This work aimed at studying the effects of dynamic culture conditions and biomimetic coating on bone cells grown on nanofiber meshes. In our previous work, biomimetic calcium phosphate coated polycaprolactone nanofibre meshes (BCP-NM) proved to be more efficient for supporting cell attachment and proliferation under static conditions, when compared to polycaprolactone nanofibre meshe (PCL-NM). However, no studies on the influence of bioreactors on the behaviour of cells cultivated on these materials were developed so far. [...]info:eu-repo/semantics/publishedVersio

Araujo, J. V.

Ashammakhi, N.

Cunha-Reis, C.

El Haj, A. J.

Gogniat, G.

Neves, N. M.

Reis, R. L.

Yang, Y.

Tissue Engineering Part A

English

Universidade do Minho: RepositoriUM

4, 7, 14, 21 and 42 days postop. qPCR was employed to determinethe expression patterns of BMPs and other molecules.Gene expressions of BMP2, BMP4, BMP7, Nog, MMP9 andMMP13 were distinctly lower in the delayed compared to thestandard healing group at several time points from day 14, whilstno differential gene expression of Coll II and Coll X was foundbetween both groups. Among the BMPs, BMP7 showed the mostmarkedly differential expression. The ﬁrst evident difference inBMP7 expression between both groups was found at day 14 sug-gesting that exogen substitution in the context of a therapeuticapproach should be postponed. The differential expression patternof bothMMP9 andMMP13 suggests that there might be a failure inendochondral ossiﬁcation in delayed bone healing.Downregulation in gene expression of osteogenic BMPs andcartilage matrix degrading MMPs may account for a considerabledelay of bone healing.(OP 87) Drug Releasing Hybrid Scaffold: New Avenue inCardiovascular Tissue EngineeringC. Spadaccio1,2, A. Rainer3, M. Chello2, E. Covino 2, M. Trom-betta3, J.A. Genovese11Cardiac Cell & Molecular Biology Lab. McGowan Institute forRegenerative Medicine and UPMC, Univ. of Pittsburgh2Department of Cardiovascular Surgery. University Campus BioMedico, Rome3CIR Bioengeneering Department. University Campus Bio Med-ico, RomeClinical application of tissue engineered vascular graft (TEVG)still needs to overcome several limitations. Our aim was to eval-uate a novel hybrid device able to simultaneously provide a spe-ciﬁc microenviroment to allow cell differentiation and to functionas a drug delivery system.We developed a multilayer hybrid construct with a middlepivotal ﬁbronectin-collagen network and two functionalized Poly-lactic acid (PLLA) layers on its inner and outer sides. The innerside layer was realized as a drug delivery system releasing Heparin(HEP) useful in the immediate management of the scaffold en-graftment, and Erythropoietin (EPO), important in differentiationand recruitment of endogenous stem cell populations.The outer onewas engrafted with TGF- for muscular differentiation.PLLA/HEP-EPO nanocomposite scaffolds were prepared byelectrospinning. Polymer solutions containing 13%w/w PLLAalone or 13%w/w PLLA with HEP and EPO solution were elec-trospun (15 kV) forming a non-woven cloth. Membranes micro-structure was evaluated by Scanning Electron Microscopy. Aftersterilization scaffolds were seeded with human mesenchymal stemcells and cell engraftment, viability, proliferation and differentia-tion were evaluated by light and confocal microscopy.Mean ﬁber diameter was 1.5 0.9 mm with a porosity favorableto cell attachment and culture. Both HEP and EPO release showedan initial burst within the ﬁrst 24 hrs and a further smoother pattern.Microscopy was consistent with a good cell attachment and via-bility with changes in cell morphology resembling to endothelialphenotype.Detection of ki67þ cells conﬁrmed cell proliferationand suggested the presence of a favourable mircorenviroment. Ashift towards CD31 positivity could be observed in the functio-nalized scaffold.