3D Hierarchical, Pyramid-Based Cancer Cell Chip for the Detection of Anticancer Drug Effects.

In this study, we developed a novel three-dimensional (3D) cancer cell chip using a three-floor hierarchical 3D pyramid structure (3D pyramid) to simulate 3D tumor cell growth in vitro and to detect anticancer drugs. The proposed 3D pyramidbased cancer cell chip offered substantial advantages for the agglomerate formation of tumor cells, in which cells could be maintained as tumor spheroids for up to 3 weeks. Soon after HeLa tumor cells adhered to the micropatterned pillar sidewalls, they were suspended between the pillars based on scanning electron microscopy images. Treatment with the anticancer drug oleanolic acid resulted in 46.33% and 5.86% apoptotic cells on the 2D plate and 3D pyramid-based cell chip, respectively, compared with only 0.06% apoptotic cells in the control. The increase in chemoresistance to anticancer drugs in the 3D pyramid-based cell chip might be a result of cell confluence and hypoxia due to the spheroid formation of tumor cells in the 3D pyramid structure. These results indicated that the proposed cell chip could potentially be used for anticancer drug screening or can be incorporated into other models aimed at prolonging various cell functions in culture.

Photosensitive functionalized surface-modified quantum dots for polymeric structures via two-photon-initiated polymerization technique.

In this paper, the surface modification of CdSe- and CdZnS-based quantum dots (QDs) with a functional silica shell is reported. Functionalized silica shells are prepared by two routes: either by ligand exchange and a modified Stöber process or by a miniemulsion process with amphiphilic poly(oxyethylene) nonylphenylether also know as Igepal CO-520 (IG) as oligomeric amphiphile and modified silica precursors. The polymerizable groups on the functionalized silica shell allow covalent bonding to a polymer matrix and prevent demixing during polymerization and crosslinking. This allows the homogeneous incorporation of QDs in a crosslinked polymer matrix. This paper furthermore demonstrates that the resulting QDs, which are i) shielded with a proper silica shell and ii) functionalized with crosslinkable groups, can be used in two-photon-initiated polymerization processes in combination with different photoresists to obtain highly luminescent 3D structures. The resulting luminescent structures are attractive candidates for photonics and metamaterials research.

Rotational elastic micro joint based on helix-augmented cross-spring design for large angular movement.

A new type of micro-joint based on an elastic design concept is proposed for large rotational movement. The proposed new 3D micro-joint was designed based on a cross-spring that has precise and reliable motion. However, the cross-spring has a limitation in the range of rotational angle. To improve the range of rotational movement, the proposed 3D micro-joint was modified with a helical structure. By adding the helical structure, the modified rotational joint can achieve large rotational movement The micro-joint was fabricated by the two-photon stereolithography process (TPS process). The micro-joint was manipulated by optical trapping force. With the same optical trapping force, the advantage of proposed cross-spring on the large rotational movement was evaluated. And the precise movement of the proposed micro-joint was evaluated by calculating the RMS error. It has been shown that the proposed 3D micro-joint has precise and reliable motion for large rotational angle.

Hierarchical multilayer assembly of an ordered nanofibrous scaffold via thermal fusion bonding.

A major challenge in muscle tissue engineering is mimicking the ordered nanostructure of native collagen fibrils in muscles. Electrospun nanofiber constructs have been proposed as promising candidate alternatives to natural extracellular matrix. Here, we introduce a novel method to fabricate a two-dimension (2D) sheet-type and three-dimensionally integrated nanofibrous scaffolds by combining electrospinning and rapid prototyping. The aligned 2D nanofiber mats can be processed into different configurations by the CAD/CAM-based deposition of thermally extruded microstructures. We demonstrate the feasibility of these microstructures for application in muscle tissue engineering by culturing C2C12 myoblasts and then evaluating their viability and alignment. Highly aligned cellular morphologies were successfully achieved along the direction of the nanofibers in all types of scaffolds. The hybrid scaffolds provided mechanical support and served as a topographical guide at the nanoscale, exhibiting their potential to meet the requirements for practical use in tissue engineering applications.

Synthesis and photophysical properties of two-photon absorbing spirofluorene derivatives.

New spirofluorene-based quadrupolar two-photon absorbing dyes having triphenylamine and N,N-dibutylaniline as electron donors at the end of molcules were designed and synthesized. The third-order nonlinear optical properties of these compounds were studied using a two-photon excited fluorescence method. They were found to have high two-photon absorption cross-section owing to extended conjugation of the spirofluorene moiety. The effect of varying the donor strength could be discerned by comparing the two compounds. They were successfully used as a photosensitizers for two-photon initiated polymerization of three-dimensional micro-objects.

Autofocusing method using fluorescence detection for precise two-photon nanofabrication.

We propose a method capable of focusing a laser beam on a substrate automatically via fluorescence detection from the resin of a two-photon nanofabrication system. When two-photon absorption (TPA) occurs by focusing the laser beam in the resin, fluorescence is emitted from the focusing region in the visible range. The total pixel number above the threshold value of the fluorescence images obtained by a CCD camera is plotted on a graph in accordance with the focus position. By searching for the position when the total pixel number undergoes an abrupt change in the pre-TPA region, the correct configuration of the focused laser beam can be found. Through focusing tests conducted at four vertices of a 500 µm x 500 µm square placed arbitrarily inside SCR500 resin, the errors of the autofocusing method were found to range from -100 nm to + 200 nm. Moreover, this method does not leave any polymerized marks. To verify the usefulness of the autofocusing method, the fabrication of a pyramid structure consisting of 20 layers was attempted on a coverglass. It was completely fabricated without losing a layer.

