Various forms of biohybrid robots have been developed; however, creating robots with multiple degrees of freedom remains a challenging task. In this paper, we developed a multi-joint biohybrid robot by using skeletal muscle tissue. To achieve this, we first developed a modular bio-actuator actuated by skeletal muscle tissues. The objective of this study was to enhance the contraction force of the actuator and establish optimal experimental conditions for creating high-performance robots. By applying continuous electrical stimulation for five days during culture of bio-actuator, we were able to increase the contraction force by more than threefold. Additionally, we determined the appropriate electric field based on the electrode distance, which enabled us to establish an optimal experimental setup. We also confirmed that connecting the actuators in series can significantly increase the moving distance. Connecting two actuators in series resulted in a total movement distance equivalent to the sum of the distances of each actuator. This finding suggests the potential to create robots with a larger operational workspace. Using these actuators, we first constructed a manipulator with a rotational joint. This research is expected to contribute not only to the development of various robots utilizing bio-actuators but also to advancements in biology technology.

In this paper, we fabricated a soft sensor based on PEDOT:PSS for thin film structure. The developed soft sensor can measure the contraction force at real time to be embedded in a modular bio-actuator [1]. The modular actuator generated contraction forces at 0.3 mN when applying electric pulse stimulation. To measure millinewton contraction forces and make a built in sensor, we fabricated a soft sensor using PEDOT:PSS-PDMS film. To verify that the sensor can measure the force of the actuator and can be integrated to the actuator, we analyzed characteristic of the sensor. First, we measure Young's modulus of the sensor and compare them with the bio-actuator. From the previous research [2], the Young's modulus of the bio-actuator and sensor were 45.8 kPa and 165 kPa, respectively. In addition, we simulated the sensors to estimate the change of the displacement according to the applied force. Next, we have experiments by stretching sensors using stepping motor to measure the resistance change of the sensor. From the simulation data, the displacement change is 23 µm when applying 0.3 mN of forces and then we detect the displacement change smaller than is 20 µm from the experiments. Finally, we analyzed the movement of the bio-actuator when applying stimulation using high speed camera and time response of the developed sensor. The actuator was contracted to the maximum after 150 ms from the electrical stimulation and the sensor detected the repeated motion at 10 Hz without time delay. As a result, the proposed sensor can measure the force of bioactuator at real time.

In spite of recent high attention of the biohybrid robot system, the previous researches focused on actuation system depend on simple on/off control without feedback control. To solve this problem, we proposed a soft sensor for feedback control of a bio-actuator driven by skeletal muscle. The proposed soft sensor can measure contraction forces of the proposed bio-actuator [1]. The bio-actuator was constructed with tendon structure and culture template made by polydimethylsiloxane (PDMS). It generated contraction forces at 0.3 mN when applying electrical stimulation. To measure that kind of small amount of contraction forces (0.3 mN), we fabricated a soft sensor using liquid metal, Galinstan, and HTV-2000. At first, we measured the Young’s modulus of the bioactuator and sensor and then fabricated the soft sensor having 68.52 kPa of Young’s modulus that is similar the bioactuator (45.8 kPa). Next, we simulated the sensor to estimate the resistance change according to the applied force. Since the resistance change is too small, we design the circuit to amplify the signal. Then, we detect very small resistance at milli-ohm. In addition, we analyzed time response to detect signal of actuator faster than 200 ms. As a result, the proposed sensor can measure the force of bioactuator without time delay.

In this paper, we have constructed actively perfusable multiple hepatic lobule-like vascular networks in a 3D cellular structure by using magnetic tweezers. Without well-organized channel networks, cells in a large 3D tissue cannot receive nutrients and oxygen from the channel, and therefore, the cells will be dead after few days. To construct well-organized channel networks, we fabricated a hepatic lobule like vascular networks by using magnetic fields in our previous works. However, the size of the hepatic lobule like vascular network was more than five times larger than real hepatic tissue. To improve the previous research, we have proposed several things. First, we have constructed the vascular network having similar size of the real thing in this step. Second, we have cultured the constructed structure for a long-time (more than two weeks) to verify the biocompatible condition. Third, we assemble the constructed hepatic tissues to make a large size of organ, liver. Finally, an actively perfusable system have been adopted to implement a bioreactor system by adding micro pump.

