Mouse embryonic stem cells were cultured on commercially available biodegradable macroporous microcarriers. A culture period of 1-2 weeks was needed to colonize the microcarriers. Embryonic stem cells retained their pluripotency for up to 14 days when cultured in medium supplemented with leukemia inhibitory factor. Replacing this medium by differentiation medium for 2 weeks initiated osteogenic differentiation. Encapsulation of the cell-loaded microcarriers in photopolymerizable polymers (methacrylate-endcapped poly-D,L-lactide-co-caprolactone), triacetin/hydroxyethylmethacrylate (HEMA) as solvent and with/without gelatin as porogen, resulted in a homogeneous distribution of the microcarriers in the polymer. As observed by transmission electron microscopy, viability of the cells was optimal when gelatin was omitted and when using triacetin instead of HEMA.

UMR-106 seeded microcarriers were encapsulated into in situ, photopolymerizable three-dimensional scaffolds based on d,l-lactide and epsilon-caprolactone. UMR-106 and rat bone marrow cells proliferated and differentiated well on the microcarriers. The microcarriers were completely colonized after 14 days in culture. The viscous polymer paste allowed to mix the UMR-106 seeded microcarriers and gelatin (porosigen) properly. After the photopolymerization process, microcarriers and gelatin were evenly distributed throughout the scaffold. Gelatin was leached out within 7 h, and a porous scaffold was obtained. The microcarriers remained in the scaffold even after 7 days which demonstrates that they were well entrapped in the polymer. Increasing the amount of entrapped microcarriers (20-50%) leads to scaffolds with a reduced cross-linking. Hence, the microcarriers leached out. The encapsulated UMR-106 cells did not show pyknotic nuclei which demonstrates that the photopolymerization and handling the viscous polymer/gelatin/microcarrier paste is not detrimental for the cells.

An in situ crosslinkable, biodegradable, methacrylate-endcapped poly(D,L-lactide-co-e-caprolactone) in which crosslinkage is achieved by photoinitiators was developed for bone tissue regeneration. Different combinations of the polymer with bone marrow-derived mesenchymal stem cells (BMSCs) and a-tricalcium phosphate (a-TCP) were tested in a unicortical tibial defect model in eight goats. The polymers were randomly applied in one of three defects (6.0 mm diameter) using a fourth unfilled defect as control. Biocompatibility and bone-healing characteristics were evaluated by serial radiographies, histology, histomorphometry, and immunohistochemistry. The results demonstrated cell survival and proliferation in the polymer-substituted bone defects. The addition of a-TCP was associated with less expansion and growth of the BMSCs than other polymer composites.

An in situ crosslinkable, biodegradable, methacrylate-encapped porous bone scaffold composed of D: ,L: -lactide, epsilon-caprolactone, 1,6-hexanediol and poly(ortho-esters), in which crosslinkage is achieved by photoinitiators, was developed for bone tissue regeneration. Three different polymer mixtures (pure polymer and 30% bioactive glass or alpha-tricalcium phosphate added) were tested in a uni-cortical tibial defect model in eight goats. The polymers were randomly applicated in one of four (6.0 mm diameter) defects leaving a fourth defect unfilled. Biocompatibility and bone healing properties were evaluated by serial radiographies, histology and histomorphometry. The pure polymer clearly showed excellent biocompatibility and moderate osteoconductive properties. The addition of alpha-TCP increased the latter characteristics. This product offers potentials as a carrier for bone healing promoter substances.

Sheep have many practical advantages compared with other animal models. However, their specific oral biomechanics inherent to their constant ruminant activity accounted for a high degree of the reported implant failures. Important adaptations to the implantation technique and postoperative management will be necessary to use sheep as an animal model for future oral implant related experiments.

Bone marrow cells were cultured on in situ photopolymerizable scaffolds based on D,L-lactide and epsilon-caprolactone. The influence of pore volume, size and shape were evaluated. Bone formation was demonstrated by ALP activity, osteocalcin secretion and histological analysis. TEM at the polymer interface revealed osteoblasts which secreted an extracellular matrix containing matrix vesicles loaded with apatite. Cellular infiltration was possible for scaffolds with a porosity of 70 and gelatin particle size of 250-355 microm. Scaffolds with a porosity less than 70 had the tendency to form a polymer top layer. Although increasing the gelatin particle size to 355-500 microm, leads to infiltration even in scaffolds with a porosity of 60. No infiltration was possible in scaffolds with sodium chloride as porogen. On the contrary, sucrose and gelatin leads to better interconnected scaffolds at the same porosity. Hence, spherical gelatin particles are suitable to use as porogen in photopolymerizable scaffolds.

An atomic force microscope with a specially constructed stiff cantilever was used to study the mechanical properties of ultrathin polymeric resist films devoted to use in nanoimprint lithography. The methodology, the equipment used in these studies and the results of the estimation of the Young's modulus versus temperature are presented and discussed.

The major research topic presented in this study concerns nanoindentation experiments of thin soft films, showing high adhesion forces. The aim of this research was to evaluate the nanomechanical properties of a 200 nm thick thermoplastic polymer layer mr-I 7000E devoted to nanoimprint lithography. The experiments were carried out using an atomic force microscopy instrument equipped with a modified 14 N m− 1 silicon cantilever tip (5 μm in diameter). The forces exerted during film indentation varied from 0 up to 2.5 μN. The Hertz and Johnson – Kendall – Roberts (JKR) models were used for analysis of the force – distance curves in order to extract the Young's modulus value. Contrary to the Hertz model, in the JKR model the adhesion force between tip and sample was considered as an additional load during the nanoindentation. The final results of these estimations were 0.20 and 0.25 GPa for the Hertz and JKR models, respectively. It shows that neglect of the adhesion force may cause an error of 20 %.