Well-defined gradients in molecular alignment have been used as tools to generate large amplitude, light-induced deformations in stiff polymer networks. These systems are reversible, monolithic and based on a simple one-step self-assembly process. To fabricate the actuators, diacrylate dopants containing azobenzene moieties were blended with liquid crystalline diacrylate hosts and photopolymerized in a twisted configuration. The resulting twisted networks were heavily crosslinked with room temperature elastic moduli on the order of 1 GPa. Regardless of the temperature with respect to the glass transitions, subsequent exposure to UV radiation induced anisotropic expansion/contraction, and simple variations in geometry were used to generate uniaxial bending or helical coiling deformation modes. Because mechanical energy is directly related to elastic modulus, these systems are expected to provide significantly greater work output than contemporary polymer actuator materials.

Transplantation of multipotent mesenchymal progenitor cells is a valuable option for treating tendon disease. Tenogenic differentiation leading to cell replacement and subsequent matrix modulation may contribute to the regenerative effects of these cells, but it is unclear whether this occurs in the inflammatory environment of acute tendon disease. Equine adipose-derived stromal cells (ASC) were cultured as monolayers or on decellularized tendon scaffolds in static or dynamic conditions, the latter represented by cyclic stretching. The impact of different inflammatory conditions, as represented by supplementation with interleukin-1β and/or tumor necrosis factor-α or by co-culture with allogeneic peripheral blood leukocytes, on ASC functional properties was investigated. High cytokine concentrations increased ASC proliferation and osteogenic differentiation, but decreased chondrogenic differentiation and ASC viability in scaffold culture, as well as tendon scaffold repopulation, and strongly influenced musculoskeletal gene expression. Effects regarding the latter differed between the monolayer and scaffold cultures. Leukocytes rather decreased ASC proliferation, but had similar effects on viability and musculoskeletal gene expression. This included decreased expression of the tenogenic transcription factor scleraxis by an inflammatory environment throughout culture conditions. The data demonstrate that ASC tenogenic properties are compromised in an inflammatory environment, with relevance to their possible mechanisms of action in acute tendon disease.

Age-related degenerative changes in tendon tissue represent a common cause for acute tendon pathologies. Although the regenerative potential of multipotent mesenchymal stromal cells (MSC) was reported to restore functionality in injured tendon tissue, cellular mechanisms of action remain partly unclear. Potential tenogenic differentiation of applied MSC is affected by various intrinsic and extrinsic factors. The current study presents an in vitro model to evaluate the combined extrinsic effects of decellularized equine tendon matrix, transforming growth factor beta 3 (TGFβ3) and bone morphogenetic protein 12 (BMP12) on the tenogenic fate of equine adipose tissue-derived MSC. Monolayer MSC cultures supplemented with TGFβ3 and BMP12 as well as MSC cultured on tendon matrix scaffolds preloaded with the growth factors were incubated for 3 and 5 days. Histological evaluation and real time reverse transcription polymerase chain reaction (RT-PCR) revealed that growth factor-mediated tenogenic induction of MSC was modified by the conditions of the surrounding microenvironment. While the gene expression pattern in monolayer cultures supplemented with TGFβ3 or TGFβ3 and BMP12 revealed an upregulation for collagen 1A2, collagen 3A1, tenascin c, scleraxis and mohawk (p < 0.05), the presence of tendon matrix led to an upregulation of decorin and osteopontin as well as to a downregulation of smad8 (p < 0.05). Preloading of scaffolds with either TGFβ3, or with TGFβ3 and BMP12 promoted a tenocyte-like phenotype and improved cell alignment. Furthermore, gene expression in scaffold culture was modulated by TGFβ3 and/or BMP12, with downregulation of collagen 1A2, collagen 3A1, decorin, scleraxis, smad8 and osteopontin, whereas gene expression of tenascin c was increased. This study shows that growth factor-induced tenogenic differentiation of equine MSC is markedly altered by topographical constraints of decellularized tendon tissue in vitro. While TGFβ3 represents an effective mediator for tenogenic induction, the role of BMP12 in tenogenesis may be of modulatory character and needs further evaluation.

No abstract

The relatively small size of the conventionally evaluated SNP area is a contributory factor in high SNP parameter variability. Averaging of parameter values based on multiple CLSM frames does not necessarily result in good approximations of the respective reference values of the whole image area. This illustrates the potential for examiner bias when selecting SNP images in the central corneal area.

Membranes based on functional biocompatible polymers can be regarded as a useful model system to study biological interactions, e.g. antibody-antigen interactions or protein polymer interactions. These model systems may give a better insight into these processes and may help to find suitable polymeric structures offering biocompatibility as well as reduced polymer protein interaction. In this respect, Langmuir-Blodgett (LB) layer formation at the air/water (A/W) interface is studied in respect to polymer architecture in this article. For this purpose, narrowly distributed N-(2-hydroxypropyl)-methacrylamide (HPMA) random and block copolymers have been prepared by the RAFT polymerization method. For random copolymers different molecular weights were prepared. As for the block copolymers also the ratio of hydrophilic and hydrophobic units was varied in order to study the influence of hydrophobic block length on collapse pressure and area. The molecular weights of all polymers were around 15 kDa and 30 kDa. In the case of block copolymers we found a direct correlation of the length of the hydrophobic block to the collapse area. Furthermore, hysteresis experiments clearly point out that block copolymers form stable LB layers. No remarkable changes in collapse pressure or area could be observed. In contrast the area occupied by random copolymers changes at each hysteresis cycle indicating a loss of polymer to the aqueous subphase. In addition the LB layers were transferred onto mica substrates. The block copolymers formed stable and defect free membranes over an area of 100 microm(2) with a roughness (rms) 1.3-1.4 A. On the contrary, membranes based on random copolymers turned out to have a higher surface roughness. Our findings clearly underline the influence of polymer structure on the LB layer formation at the A/W interface.

No abstract

Transforming growth factor beta 3 (TGFβ3) promotes tenogenic differentiation and may enhance tendon regeneration in vivo. This study aimed to apply TGFβ3 absorbed in decellularized equine superficial digital flexor tendon scaffolds, and to investigate the bioactivity of scaffold-associated TGFβ3 in an in vitro model. TGFβ3 could effectively be loaded onto tendon scaffolds so that at least 88% of the applied TGFβ3 were not detected in the rinsing fluid of the TGFβ3-loaded scaffolds. Equine adipose tissue-derived multipotent mesenchymal stromal cells (MSC) were then seeded on scaffolds loaded with 300 ng TGFβ3 to assess its bioactivity. Both scaffold-associated TGFβ3 and TGFβ3 dissolved in the cell culture medium, the latter serving as control group, promoted elongation of cell shapes and scaffold contraction (p < 0.05). Furthermore, scaffold-associated and dissolved TGFβ3 affected MSC musculoskeletal gene expression in a similar manner, with an upregulation of tenascin c and downregulation of other matrix molecules, most markedly decorin (p < 0.05). These results demonstrate that the bioactivity of scaffold-associated TGFβ3 is preserved, thus TGFβ3 application via absorption in decellularized tendon scaffolds is a feasible approach.