Sealants have various applications in tissue engineering and could act…

Sealants have various applications in tissue engineering and could act as drug PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/20460822 delivery vehicles and scaffolding matrices in surgical procedures [14] and might participate in inflammatory infiltrate chemotaxis during lesion repair [15-17]. According to Buchta and colleagues?2014 Gasparotto et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Gasparotto et al. Stem Cell Research Therapy 2014, 5:78 http://stemcellres.com/content/5/3/Page 2 of[18], sealants can improve and accelerate wound-healing processes, reduce blood loss and protect against bacterial infections. Additionally, the two components of fibrin sealants can be mixed and processed in different ways [19]. Bensaid and colleagues showed that MSCs that were seeded into fibrin scaffolds and subsequently grafted into mice could create extensive networks of blood vessels and fibroblasts, a result that was not observed when the sealant alone was grafted [20]. Fibrin sealant (FS), an innovative product derived from snake venom, is produced only from animal components and thus does not transmit human blood-borne infectious diseases. Moreover, the production costs are lower than those of commercial sealants [16]. This study thus evaluated the first use of FS as a cellular scaffold and demonstrated the effects of this biomaterial on the maintenance, induction, differentiation, integrity and viability of MSCs.MethodsFibrin sealant scaffoldCommercially available fibrin sealants consist of human thrombin and fibrinogen, which might transmit infectious diseases [16]. Researchers at the Center for the Study of Venoms and Venomous Animals at UNESP (Centro de Estudos de Venenos e Animais Pe nhentos da UNESP), S Paulo State University therefore proposed a new s12998-016-0127-6 sealant produced from the thrombin-like enzyme extracted from snake venom and animal fibrinogen. The sealant was produced in this study according to the proposed standardization [15-17,21]. The product was provided in three microtubes that were stored at -20 . Upon use, the components were mixed in the previously set proportions to generate a stable clot 4-(Benzyloxy)-4-oxobutanoic acid with a dense fibrin network.Mesenchymal stem cell expansion and characterizationand CD90 (mouse anti-rat FITC; Gibco Laboratories) antibodies were used as positive markers for MSCs and a CD34 antibody (mouse anti-human FITC; BD Pharmingen, San Diego, CA, USA) was used as a negative marker. The cell suspensions were adjusted so that each test had 1 ?105 cells. The cells were distributed into five tubes (Falcon; BD Pharmingen) and were stained as follows: unstained cells (control 1), cells with only secondary antibody (control 2) and cells with either CD44, CD90 or CD34 antibodies. The antibodies of CD44, PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/10811056 CD90 and CD34 were added at a dilution of 1:10 for 30 minutes at room temperature. After antibody addition, 1 ml phosphate-buffered saline (PBS) was added to wash the cells and they were centrifuged at 2,000 rpm (620 ?g) for 10 minutes. The supernatant was discarded and the pellet resuspended in 1 ml PBS. The CD44 marker was not conjugated and a secondary antibody was then added, waiting for 30 minutes before the supernatant was discarded and the pellet resuspended in 1 ml PBS. Finally the tubes were read Ethyl 5-bromo-6-chloropicolinate on a flow cytometer (FACSCalibu.