Centrifugation Conditions in the L-PRP Preparation Affect Soluble Factors Release and Mesenchymal Stem Cell Proliferation in Fibrin Nanofibers

Author(s)

Gomes de Melo, Bruna Alice ^[1] ; Malheiros Luzo, Angela Cristina ^[2] ; Santos Duarte Lana, Jose Fabio ^[3] ; Andrade Santana, Maria Helena ^[1]

Affiliation

^[1] Univ Estadual Campinas, Sch Chem Engn, Dept Engn Mat & Bioproc, BR-13083852 Campinas, SP - Brazil

^[2] Univ Estadual Campinas, Umbil Cord Blood Bank, Hematol & Hemotherapy Ctr, BR-13083878 Campinas, SP - Brazil

^[3] Bone & Cartilage Inst, BR-13334170 Indaiatuba, SP - Brazil

• Document type: Journal article
• Source: Molecules; v. 24, n. 15 AUG 2019.
• Web of Science Citations: 0

Abstract

Leukocyte and platelet-rich plasma (L-PRP) is an autologous product that when activated forms fibrin nanofibers, which are useful in regenerative medicine. As an important part of the preparation of L-PRP, the centrifugation parameters may affect the release of soluble factors that modulate the behavior of the cells in the nanofibers. In this study, we evaluated the influences of four different centrifugation conditions on the concentration of platelets and leukocytes in L-PRP and on the anabolic/catabolic balance of the nanofiber microenvironment. Human adipose-derived mesenchymal stem cells (h-AdMSCs) were seeded in the nanofibers, and their viability and growth were evaluated. L-PRPs prepared at 100x g and 100 + 400x g released higher levels of transforming growth factor (TGF)-beta 1 and platelet-derived growth factor (PDGF)-BB due to the increased platelet concentration, while inflammatory cytokines interleukin (IL)-8 and tumor necrosis factor (TNF)-alpha were more significantly released from L-PRPs prepared via two centrifugation steps (100 + 400x g and 800 + 400x g) due to the increased concentration of leukocytes. Our results showed that with the exception of nanofibers formed from L-PRP prepared at 800 + 400x g, all other microenvironments were favorable for h-AdMSC proliferation. Here, we present a reproducible protocol for the standardization of L-PRP and fibrin nanofibers useful in clinical practices with known platelet/leukocyte ratios and in vitro evaluations that may predict in vivo results. (AU)

• FAPESP's process: 15/23134-8 - Influence of the association between platelet-rich plasma and hyaluronic acid on the release of growth factors, cellular proliferation and osteogenic/ chondrogenic differentiation
• Grantee: Bruna Alice Gomes de Melo
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/10132-0 - Platelet-rich plasma associated to hyaluronic acid for applications in regenerative orthopedics: from the technological development of preparations to physicochemical and in vitro biological characterizations
• Grantee: Maria Helena Andrade Santana
• Support Opportunities: Regular Research Grants