(OP 88) Dynamic Mesenchymal Stem Cell Culturing for BoneTissue EngineeringS. Roeker1, S. Diederichs1, S. Boehm1, T. Scheper1, M. vanGriensven2, C. Kasper11Institut fu¨r Technische Chemie, Leibniz Universita¨t Hannover2Ludwig Boltzmann Institut fu¨r experimentelle und klinischeTraumatologie, WienMesenchymal stem cells (MSCs) are a widely used cell source intissue engineering and regenerative medicine because they can beexpanded and differentiated easily in vitro as well as in vivo. MSCscan be obtained from a variety of tissues including adipose tissue.Static cell culture with the aim of tissue engineering applications hasbeen proven to be disadvantageous since constructs grown in staticcell culture lack the mechanical stability native tissue exhibits.Mechanical stimulation has therefore become a substantial tool intissue engineering to accustom cells to their future physically activeenvironment. Moreover, differentiation of stem and progenitor cellshas been proven to be supported by mechanical strain. In this work,the effect of mechanical strain on the differentiation of a pre-osteoblastic cell line and adipose tissue derived mesenchymal stemcells was analysed. Nutrient supply especially on 3D scaffolds isheterogeneous in static cell culture leading to reduced growth ratesor even death of the tissue on the scaffold. Thus, a speciﬁcallycustomised bioreactor systems have been developed to control tissuegrowth under dynamic culture conditions. In this work cultivation ofadMSC on 3D scaffolds under controlled and reproducible condi-tions in a rotating bed system bioreactor is presented.(OP 89) Dynamic Studies of Biomimetic Coated Poly-caprolactone Nanoﬁber Meshes as Bone Extracellular MatrixAnaloguesJ.V. Araujo1,2,3, C. Cunha-Reis3, G. Gogniat3, Y. Yang3, N.Ashammakhi3, R.L. Reis1,2, A.J. El Haj3, N.M. Neves1,213B’s Research Group—Biomaterials, Biodegradables and Bio-mimetics, Department of Polymer Engineering, University ofMinho, Campus de Gualtar, Braga, Portugal2IBB—Institute for Biotechnology and Bioengineering, PT Gov-ernment Associated Laboratory, Braga, Portugal3ISTM—Institute for Science and Technology in Medicine, KeeleUniversity, Stoke-on-Trent, UKThis work aimed at studying the effects of dynamic culture con-ditions and biomimetic coating on bone cells grown on nanoﬁbermeshes. In our previous work, biomimetic calcium phosphatecoated polycaprolactone nanoﬁbre meshes (BCP-NM) proved tobe more efﬁcient for supporting cell attachment and proliferationunder static conditions, when compared to polycaprolactone na-noﬁbre meshe (PCL-NM). However, no studies on the inﬂuence ofbioreactors on the behaviour of cells cultivated on these materialswere developed so far. In fact, in vitro cultivation of constructsusing bioreactors which support efﬁcient nutrition of cells has ap-peared as an important step toward the development of functionalgrafts. In the current work, osteoblast-like cells were seeded on bothBCP-NM and PCL-NM. The formed constructs were cultured in arotating bioreactor (Synthecon, RCSS-1, USA), for different timeperiods. Cell morphology and viability were assessed by confocalORAL PRESENTATIONS 723microscopy. DNA and total protein quantiﬁcations were performedaccordingly. No signiﬁcant difference in cell proliferation wasobserved in BCP-NM and PCL-NM constructs kept under dynamicconditions when compared to the results obtained from staticstudies. However, under dynamic conditions, total protein contentswere higher on BCP-NM constructs than in the uncoated ones.Moreover, PCL-NM presented a higher number of dead cells thanBCP-NM. Obtained results point out that BCP-NM can support cellgrowth under dynamic conditions. The vast potential of using BCP-NM in applications related to bone tissue engineering was alsoconﬁrmed.Marie Curie Actions ‘‘Alea Jacta Est Training Fellowship’’ andthe European NoE EXPERTISSUES (NMP3-CT-2004-500283)supported this work.(OP 90) Dynamics and Interactions of Hemoglobin in RedBlood CellsA. Stadler1,2, K. Zerlin2, I. Digel3, G. Bueldt2, G. Zaccai1,G. Artmann31Institut Laue-Langevin, 38042 Grenoble, France2Research Centre Juelich, 52425 Juelich, Germany3University of Applied Science, 52428 Juelich, GermanyWe studied molecular dynamics and interactions of hemoglobin inwhole human red blood cells with Incoherent Elastic NeutronScattering and Small Angle Neutron Scattering. Thermal neutronwavelengths and energies match atomic distances and atomicthermal motions and allow their study in biological materialwithout damage. Protein dynamics were measured on time scalesof around 100ps which reﬂects protein side chain motions and longtime self diffusion of the whole protein.We could show that there occurs a transition of hemoglobindynamics at human body temperature of 378C in cells and solution.The measurements show that there is a break in the long term selfdiffusion coefﬁcient at human body temperature. Complementaryfrom our interaction studies, we found that hemoglobin startforming aggregated clusters at human body temperature.