Using rapid prototyping technology, three-dimensional (3D) structures composed of hepatocytes and gelatin hydrogel have been formed. This technique employs a highly accurate 3D micropositioning system with a pressure-controlled syringe to deposit cell/biomaterial structures with a lateral resolution of 10 µm. The pressure-activated micro-syringe is equipped with a fine-bore exit needle for which a wide variety of 3D patterns with different arrays of channels (through-holes) were created. More than 30 layers of a hepatocyte/gelatin mixture were laminated into a high spacial structure using this method. The laminated hepatocytes remained viable and performed biological functions in the construct for more than 2 months. The rapid prototyping technology offers potential for eventual high-throughout production of artificial human tissues or organs.

There is a need for rapid fabrication of tissue or organs with well-defined structures and functions in regenerative medicine. Two patterns of cell/matrix constructs containing hepatic cells, gelatin and fibrinogen were successfully created by automated rapid prototyping techniques and stabilized with thrombin. No apparent cell damage was found during the process. Mechanical characterization demonstrated that a 1:1 ratio gelatin/fibrin mixture had the greatest elasticity modulus and compressive strength. Microscopic and histological observations showed that hepatic cells were embedded in the gelatin/fibrinogen matrix and were proliferating. Immunostaining and biochemical analysis indicated that the embedded hepatocytes secreted albumin. Fibrin appears to be a favorable component for a gelatin based cell assembly matrix in that it is bioresorbable, easily manipulated, and supports in vitro cell functions.

Human adipose-derived stem cells (hADSC) encapsulated in alginate and alginate/gelatin microspheres with adjustable properties were fabricated via an improved microsphere generating device. The mechanism of the device, porous property, swelling behavior of the microspheres and hADSC proliferation as well as adipogenic differentiation were studied extensively. Microspheres with high-ratio evenly distributed adipocytes could be obtained by utilizing the proper matrix material and manufacturing parameters. The adipocyte/hADSC microspheres were a sound in vitro mimicking of a natural fat lobule and therefore a good candidate for adipose tissue engineering and regenerative medicine.

An organ manufacturing technique was developed that enables the formation of cell/extracellular matrix (ECM) complexes for in vitro or in vivo growth. In this study, a three-dimensional (3D) structure composed of hepatocytes and gelatin/alginate hydrogel was made using a cell assembler-I apparatus to thoroughly control cell assembling. Hepatocytes and ECM were constructed into 10 X 10 X 3mm³ structures according to a designed pattern. The embedded hepatocytes remained viable and performed biological functions in the construct for more than 12 days. This 3D structure has the potential to be used as a precursor for tissue or organ regeneration. This technology offers the potential for high-throughput production of artificial human tissues and organs.

Brain tissue engineering has now emerged as one of the most promising treatments for the traumatic brain injury. In this article, two groups of three-dimensional (3D) hydrogel structures composed of gelatin and gelatin/hyaluronan have been formed using our 3D cell assembly technique for in vivo study in rats, in order to investigate their effects in reparation of injury in the central nervous system (CNS). The structures were implanted into cortical defects created in rat brains, and their abilities to improve the brain tissue reconstruction were then evaluated. After 4, 8, 10, and 13 weeks of implantation, sections of brains were processed with NISSL staining for observing the immigration of host neural cells into the implanted materials and the degradation property of the materials. The results showed that simplex gelatin and gelatin/hyaluronan (20:1) with 3D structures both have good biocompatibility with brain tissue while gelatin/hyaluronan has a better contiguity with the surrounding tissue. Through our primary study, it seems that 3D gelatin/hyaluronan structures may be useful in brain tissue repair.