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Skottman, H. (2015). Biomimetic collagen I and IV double layer Langmuir-Schaefer films as microenvironment for human pluripotent stem cell derived retinal pigment epithelial cells. BIOMATERIALS, 51, 257-269. DOI: 10.1016/j.biomaterials.2015 AbstractThe environmental cues received by the cells from synthetic substrates in vitro are very different from those they receive in vivo. In this study, we applied the Langmuir-Schaeffer (LS) deposition, a variant of Langmuir-Blodgett technique, to fabricate a biomimetic microenvironment mimicking the structure and organization of native Bruch's membrane for the production of the functional human embryonic stem cell derived retinal pigment epithelial (hESC-RPE) cells. Surface pressure-area isotherms were measured simultaneously with Brewster angle microscopy to investigate the self-assembly of human collagens type I and IV on airsubphase interface. Furthermore, the structure of the prepared collagen LS films was characterized with scanning electron microscopy, atomic force microscopy, surface plasmon resonance measurements and immunofluorescent staining. The integrity of hESC-RPE on double layer LS films was investigated by measuring transepithelial resistance and permeability of small molecular weight substance. Maturation and functionality of hESC-RPE cells on double layer collagen LS films was further assessed by RPE-specific gene and protein expression, growth factor secretion, and phagocytic activity. Here, we demonstrated that the prepared collagen LS films have layered structure with oriented fibers corresponding to architecture of the uppermost layers of Bruch's membrane and result in increased barrier properties and functionality of hESC-RPE cells as compared to the commonly used dip-coated controls.

The photoinduced electron transfer processes were studied for hybrid systems consisting of self-assembled monolayer of zinc phthalocyanine (ZnPc) assembled on ZnO nanorods and a film of organic hole transporting material (HTM) atop. Polythiophene (P3HT) or Spiro-OMeTAD were used as HTM. The study was carried out by ultrafast transient absorption spectroscopy technique with selective excitation of ZnPc at 680 nm or P3HT at 500 nm. Data analysis revealed that photoexcitation of ZnPc in the structure ZnO|ZnPc|P3HT results in a fast (1.8 ps) electron transfer from ZnPc to ZnO, which is followed by a hole transfer from the ZnPc cation to P3HT roughly in 30 ps. However, in the case of ZnO| ZnPc|Spiro-OMeTAD structure, the primary reaction upon excitation of ZnPc is a fast (0.5 ps) hole transfer from ZnPc to Spiro-OMeTAD, and the second step is electron injection from the ZnPc anion to ZnO in roughly 120 ps. Thus, we demonstrate two structurally very similar hybrid architectures that implement two different mechanisms for photoinduced charge separation found in dye-sensitized or in organic solar cells.

Phthalocyanines (Pc) are well-known light-harvesting compounds. However, despite the tremendous efforts on phthalocyanine synthesis, the achieved energy conversion efficiencies for Pc-based dyesensitized solar cells are moderate. To cast light on the factors reducing the conversion efficiency, we have undertaken a time-resolved spectroscopy study of the primary photoinduced reactions at a semiconductor-Pc interface. ZnO nanorods were chosen as a model semiconductor substrate with enhanced specific surface area. The use of a nanostructured oxide surface allows to extend the semiconductor-dye interface with a hole transporting layer (spiro-MeOTAD) in a controlled way, making the studied system closer to a solid-state dye-sensitized solar cell. Four zinc phthalocyanines are compared in this study. The compounds are equipped with bulky peripheral groups designed to reduce the self-aggregation of the Pcs. Almost no signs of aggregation can be observed from the absorption spectra of the Pcs assembled on a ZnO surface. Nevertheless, the time-resolved spectroscopy indicates that there are inter-Pc charge separation−recombination processes in the time frame of 1−100 ps. This may reduce the electron injection efficiency into the ZnO by more than 50%, pointing out to a remaining aggregation effect. Surprisingly, the electron injection time does not correlate with the length of the linker connecting the Pc to ZnO. A correlation between the electron injection time and the "bulkiness" of the peripheral groups was observed. This correlation is further discussed with the use of computational modeling of the Pc arrangements on the ZnO surface.

Age-related macular degeneration (AMD) is the leading cause of vision loss in senior citizens in the developed world. The disease is characterised by the degeneration of a specific cell layer at the back of the eye -the retinal pigment epithelium (RPE), which is essential in retinal function. The most promising therapeutic option to restore the lost vision is considered to be RPE cell transplantation. This work focuses on the development of biodegradable biomaterials with similar properties to the native Bruch's membrane as carriers for RPE cells. In particular, the breath figure (BF) method was used to create semi-permeable microporous films, which were thereafter used as the substrate for the consecutive Langmuir-Schaefer (LS) deposition of highly organised layers of collagen type I and collagen type IV. The newly developed biomaterials were further characterised in terms of surface porosity, roughness, hydrophilicity, collagen distribution, diffusion properties and hydrolytic stability.Human embryonic stem cell-derived RPE cells (hESC-RPE) cultured on the biomaterials showed good adhesion, spreading and morphology, as well as the expression of specific protein markers. Cell function was additionally confirmed by the assessment of the phagocytic capacity of hESC-RPE.Throughout the study, microporous films consistently showed better results as cell culture materials for 2 hESC-RPE than dip-coated controls. This work demonstrates the potential of the BF-LS combined technologies to create biomimetic prosthetic Bruch's membranes for hESC-RPE transplantation.