by MTT proliferation assay on BM-MSCs. Based on these changes, sampling groups were discriminated by cluster analysis. These results provide information for the studies that concentrate on interactions between HSCs-MSCs in bone marrow.

The Phenotype and Functional Activity of Mesenchymal Stromal Cells in Pediatric Patients with Non-Malignant Hematological Diseases

“…MSCs of patients with NMHD were able to equally differentiate into osteoblasts and chondrocytes as MSCs of healthy donors. Similar results for MSCs of patients with thalassemia [29,30] and SCD [32,34] were reported. Interestingly, we observed that MSCs of all four tested patients with thalassemia differentiated less well into adipocytes than MSCs of other NMHD and control group.…”

“…In addition, MSCs of patients with thalassemia and SCD demonstrated an equivalent proliferation potential as MSCs of healthy donors as reported by several groups [32,34,35]. Only one group reported that thalassemic MSCs proliferate more than MSCs of healthy donors [30], in contrast to Crippa et al [31] who observed an opposite phenomenon. Flow cytometry analysis showed a significantly lower expression of CD146 antigen, as a more primitive cell marker, on MSCs of patients with TM, SCD and SCN compared to MSCs of healthy donors.…”

“…However, MSCs of patients with non-malignant hematological diseases have been less well studied. In contrast to MSCs generated from patients with aplastic anemia (AA), which have been thoroughly investigated [22][23][24][25][26][27][28], MSCs from patients with thalassemia and SCD are poorly characterized [29][30][31][32][33][34][35], whereas MSCs generated from patients with congenital neutropenia are not studied at all. In the current study, we show for the first time that bone marrow of thalassemia patients contain a significantly higher number of mononuclear cells (BM-MNCs) than the bone marrow of healthy donors.…”

“…According to Yoon et al [29], who reported data from one patient, there is no difference in the quality of MSCs as morphology, phenotype, proliferation, trilineage differentiation potential and immunosuppression in mixed lymphocyte reaction (MLR) compared to control. Aksoy et al [30] reported that MSCs from thalassemia patients exerted also a higher proliferation potential than MSCs of healthy donors. Recently, Crippa et al [31] confirmed a reduced frequency of primitive MSCs in β-thalassemia patients caused by increased reactive oxygen species (ROS) production in vitro which impaired MSC stemness properties.…”

Application of Fourier transform infrared spectroscopy to biomolecular profiling of cultured fibroblast cells from Gaucher disease patients: A preliminary investigation

“…Samples were measured after hav-ing been dried on the crystal, similar to the method applied in the literature. 14,15 Briefly, fibroblast cell samples were mixed gently prior to the measurements to obtain homogeneity. 2.5 µL of cell suspension in PBS were spotted onto the ZnSe ATR crystal.…”