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

Autor(es)

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

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: Molecules; v. 24, n. 15 AUG 2019.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 15/23134-8 - Influência da associação entre plasma-rico em plaquetas e ácido hialurônico na liberação de fatores de crescimento, proliferação celular e diferenciação osteogênica/condrogênica
• Beneficiário: Bruna Alice Gomes de Melo
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 16/10132-0 - Plasma-rico em plaquetas associado ao ácido hialurônico para aplicações em ortopedia regenerativa: do desenvolvimento tecnológico das preparações às caracterizações físico-químicas e biológicas in vitro
• Beneficiário: Maria Helena Andrade Santana
• Modalidade de apoio: Auxílio à Pesquisa - Regular