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01-09-2016 | Original Paper

Platelet-rich plasma (PRP) and adipose-derived mesenchymal stem cells: stimulatory effects on proliferation and migration of fibroblasts and keratinocytes in vitro

Authors: Talita Stessuk, Maria Beatriz Puzzi, Elinton Adami Chaim, Paulo César Martins Alves, Erich Vinicius de Paula, Andresa Forte, Juliana Massae Izumizawa, Carolina Caliári Oliveira, Fernando Frei, João Tadeu Ribeiro-Paes

Published in: Archives of Dermatological Research | Issue 7/2016

Abstract

The clinical use of tissue engineering associated with cell therapy is considered a new alternative therapy for the repair of chronic lesions with potential application in different medical areas, mostly in orthopedic and dermatological diseases. Platelet-rich plasma (PRP) is a rich source of growth factors and cytokines important for wound healing. Adipose-derived mesenchymal stem cells (ADSCs) have shown potential to accelerate the resolution of ulcers, to stimulate cell proliferation, and to benefit the quality of skin repair. This study aims to determine the effect of PRP and conditioned medium (CM) from ADSC on fibroblast and keratinocyte proliferation in vitro. Migration and proliferation assays were performed to evaluate the growth of fibroblasts and keratinocytes in the presence of PRP, CM, and CM + PRP. Significant proliferative stimulation was observed after 48 h of culture (p < 0.05) on mean absorbance of fibroblasts cultured with 10 and 25 % PRP, 100 % CM, and 25 % PRP + 25 % CM, if compared with control. Keratinocyte proliferation was stimulated after 48 h in cultures with 25, 50, and 100 % CM, and growth was compared with controls. The migration assay detected a significant migratory stimulus in fibroblasts cultured with 10 % PRP + 10 % CM after 48 h. These in vitro results suggest that PRP and ADSC have therapeutic potential for healing and re-epithelialization of chronic wounds in vivo.

Brem H, Sheehan P, Rosenberg HJ et al (2006) Evidence-based protocol for diabetic foot ulcers. Plast Reconstr Surg 117(7 Suppl):193S–209SCrossRefPubMed

Bunnell BA, Flaat M, Gagliard C et al (2008) Adipose-derived stem cells: isolation, expansion and differentiation. Methods 45:115–120CrossRefPubMedPubMedCentral

Chen L, Tredget EE, Wu PY et al (2008) Paracrine factors of mesenchymal stem cells recruit macrophages and endothelial lineage cells and enhance wound healing. PLoS One 3:e1886. doi:10.1371/journal.pone.0001886 CrossRefPubMedPubMedCentral

Descalzi F, Ulivi V, Cancedda R et al (2013) Platelet-rich plasma exerts antinociceptive activity by peripheral endocannabinoid-relatd mechanism. Tissue Eng Part A 19:2120–2129CrossRefPubMed

Driver VR, Hanft J, Fylling CP et al (2006) A Prospective, randomized, controlled trial of autologous platelet-rich plasma gel for the treatment of diabetic foot ulcers. Ostomy Wound Manag 52:68–87

Formigli L, Benvenuti S, Mercatelli R et al (2012) Dermal matrix scaffold engineered with adult mesenchymal stem cells and platelet rich plasma as a potential tool for tissue repair and regeneration. J Tissue Eng Regen Med 6:125–134CrossRefPubMed

Gentile P, Orlandi A, Scioli MG et al (2012) Concise review: adipose-derived stromal vascular fraction cells and platelet-rich plasma: basic and clinical implications for tissue engineering therapies in regenerative surgery. Stem Cells Transl Med 1:230–236CrossRefPubMedPubMedCentral

Hu L, Zhao J, Liu J et al (2013) Effects of adipose stem cell conditioned medium on the migration of vascular endothelial cells, fibroblasts and keratinocytes. Exp Ther Med 5:701–706PubMedPubMedCentral

Hsu CW, Yuan K, Tseng CC (2009) The negative effect of platelet-rich plasma on the growth of human cells is associated with secreted thrombospondin-1. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 107:185–192CrossRefPubMed

10.

Huang Q, Wang YD, Wu T et al (2009) Preliminary separation of the growth factors in platelet-rich plasma: effects on the proliferation of human marrow-derived mesenchymal stem cells. Clin Med J (Engl) 122:83–87

11.

Jo CH, Roh YH, Kim JE et al (2013) Optimizing platelet-rich plasma gel formation by varying time and gravitational forces during centrifugation. J Oral Implantol 39:525–532CrossRefPubMed

12.

Kakudo N, Minakata T, Mitsui T et al (2008) Proliferation-promoting effect of platelet-rich plasma on human adipose-derived stem cells and human dermal fibroblasts. Plast Reconstr Surg 122:1352–1360CrossRefPubMed

13.

