Expressions of pathologic markers in PRP based chondrogenic differentiation of human adipose derived stem cells
Graphical abstract
Section snippets
Background
In the recent decade, PRP has been widely used as a therapeutic for OA patients with pain killing effects, although its overall effects depend on the stage of the disease, or in the other words on the micro-environmental conditions. The application of activated Platelet-Rich Plasma (PRP), with its vast range of cytokines and growth factors has achieved a considerable attention for chondrogenic differentiation in tissue engineering fields.
PRP is rich in growth factors such as TGF-β, Insulin-like
Adipose stem cells isolation, expansion and characterization
Liposuctioned samples were collected from healthy patients after obtaining their written informed consent according to the ethics of Tehran University of Medical Sciences. For ADSCs isolation briefly, the liposuctioned sample was washed several times to remove the blood cells, 150 μg/ml collagenase I (Sigma) was added and the sample was incubated for 60 min at 37 °C while on shaker at 250 rpm. Collagenase was removed by centrifugation at 1200 rpm for 5 min. The pellet was then incubated with RBC
Chondrogenic marker expression in PRP-based differentiation
Cartilage specific markers such as SOX9, aggrecan and Collagen type II genes were expressed in all groups, and as presented PRP had significant stimulatory effects on transcription of the mentioned genes in comparison to that of TGFβ treated group (Fig. 1a–c). In all groups the data was obtained by normalizing the expression of the test gene to the house keeping gene (Table 1). Collagen type II was expressed in all studied groups and, qualitatively, T1 group showed higher ColII protein
Discussions
Improving the effectiveness of chondrogenic differentiation medium either by addition of cytokines or differentiation factors or by exploiting biophysico-chemical/mechanical cues has been of the focus of many researches in recent years. Given the cross talk between closely related signaling pathways, especially in tissues with overlapped differentiation pathways, it is rather likely that unwanted or side pathways occur or even overtake the desired fate, necessitating investigation of side-fates
Conclusion
It should be mentioned that although PRP significantly decreases the angiogenic and inflammatory potentials of chondrogenically differentiated cells, but it does not inhibit it completely. On the other hand, PRP enhances the mineralizatin in the matrix of the macro pellet neotissues. In addition, while the GAGs producrion increases, a sigificant portion is released. Furthermore, expression of hypertrophic marker ColX highlights that the constructed tissues under current protocols have inferior
Consent for publication
This manuscript in the present format has been approved by all authors for publication in the journal of Tissue and Cell.
Availability of data and material
Available.
Competing interest
The authors declare no conflicts of interest.
Ethics approval and consent to participate
Samples were collected from healthy patients after obtaining their written informed consent according to the ethics of Tehran University of Medical Sciences, Tehran, Iran.
Author’s contribution
H.HA and Sh. I designed the project. A.P conducted the experiments and analyzed the data. H. HA wrote the manuscript. All authors read and approved the final manuscript.
Funding
Not applicable.
Acknowledgment
This work was supported financially by Stem Cell Technology Research Center, Tehran, Iran.
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