In this paper, we have fabricated a multilayered hepatic lobule-like vascular network in a 3D tissue using a heptapole magnetic tweezer. The tissue consists of cell-laden hydrogels with 3D channel networks. To fabricate multilayered channel system, magnetic hydrogel fibers were manipulated by a magnetic tweezer. The hepatic lobule tissue shows a hexagonal structure with different sizes of veins. Six portal veins transfer the blood including nutrients and oxygen to a central vein by sinusoids. The portal and central veins are made by steel rods, whereas the magnetic hydrogel fibers has a role of sinusoids. An important point of this research is to connect two veins - portal and central vein - by magnetic fibers. For this, we used magnetic tweezer with seven poles to magnetize the steel rods. In order to generate high magnetic fields, we design magnetic tweezer with a flat tip and additional lower tweezer based on simulation data. The manipulation was performed in fibrin gel inside rat liver cells. By applying high magnetic fields, we attracted magnetic fibers to the steel rods and constructed 3D channel network in cellular structure. To verify the efficiency of the channel, we supply culture medium to the channel and then analyze the cell viability according to the distance from the channel. As a result, the cells located at close to the channel show higher cell viability than others.

Fabrication of vascular network is an important research for transporting required nutrients and oxygen to the artificial tissues. In this paper, we propose a novel method to construct a hepatic lobule-like vascular network in a 3D cellular structure. The network is simply constructed by three types of veins, central vein, portal vein, and sinusoids. To realize these kinds veins, we utilize two different sizes of steel rods and magnetic fibers for delivering nutrients in 3D cellular structure. Alginate gel fibers embedding ferrite particles are prepared as the same length and are magnetized by magnetizer at 3T. A magnetic tweezer with seven poles is proposed to generate sufficient forces that can manipulate magnetized fibers. Here, two types of rods are magnetized to different magnetic poles in order to attract opposite the end of fibers. This manipulation process is performed in fibrinogen and thrombin solution with liver cells (RLC-18). After solidification of the solution, we deposit solutions with cells and fibers repeatedly, and therefore, a multi-layered structure can be constructed. In addition, we investigate cell a viability in fibrin gel according to the depth of the gel. The result is that the deeper the depth of the gel is, the lower the cell viability is. The cell viability is conducted in several condition. As a result, at the low temperature (here at 22 °C), the viability of cell is increased.

The reliable manipulation of micro-objects has been a still difficult work in scientific and technical field due to scale effects. This paper presents two types of release methods, using local stream and inertia force generated by 3D high speed motion of an end effector, for releasing and accurate positioning of biological cells. Two-fingered microhand driven by DC motors for both end effectors and PZT actuators for right end effector is employed. A parallel mechanism controlled by three PZT actuators generates 3D high speed motions to release cells adhered to one of the end effector. The local stream and inertia force created by high speed motion of the right end effector detach the cells adhered to the left end effector and right end effector, respectively. To generate the necessary external forces for separation of the attached cells, the vibration having high frequency and suitable amplitude is applied. For accurate positioning of the object, circular motions are proposed. To verify the advantage of the proposed motion, we compare five motions, three 1D motions and two circular motions. Experiments were conducted employing 16μm NIH3T3 cells. From these analyses of experiments, we conclude that the proposed motions can detach micro objects (100%) with high position accuracy (3±0.7μm) on desired position after release.

This paper presents an active release method of microobject for the improvement of the position accuracy after releasing by using 3D high speed motions of an end effector. In the micro manipulation, the release task is the challenge work due to adhesion forces. To overcome the adhesion force and to place microobject accurately on the desired location, in this paper, we propose a high speed motion by analyzing dynamic model of manipulated end effector and attached microbeads. Two fingered microhand driven by DC motors and PZT actuators is utilized for this paper. Parallel mecahnism with three PZT actuators was used for making 3D motion at high speed. To generatge high acceleration of end effector, many researchers applied simple vibration by using an additional PZT actuator. In our research, 3D high speed motion with large amplitude was achieved by only using a compacted parallel mechanism. To verify the advantage of the proposed motion, we compare five motions, 1D motions (X, Y, and Z direction) and circular motions (clockwise and counterclockwise direction), by changing the frequency and moving distance of the end effector. From these results of experiments, we conclude that the circular motion can detach microobjects with high placing accuracy after release.