Abstract
IntroductionIn the current study, we investigated the effect of low magnitude, low frequency (LMLF) mechanical vibrations on the osteogenic differentiation potential of human adipose derived mesenchymal stem cells (hASC), taken from elderly patients.MethodsDuring 21 days in osteogenic culture medium, cells were periodically exposed to three different frequencies (25, 35 and 45 Hz) of continuous sinusoidal oscillation, using a vibration generator. We measured cell proliferation, cell morphology, calcium and phosphorus deposition using Almar Blue assay, fluorescence microscopy, scanning electron microscopy, and a EDX detector, respectively. Osteogenic differentiation was measured by assessing protein and mRNA levels. Osteogenesis was confirmed by detection of specific markers with alkaline phosphatase and enzyme-linked immunosorbent assays for: bone morphogenetic protein 2 (BMP-2), osteocalcin (OCL) and osteopontin (OPN).Results We found that 25 Hz vibrations had the greatest impact on hASC morphology, ultrastructure, and proliferation. We observed the formation of osteocyte- and hydroxyapatite-like structures, an increased quantity of calcium and phosphorus deposits, and increased differentiation in the stimulated groups.Conclusions Our findings suggest that LMLF vibrations could be used to enhance cell-based therapies for treatment of bone deficits, particularly in elderly patients, where the need is greatest.
Highlights
Bone tissue engineering is a rapidly developing field of regenerative medicine
Immunophenotypic characterization of human adipose derived mesenchymal stem cells (hASC) obtained from the patient samples confirmed the presence of mesenchymal markers (CD29, CD44, CD73, CD90) and excluded hematopoietic origin (CD45, CD34) of the obtained cells (Fig. 2)
In cells exposed to 25 Hz vibration, the proliferation factor (PF) at day 18 was higher, and their population doubling time (PDT) was significantly (p < 0.01) lower as compared to controls (Fig. 4)
Summary
Bone tissue engineering is a rapidly developing field of regenerative medicine This rapid expansion is in part driven by the ever-growing increase in the number of patients who stand to benefit from these treatments. These include patients who have sustained bone injuries and tissue loss due to trauma, cancer, osteonecrosis, osteoarthritis, and the ever growing population of elderly patients in whom the bone’s natural regenerative capacities are reduced, resulting in altered healing properties.[30]. While many treatments are available, the gold standard in reconstructive orthopedics continues to be vascularized or non-vascularized autologous bone grafts.
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