Fast and Mild Strategy, Using Superhydrophobic Surfaces, to Produce Collagen/Platelet Lysate Gel Beads for Skin Regeneration
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Platelet lysate (PL) was encapsulated in collagen (Coll) millimetric gel beads, on biomimetic superhydrophobic surfaces, under mild conditions, with the aim of obtaining easy-to-handle formulations able to provide sustained release of multiple growth factors for skin ulcers treatment. The gel particles were prepared with various concentrations of PL incorporating or not stem cells, and tested as freshly prepared or after being freeze-dried or cryopreserved. Coll + PL particles were evaluated regarding degradation in collagenase-rich environment (simulating the aggressive environment of the chronic ulcers), sustained release of total protein, PDGF-BB and VEGF, cell proliferation (using particles as the only source of growth factors), scratch wound recovery and angiogenic capability. Compared to Coll solely particles, incorporation of PL notably enhanced cell proliferation (inside and outside gels) and favored scratch wound recovery and angiogenesis. Moreover, cell-laden gel particles containing PL notably improved cell proliferation and even migration of cells from one particle towards a neighbor one, which led to cell-cell contacts and the spontaneous formation of tissue layers in which the spherical gels were interconnected by the stem cells.
KeywordsCell encapsulation Collagen Growth factors Hydrogel Platelet rich plasma Scaffold
A.C. Lima acknowledges to the Portuguese Foundation for Science and Technology (FCT) for the PhD grant SFRH/BD/71395/2010. Work supported by the European Union’s Seventh Framework Programme (FP7/2007-2013) under grant agreement n° REGPOT-CT2012-316331-POLARIS, FEDER through the Competitive Factors Operation Program–COMPETE, Portugal national funds through FCT (PTDC/CTM-BIO/1814/2012), Operational Human Potential Program (POPH) developed under the scope of the National Strategic Reference Framework (QREN) from the European Social Fund (FSE), and Spain MICINN (SAF2011-22771). The authors thank Ibidi for culture inserts samples, Luis Diaz-Gomez and Patricia Días Rodríguez for the help with PL preparation and CAM assay, and Instituto de Ortopedia y Banco de Tejidos Musculoesqueléticos (Universidad de Santiago de Compostela, Spain) for the help with cell cultures.
The authors declare no potential conflicts of interest.
- 14.Hildner, F., Albrecht, C., Gabriel, C., Redl, H., & van Griensven, M. (2011). State of the art and future perspectives of articular cartilage regeneration: a focus on adipose-derived stem cells and platelet-derived products. Journal of Tissue Engineering and Regenerative Medicine, 5, e36–e51.CrossRefPubMedGoogle Scholar
- 24.Lima, A. C., Batista, P., Valente, T. A. M., Silva, A. S., Correia, I. J., & Mano, J. F. (2013). Novel methodology based on biomimetic superhydrophobic substrates to immobilize cells and proteins in hydrogel spheres for applications in bone regeneration. Tissue Engineering Part A, 19, 1175–1187.CrossRefPubMedGoogle Scholar
- 33.Smiell, J. M., Wirman, T. J., Steed, D. L., Perry, B. H., Sampson, A. R., & Schwab, B. H. (1999). Efficacy and safety of becaplermin (recombinant human platelet-derived growth factor-BB) in patients with nonhealing, lower extremity diabetic ulcers: a combined analysis of four randomized studies. Wound Repair and Regeneration, 7, 335–346.CrossRefPubMedGoogle Scholar
- 37.Kadler, K. E., Holmes, D. F., Trotter, J. A., & Chapman, J. A. (1996). Collagen fibril formation. Biochemistry Journal, 316, 1–11.Google Scholar
- 56.Yannas, I. V., Lee, E., Orgill, D. P., Skrabut, E. M., & Murphy, G. F. (1989). Synthesis and characterization of a model extracellular matrix that induces partial regeneration of adult mammalian skin. Proceedings of the National Academy of Sciences of the United States of America, 86, 933–937.CrossRefPubMedCentralPubMedGoogle Scholar