Plazma siromašna trombocitima kao dodatak fibroblastima kultiviranima u fibrinu obogaćenom trombocitima

Autor: Marcus Cristian Muniz Conde, Flávio Fernando Demarco, Thaís Gioda Noronha, Alissa Schmidt San Martin, Sarah Arangurem Karam, Letícia Regina Morello Sartori, Luiz Alexandre Chisini
Rok vydání: 2017
Předmět:
Zdroj: Acta Stomatologica Croatica
Acta Stomatologica Croatica, Vol 51, Iss 2, Pp 133-140 (2017)
Acta stomatologica Croatica : International journal of oral sciences and dental medicine
Volume 51
Issue 2
ISSN: 0001-7019
1846-0410
Popis: Svrha: Svrha ove studije bila je procijeniti proliferaciju i adheziju mezenhimalnih stanica (3T3/HIH) u Eagleovu mediju koji je modificirao Dulbecco (DMEM), uz dodatak plazme siromašne trombocitima (PPP) u trombocitima obogaćenom fibrinskom (PRF) nosaču. Materijal i metode: Dobivena ljudska krv obrađena je u centrifugi, uzimajući u obzir jednadžbu G = 1,12 x Rx (RPM/1000)² da bi se dobili PRF i PPP. Analize stanične adhezije i rasta provedene su MTT testom u pločici s 96 jažica uz dodatak DMEM : PPP-a (90 : 10) tijekom 24 sata. Osim toga, PRF je nanesen na pločicu s 48 jažica i 10 x 104 stanice nasađene su na svaki PRF (n = 3) s 800 ul DMEM : PPP-a (90 : 10) te kultivirane sedam dana. Obavljena je histološka analiza i imunohistokemijsko bojenje za vimentin. Rezultati: Rezultati su analizirani dvofaktorskom analizom varijance u Stata12®. Uočeno je značajno smanjenje (p < 0,05) stanične adhezije u odnosu na FBS. No prikazano je slično svojstvo rasta stanica za 10-postotni PPP (P > 0,05). Kultura fibroblasta tijekom sedam dana u PRF-u ostvarena je uz dodatak 10-postotnog PPP-a, te je pokazala pozitivno bojenje za vimentin. Dakle, stanični dodatak PPP-a smanjio je početnu adheziju stanica, ali je podržao proliferaciju adheriranih stanica i njihovu vitalnost u PRF-u. Zaključak: Ova metoda je potencijalna klinička prednost za pružanje autolognog i prirodnog nosača za staničnu kulturu u samo jednom postupku, bez uporabe ksenogenih spojeva. Mogla bi poboljšati kliničke translacijske terapije na temelju upotrebe PRF kultiviranih stanica, promovirajući regenerativni potencijal za buduću uporabu u medicini i dentalnoj medicini.
The aim of this study was to evaluate the proliferation and adhesion of mesenchymal cells (3T3/NIH) in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with Platelet-Poor Plasma (PPP) in a Platelet-Rich Fibrin (PRF) scaffold. Human blood was obtained and processed in a centrifuge considering the equation G=1.12xRx (RPM/1000)² to obtain PRF and PPP. Cell adhesion and maintenance analyses were performed by MTT assays in a 96 well plate with supplemented DMEM: PPP (90:10) for 24 hours. Besides, the PRF was deposited in a 48 well plate and 10x104 cells were seeded above each PRF (n=3) with 800μl of DMEM: PPP (90:10) and cultured for 7 days. Histological analysis and the immunohistochemical staining for Vimentin were performed. Results were analyzed by one-way ANOVA in Stata12®. A significant decrease (p0.05). Fibroblasts culture for 7 days in PRF supplemented with PPP 10% was possible, showing positive staining for Vimentin. Therefore, PPP cell supplementation decreased the initial adhesion of cells but was able to maintain the proliferation of adhered cells and able to support their viability in PRF. It seems that this method has many clinical advantages since it provides an autologous and natural scaffold with their respective supplement for cell culture by only one process, without using xenogeneic compounds. This could improve the potential of clinical translational therapies based on the use of PRF cultured cells, promoting the regenerative potential for future use in medicine and dentistry.
Databáze: OpenAIRE
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