Three-dimensionally crossing manifold micro-mixer for fast mixing in a short channel length.

In this study, we report a neo-conceptive three-dimensionally (3D) crossing manifold micromixer (CMM) embedded in microchannel. Fabricated by sequential processes of photolithography and two photon absorption stereolithography, this leads to a microfluidic system with a built-in micromixer in a site controlled manner. The effectiveness of CMM is investigated numerically and experimentally. Through the numerical simulation, it is estimated that a high mixing ratio of 90% can be obtained even in a channel length shorter than five times the channel width. This compares well with the conventional passive type of micromixers that have a gradual increase in mixing efficiency with the length of the channel. Furthermore, the mixing performance of the realized CMM built-in microchannel is observed by confocal microscopy.

Proportional enlargement of movement by using an optically driven multi-link system with an elastic joint.

Diverse movements using optical manipulation have been introduced. These are generally performed in the focal region of the laser beam. To achieve a wider range of movements based on precise motion transformation, an effective method for optical manipulation that overcomes the important obstacles such as small optical trapping forces, friction, and the viscosity of fluids is required. A multi-link system with an elastic joint is introduced that provides precise motion transformation and amplification. By considering the physical properties of the structure and the optical trapping force, an elastic micron-scale joint with the simple shape of a thin plate was designed. As a further example of a multi-link system with an elastic joint, a double 4-link system for motion enlargement was designed and fabricated. By performing experimental evaluations of the fabricated structures, it was confirmed that multi-link systems with an elastic joint were effective tools for precise motion transformation through optical manipulation.

Photopatternable quantum dots forming quasi-ordered arrays.

We have functionalized core-shell CdSe/ZnS quantum dots (QDs) with a photosensitive monolayer, rendering them solution processable and photopatternable. Upon exposure to ultraviolet radiation, films composed of this material were found to polymerize, forming interconnected arrays of QDs. The photoluminescence properties of the nanocrystal films increased with photocuring. The material was found to be suitable for spin casting and was used as the active layer in a green electroluminescent device. The electroluminescence efficiency of devices containing a photocured active layer was found to be largely enhanced when compared to devices containing nonphotocured active layers. The material also showed excellent adhesion to both organic and inorganic substrates because of the unique combination of a siloxane and a photopatternable layer as ligands. The pristine functionalized nanocrystals could easily be used for two-dimensional patterning on organic and inorganic substrates. The photopatternable quantum dots were uniformly dispersed into a photopolymerizable resin to fabricate QD embedded three-dimensional microstructures.

Fabrication of three-dimensional SiC ceramic microstructures with near-zero shrinkage via dual crosslinking induced stereolithography.

Three-dimensional SiC ceramic microstructures with near-zero shrinkage were fabricated from a simple inorganic polymer mixture by inducing dual photocuring routes to produce highly dense polymer features by stereolithography and subsequent pyrolysis at 600 degrees C.

Development of dual scale scaffolds via direct polymer melt deposition and electrospinning for applications in tissue regeneration.

The objective of this study was the fabrication of highly functionalized polymeric three-dimensional (3D) structures characterized by nano and microfibers for use as an extracellular matrix-like tissue engineering scaffold. A hybrid process utilizing direct polymer melt deposition (DPMD) and an electrospinning method were employed to obtain the structure. Each microfibrous layer of the scaffold was built using the DPMD process in accordance with computer-aided design modeling data considering some structural points such as pore size, pore interconnectivity and fiber diameter. Between the layers of the three-dimensional structure, polycaprolactone/collagen nanofiber matrices were deposited via an electrospinning process. To evaluate the fabricated scaffolds, chondrocytes were seeded and cultured within the developed scaffolds for 10 days, and the levels of cell adhesion and proliferation were monitored. The results showed that the polymeric scaffolds with nanofiber matrices fabricated using the proposed hybrid process provided favorable conditions for cell adhesion and proliferation. These conditions can be attributed to enhanced cytocompatibility of the scaffold due to surficial nanotopography in the scaffold, chemical composition by use of a functional biocomposite, and an enlarged inner surface of the structure for cell attachment and growth.

Fabrication of nano- and micro-scale UV imprint stamp using diamond-like carbon coating technology.

Two-dimensional (2-D) and three-dimensional (3-D) diamond-like carbon (DLC) stamps for ultraviolet nanoimprint lithography were fabricated with two methods: namely, a DLC coating process, followed by focused ion beam lithography; and two-photon polymerization patterning, followed by nanoscale-thick DLC coating. We used focused ion beam lithography to fabricate 70 nm deep lines with a width of 100 nm, as well as 70 nm deep lines with a width of 150 nm, on 100 nm thick DLC layers coated on quartz substrates. We also used two-photon polymerization patterning and a DLC coating process to successfully fabricate 200 nm wide lines, as well as 3-D rings with a diameter of 1.35 microm and a height of 1.97 microm, and a 3-D cone with a bottom diameter of 2.88 microm and a height of 1.97 microm. The wafers were successfully printed on an UV-NIL using the DLC stamps without an anti-adhesive layer. The correlation between the dimensions of the stamp's features and the corresponding imprinted features was excellent.