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Endothelial cells on pet vascular prostheses impregnated with polyester-based copolymers and coated with cell-adhesive protein assemblies

Chlupac J., Filova E., Riedel T., Brynda E., Pamuła E., Lisa V., Bacakova L.

Engineering of Biomaterials

2008

Vol. 11, no. 81-84

108--111

Arterial bypass surgery with synthetic vascular prostheses achieves poor patency rates compared to autogenous natural materials, and this is a challenge for tissue engineering research concerning small caliber vascular grafts. Modifications of the prosthetic surface followed by endothelial cell seeding may reduce thrombogenicity and intimal hyperplasia. Planar polyethylene terephthalate (PET) vascular prosthetic samples were impregnated with the copolymer poly(glycolide-L-lactide) (PGL) or with the terpolymer poly(glycolide-L-lactide-(e)caprolactone) (PGLCap) in order to lower the permeability of the knitted fabrics and ensure a less adhesive background. Subsequent modification with adhesive protein assemblies composed of collagen type I (Co) in conjunction with laminin (LM), fibronectin (FN) or fibrin (Fb) gel was performed to enhance cell adhesion. Bovine pulmonary artery endothelial cells (EC) of the CPAE line were seeded on to the coatings and subjected to static tissue culture conditions for 7 days. Impregnation of the PET prostheses decreased the initial adhesion and proliferation of the EC. After coating with the protein assemblies, the impregnated PET provided better substrates for cell culture than the protein-coated PET, on which the EC population started decreasing after 4 days of culture. The cells proliferated better on the CoFN, CoFb and CoFbFN coatings than on the Co and CoLM coatings. Impregnation type and adhesive matrix protein deposition may play an important role in successful endothelialization, healing and clinical performance of vascular grafts.

Włókniste podłoża dla inżynierii tkankowej kości: hodowle komórek MG 63 w warunkach statycznych i dynamicznych

Buczyńska J., Pamuła E., Błażewicz S., Bacakova L., Parizek M., Chlupac J., Mikołajczyk T., Boguń M., Dobrzyński P.

Engineering of Biomaterials

2007

Vol. 10, no. 65-66

1-6

Resorbowalne włókna z kopolimeru L-laktydu z glikolidem (PLG) i PLG z hydroksyapatytem rozprowadzonym w ich objętości (PLG-HAP) zostały otrzymane metodą formowania z roztworu. Włókna zostały przetworzone w trójwymiarowe podłoża za pomocą metody łączenia włókien. Mikrostrukturę otrzymanych podłoży scharakteryzowano za pomocą mikroskopu stereoskopowego. Wykazano, że podłoża miały różną porowatość, wielkość i orientację pojedynczych włókien. Oddziaływanie włóknistych podłoży z komórkami kostnymi MG 63 było badane in vitro w warunkach statycznych i dynamicznych. Liczba komórek i ich morfologia były oceniane po 3 i 7 dniach od założenia hodowli. Badania wykazały że liczba komórek na materiałach włóknistych rosła wraz z czasem prowadzenia hodowli, chociaż była znacznie niższa niż na płaskiej powierzchni kontrolnej (polistyren do celów kultur komórkowych). W dynamicznych warunkach hodowli obserwowano różną proliferację komórek w zależności od rodzaju użytego podłoża: na PLG występował spadek, zaś na PLG-HAP istotny wzrost liczby komórek. Wyniki sugerują, że obecność cząstek hydroksyapatytu rozprowadzonych w objętości włókien polimerowych poprawia adhezje i proliferacje osteoblastów.

Resorbable poly(L-lactide-co-glycolide) fibres (PLG) and poly(L-lactide-co-glycolide) fibres containing hydroxyapatite nanoparticles in volume of PLG fibres (PLG-HAP) were manufactured by solution spinning process. The resultant fibres were processed into three-dimensional scaffolds using fibre bounding method. The microstructure of resorbable scaffolds was characterized by stereomicroscope. The results show that the scaffolds have different fibrous architecture including porosity, size and arrangement of individual fibres. The interaction of fibrous scaffolds with osteoblast-like MG 63 cells was tested in vitro in static and dynamic cell culture conditions. The number of adhering cells and their morphology were evaluated on days 3 and 7 after seeding. It was found that cell number increased with the cultivation time, although it was significantly lower than on control polystyrene dish (TCPS). During dynamic cultivation the number of cells decreased on PLG scaffolds, whereas on PLG-HAP scaffolds it increased. These results suggest that presence of hydroxyapatite distributed within the whole volume of resorbable polymer fibres promoted adhesion and proliferation of osteoblasts.

Human endothelium on vascular prostheses modified by extracellular matrix proteins in a flow experiment

Chlupac J., Filova E., Riedel T., Brynda E., Remy-Zolghadri M., Bareille R., Fernandez P., Daculsi R., Bordenave L., Bacakova L.

Inżynieria Biomateriałów

2006

R. 9, nr 58-60

10-13

Artificial vascular prostheses are used for bypass surgery. Thrombogenicity often cause graft occlusion. Targeted surface alterations including cell seeding may improve the haemocompatibility. Knitted commercial tubular PET (polyethylene terephtalate) vascular prostheses with collagen impregnation were modified by adsorption of laminin (LM) or coating with fibrin network (FB) on the luminal surface. Human endothelial cells were harvested, cultured and seeded at the density of 150x10\3 cells/cm square on all grafts. The cell lining was continuously visualized and quantified. The retention was 21%, 37% and only 2% of the seeding density on the unmodified (UM), LM- and FB-coated grafts, respectively. These seeded prostheses were exposed to a laminar shear stress (SS) 15 dynes/cm square for 40 minutes (UM, LM, FB) and 120 min (UM, LM) in a chamber simulating blood circulation. The SS was excluded in static (ST) control grafts. After 40 min-SS the cell numbers were78%, 27% and 72% for the UM, LM and FB prosthesis compared to the ST. The cell densities were 61% and 57% on the UM and LM after 120 min-SS. To conclude, the endothelium formed a confluent layer whereas laminin immobilization improved endothelial adhesion but not the flow resistance. Reverse effect was observed on fibrin coating.