(OP 91) Educational Aspects of Tissue Engineering: an In-dustrial PerspectiveB.M. Thomson11Smith and Nephew Research Center, York YO10 5DF, UKTissue engineering is a branch of regenerative medicine that at-tempts to convert insights from biomedical research into noveltherapies composed of cells, growth factors and/or biocompatiblestructural materials. It aims to (i) repair, rather than replace dam-aged tissues; (ii) postpone radical surgery; and (iii) stimulate en-dogenous healing responses.Society’s need for these new clinical approaches, (e.g. biologicaltherapies for cartilage repair), is indisputable and growing in linewith the aging population. Unfortunately, the commercial andclinical success of tissue engineering has been poor to date becausethe early innovators failed to appreciate the need to resolve all thescientiﬁc, clinical, regulatory, commercial and logistical issuesinvolved in bringing a complex and unfamiliar therapy from‘bench to bedside’.Educators should therefore emphasise the multidisciplinary na-ture of regenerative medicine to students. They should stress that inorder to be successful a new productmust simultaneously resolve allthe problems associated with its introduction (e.g. (i) deﬁnition ofclinical need; (ii) cell/molecular biology of the product; (iii) struc-tural materials and biomechanics; (iv) scale-up and production; (v)patents and IPR; (vi) regulatory approval and quality control; (vii)distribution, inventory control and logistics; (viii) immunology andgraft integration; (ix) patient selection and adoption into clinicalpractice; (x) proof of cost-effectiveness and re-imbursement; and(xi) time to market and return on investors capital). Curricula,should encompass these topics and the management approaches(e.g. open innovation) required to build and maintain the multidis-ciplinary teams needed to develop regenerative medicine.(OP 92) Educational Aspects of Tissue Engineering: the Aca-demic PerspectiveD.F. Williams1,21Emeritus Professor, University of Liverpool, UK2Editor-in-Chief, BiomaterialsEveryone recognises that tissue engineering is a rapidly emerging,multidisciplinary subject that, so far, is not grounded in traditionalacademic courses. A multitude of departments, institutes andcentres concerned with tissue engineering have emerged in Uni-versities throughout the world, but almost wholly being research orclinically oriented. This has been inevitable and, as with anyemerging subject, is has been very sensible to identify its bound-aries before constructing educational courses. The time is nowcoming, however, where there is a role for new courses, in order toprovide for this education.The subject of biomaterials provides an example of how thiscould be done since 40 years ago this subject was similarly in itsinfancy. We have witnessed the introduction of courses at under-graduate and postgraduate levels, some based in medical schoolsand others in engineering schools. We cannot pretend that we havealways got this right and it is no means obvious that either theindustrial or clinical end-users of the student output have fullybeneﬁted. The most obvious question has been whether it is pos-sible or necessary to train ‘biomaterials scientists’ or for students tofollow a major relevant discipline, for example materials science,cell biology or mechanical engineering, perhaps to PhD level, andthen to develop the skills and knowledge for the transition intobiomaterials science.It is essential that the ﬁeld of tissue engineering learns theselessons; this presentation will explore the options.(OP 93) Educational Aspects of Tissue Engineering: the Per-spective of Students and Young InvestigatorsC.M. Alves1,213B’s Research Group–Biomaterials, Biodegradables and Biomi-metics, Dept. of Polymer Engineering, University of Minho,Campus of Gualtar, 4710-057 Braga, Portugal2IBB–Institute for Biotechnology and Bioengineering, PT Gov-ernment Associated Laboratory, Braga, Portugal; cmalves@dep.uminho.pt724 ORAL PRESENTATIONS

Dynamic studies of biomimetic coated polycaprolactone nanofiber meshes as bone extracellular matrix analogues

http://repositorium.sdum.uminho.pt/bitstream/1822/59210/1/TermisEU_OP89.pdf

Dynamic studies of biomimetic coated polycaprolactone nanofiber meshes as bone extracellular matrix analogues

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