Lee SH, Jin SY, Sonq JS et al (2012) Paracrine effects of adipose-derived stem cells on keratinocytes and dermal fibroblasts. Ann Dermatol 24:136–143CrossRefPubMedPubMedCentral

14.

Liang CC, Park AY, Guan JL (2007) In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2:329–333CrossRefPubMed

15.

Lucarelli E, Beccheroni A, Donati D et al (2003) Platelet-derived growth factors enhance proliferation of human stromal stem cells. Biomaterials 24:3095–3100CrossRefPubMed

16.

Marques LF, Stessuk T, Camargo IC et al (2014) Platelet-rich plasma (PRP): methodological aspects and clinical applications. Platelets 26:101–113CrossRefPubMed

17.

Mills DC, Robb IA, Roberts GC (1968) The release of nucleotides, 5-hydroxytryptamine and enzymes from human blood platelets during aggregation. J Physiol 195:715–729CrossRefPubMedPubMedCentral

18.

Park HB, Yang JH, Chung KH (2011) Characterization of the cytokine profile of platelet rich plasma (PRP) and PRP-induced cell proliferation and migration: Up regulation of matrix metalloproteinase-1 and -9 in HaCaT cells. Korean J Hematol 46:265–273CrossRefPubMedPubMedCentral

19.

Pradhan L, Nabzdyk C, Andersen ND et al (2009) Inflammation and neuropeptides: the connection in diabetic wound healing. Expert Rev Mol Med 11:e2CrossRefPubMedPubMedCentral

20.

Shai A, Maibach HI (2005) Wound healing and ulcers of the skin. Springer, Berlin

21.

Shan GQ, Zhang YN, Ma J et al (2013) Evaluation of the effects of homologous platelet gel on healing lower extremity wounds in patients with diabetes. Int J Low Extrem Wounds 12:22–29PubMed

22.

Silva Meirelles L, Caplan AI, Nardi NB (2008) In search of the in vivo identity of mesenchymal stem cells. Stem Cells 26:2287–2299CrossRefPubMed

23.

Singer AJ, Clark RA (1999) Cutaneous wound healing. N Engl J Med 341:738–746CrossRefPubMed

24.

Walter MN, Wright KT, Fuller HR et al (2010) Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays. Exp Cell Res 316:1271–1281CrossRefPubMed

25.

Wong VW, Gurtner GC (2012) Tissue engineering for the management of chronic wounds: current concepts and future perspectives. Exp Dermatol 21:729–734PubMed

26.

Xian LJ, Chowdhury SR, Saim AB et al (2015) Concentration-dependent effect of platelet-rich plasma on keratinocyte and fibroblast wound healing. Cytotherapy 17: 293–300CrossRefPubMed

27.

Xie X, Wang Y, Zhao C et al (2012) Comparative evaluation of MSCs from bone marrow and adipose tissue seeded in PRP-derived scaffold for cartilage regeneration. Biomaterials 33:7008–7018CrossRefPubMed

28.

Yang HS, Shin J, Bhang SH et al (2011) Enhanced skin wound healing by a sustained release of growth factors contained in platelet-rich plasma. Exp Mol Med 43:622–629CrossRefPubMedPubMedCentral

29.

Yoo KH, Jang IK, Lee MW et al (2009) Comparison of immunomodulatory properties of mesenchymal stem cells derived from adult human tissues. Cell Immunol 259:150–156CrossRefPubMed

30.

Zhao J, Hu L, Liu J et al (2013) The effects of cytokines in adipose stem cell-conditioned medium on the migration and proliferation of skin fibroblasts in vitro. Biomed Res Int 2013:578479PubMedPubMedCentral

Title: Platelet-rich plasma (PRP) and adipose-derived mesenchymal stem cells: stimulatory effects on proliferation and migration of fibroblasts and keratinocytes in vitro
Authors: Talita Stessuk
Maria Beatriz Puzzi
Elinton Adami Chaim
Paulo César Martins Alves
Erich Vinicius de Paula
Andresa Forte
Juliana Massae Izumizawa
Carolina Caliári Oliveira
Fernando Frei
João Tadeu Ribeiro-Paes
Publication date: 01-09-2016
Publisher: Springer Berlin Heidelberg
Published in: Archives of Dermatological Research / Issue 7/2016
Print ISSN: 0340-3696
Electronic ISSN: 1432-069X
DOI: https://doi.org/10.1007/s00403-016-1676-1

At a glance: The STEP trials

Springer Medicine

Platelet-rich plasma (PRP) and adipose-derived mesenchymal stem cells: stimulatory effects on proliferation and migration of fibroblasts and keratinocytes in vitro

Abstract

At a glance: The STEP trials

Springer Medicine

Abstract

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