Downregulation of basal myosin-II is required for cell shape changes and tissue invagination.

Tissue invagination drives embryo remodeling and assembly of internal organs during animal development. While the role of actomyosin-mediated apical constriction in initiating inward folding is well established, computational models suggest relaxation of the basal surface as an additional requirement. However, the lack of genetic mutations interfering specifically with basal relaxation has made it difficult to test its requirement during invagination so far. Here we use optogenetics to quantitatively control myosin-II levels at the basal surface of invaginating cells during Drosophila gastrulation. We show that while basal myosin-II is lost progressively during ventral furrow formation, optogenetics allows the maintenance of pre-invagination levels over time. Quantitative imaging demonstrates that optogenetic activation prior to tissue bending slows down cell elongation and blocks invagination. Activation after cell elongation and tissue bending has initiated inhibits cell shortening and folding of the furrow into a tube-like structure. Collectively, these data demonstrate the requirement of myosin-II polarization and basal relaxation throughout the entire invagination process.

Osteoclast-mediated resorption primes the skeleton for successful integration during axolotl limb regeneration.

Early events during axolotl limb regeneration include an immune response and the formation of a wound epithelium. These events are linked to a clearance of damaged tissue prior to blastema formation and regeneration of the missing structures. Here, we report the resorption of calcified skeletal tissue as an active, cell-driven, and highly regulated event. This process, carried out by osteoclasts, is essential for a successful integration of the newly formed skeleton. Indeed, the extent of resorption is directly correlated with the integration efficiency, and treatment with zoledronic acid resulted in osteoclast function inhibition and failed tissue integration. Moreover, we identified the wound epithelium as a regulator of skeletal resorption, likely releasing signals involved in recruitment/differentiation of osteoclasts. Finally, we reported a correlation between resorption and blastema formation, particularly, a coordination of resorption with cartilage condensation. In sum, our results identify resorption as a major event upon amputation, playing a critical role in the overall process of skeletal regeneration.

Decreased bacterial growth on titanium nanoscale topographies created by ion beam assisted evaporation.

Titanium is one of the most widely used materials for orthopedic implants, yet it has exhibited significant complications in the short and long term, largely resulting from poor cell-material interactions. Among these many modes of failure, bacterial infection at the site of implantation has become a greater concern with the rise of antibiotic-resistant bacteria. Nanostructured surfaces have been found to prevent bacterial colonization on many surfaces, including nanotextured titanium. In many cases, specific nanoscale roughness values and resulting surface energies have been considered to be "bactericidal"; here, we explore the use of ion beam evaporation as a novel technique to create nanoscale topographical features that can reduce bacterial density. Specifically, we investigated the relationship between the roughness and titanium nanofeature shapes and sizes, in which smaller, more regularly spaced nanofeatures (specifically 40-50 nm tall peaks spaced ~0.25 µm apart) were found to have more effect than surfaces with high roughness values alone.