Aida Cavallo

Giorgio Soldani

Ilenia Foffa

Marianna Buscemi

Paola Losi

Tamer Al Kayal

English

Archivio della ricerca della Scuola Superiore Sant'Anna

7 D 14 DThe bio-printed constructs using the developed bioink showed a statistically significant cellproliferation rate confirmed also by live/dead assay; fibroblasts and keratinocytes clusters diameterincreased at day 14 respect to day 7. In addition, a homogeneous viable cells distribution wasobserved in sample thickness (histological section).HOMOGENEITY TEST COMPRESSIVE MECHANICAL TESTConclusionThese results showed that the fibrinogen-based bioink (25:9 as ratiobetween sodium alginate and CaCl2 with 30 mg/ml of fibrinogen) has goodprintability properties enabling human cell proliferation and tissuematuration. The bi-layered fibrin skin construct could be used as skin graftin wound healing applications.A FIBRINOGEN-BASED BIOINK FOR SKIN TISSUE ENGINEERINGA. CavalloP,1,2, M. Buscemi2, I. Foffa2, T. Al Kayal2, G. Soldani2, P. Losi21Sant'Anna School of Advanced Study, Institute of Life Sciences, Pisa, Italy2Institute of Clinical Physiology, National Research Council, Massa, ItalyReferences1 Falanga V. Lancet, (2005). 2 Hoppenbrouwers T, et al. Thrombosis research, (2017)BackgroundThe 3D bioprinting enables to prepare skin tissue engineered grafts fornon-healing chronic wounds or skin defects leading to effect on patientsquality life1. Fibrin structural composition and binding to cells highlydetermine wound healing process2.ObjectiveThe aim is to develop a fibrinogen-based bioink with addition of alginatefor skin constructs printing followed by fibrin cross-linking using athrombin–calcium chloride (CaCl2) solution.MethodsSeveral concentrations of bovine fibrinogen (FIB-20-30-40 mg/ml) andsodium alginate were dissolved in dH2O and semi-crosslinked mixing with100mM CaCl2 solution at volumetric ratio of 25:9. The crosslinking-solutionconsists in bovine thrombin (50UT/ml) dissolved in 50mM CaCl2. Thephysical and biological properties of bioinks were characterized.The bioink that showed the best parameters was mixed with humandermal fibroblasts (6x106cells/ml) and human epidermal keratinocytes(8x106cells/ml) to print a bi-layered fibrin skin constructs. The bio-printedconstructs were cultured up to 14 days.ResultsBioink characterizationThe fibrinogen based bioinks resulted homogeneous (i.e. constant force in homogeneity test) with compressive modulus (mechanical test) thatincreases according to the fibrinogen concentration augment as well as the pores dimensions (SEM Images).012345625:9 + FIB 20 25:9 + FIB 30 25:9 + FIB 40PRE-CROSSLINKINGPOST-CROSSLINKINGSPREADING RATIODEAD0 3 D 7 DBi-layered fibrin skin construct characterizationThe 25:9+FB30 was the bioink with the lowest spreading ratio selected for biocompatibility evaluation with L929mouse fibroblast (4*106cell/ml) showing good cell proliferation (LIVE/DEAD and cell viability assays) within the 3Dprinted construct.MORPHOLOGY-SEM IMAGES25:9+FIB2025:9+FIB4025:9+FIB30AcknowledgementsThis study was supported through Dr. Vassili Fotis private donation.MASSON’STRICHROMESTAININGUnpaired t-test *P≤0,05Scale bar 100µmFibroblastKeratinocyteScale bar 100µmLIVE7 D 14 D0 7 D 14 DNHEK+NHDF CELL VIABILITY, %Unpaired t-test *P≤0,0501002003004005000 3 D 7 DL929-CELL VIABILITY, [%]0100200300400

A fibrinogen-based bioink for skin tissue engineering

https://www.iris.sssup.it/bitstream/11382/541170/1/Poster_A%20FIBRINOGEN-BASED%20BIOINK%20FOR%20SKIN%20TISSUE%20ENGINEERING.pdf

A fibrinogen-based bioink for skin tissue engineering

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