Umbilical cord stem cells and pulsed electromagnetic fields: potential tools for tendon repair?
ESSKA Academy. Marmotti A. May 11, 2018; 218052
Dr. Antonio Marmotti
Dr. Antonio Marmotti
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Objectives: The use of umbilical cord mesenchymal stem cells(UC-MSC) and pulsed electromagnetic fields to improve tendon repair is still investigational.The combination of PEMF, and UC-MSC supplementation may have clinical relevance for improving tendon cell-based tissue engineering.The aim of the study is to evaluate in vitro the effect of low-frequency PEMF on tenogenic differentiation of MSC isolated from human UC-MSC

Methods: 15 Fresh UC samples from women with healthy pregnancies were retrieved at the end of caesarean deliveries. The UC samples were manually minced without any enzymatic digestion into very small fragments and cultivated in an MSC expansion medium, then UC tissue was removed and adherent cells were allowed to expand. At day 28, the adherent cells were collected and replated until confluence (passage 1, P1).
Immunophenotypic characterization was performed.
For tendon differentiation the medium was: DMEM, 10% FCS, 50 U/ml penicillin, 50 lg/ml streptomycin, 2 mM L-glutamine and 5 ng/ml b-FGF.
the following cell cultures have been set up:
1) UC-MSCs with MSC differentiation medium + exposure to PEMF
2) UC-MSCs with MSC differentiation medium without exposure to PEMF
3) UC-MSCs with MSC-grown base medium (without b-FGF) without exposure to PEMF
intensity of magnetic field=1.5 mT,frequency=75Hz
The UC-MSCs were subjected to PEMF for 4 h/day (PEMF1) or 8 h/day (PEMF2) .
At 7, 14, 21 days of differentiation, the apoptosis was analyzed by flow cytometry, using annexin V/propidium iodide. At the same time points immunofluorescence analysis was performed to evaluate the expression of collagen type I and scleraxis (markers of tendon differentiation) and PCNA (proliferative marker).

Results: At P1, we obtained a mean value of 0.78 (SD 0.28) x 10E6 cells/gr of UC.
Cells were positive for CD73, CD90, CD105, CD44, CD29 and HLA-I, and negative for CD34 and HLA-class II.
The UC-MSCs in the presence of FGF-2 and PEMF showed greater production of collagen type I and scleraxis than controls without PEMF and without PEMF and FGF-2 (p<0,05): the expression of this markers increased from 7 to 21 days of culture, both in PEMF1 than PEMF2. PEMF2 showed a better expression than PEMF1 of the specific tendon differentiation markers. The expression of PCNA, in the presence of FGF-2 and PEMF, was significantly lower (p<0,05) than controls without PEMF and without PEMF and FGF-2. The exposure to PEMF did not influence cell viability

Conclusions: The use of PEMF provides a mechanical stimulus that seems to enhance tendon differentiation of UC-MSC. Clinical relevance is that
i) the newly described tenogenic effects of PEMF may represent a positive therapeutic tool to be used during tendon repair procedure
ii) the combined use of PEMF and UC-MSC may represent a future perspective, with a strong in-vitro rationale shown by this work, to ameliorate cell-based tissue engineering of specific joints, i.e. the shoulder enthesis, that may benefits of both chondrogenic and tenogenic improvement

tendon differentiation umbilical cord mesenchymal stem cells pulsed electromagnetic fields
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