Development of a micro perfusion 3-dimensional cell culture system for high-throughput biocompatibility assessment of biomaterial

• Main Authors: Cheng Yuan Lin, 林成遠
• Other Authors: M. H. Wu
• Format: Others
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/98431899005665970833

碩士 === 長庚大學 === 生化與生醫工程研究所 === 101 === Biocompatibility of ophthalmic biomaterial is known to play an important role in determining their success for application as medical devices and surgical implants. The development of in vitro biocompatibility testing model can potentially eliminate the need for animal use associated with in vivo testing model. This study aims to assess the biocompatibility of biomaterial by using a perfusion bioreactor cell culture system. Two- and Three-dimensional corneal keratocyte culture models were respectively established by cell seeding on fibronectin-coated polydimethylsiloxane or cell encapsulation within agarose gels. After a 5-day exposure to glutaraldehyde (GTA) treated gelatin discs, the cell from 3-D culture showed lower toxicity and apoptosis than those of 2-D counterparts, indicating that the cell culture model may affect biocompatibility of test materials. To further correlate the relationship between in vitro and in vivo tissue responses to the biomaterials, the GTA cross-linked gelatin discs were also implanted in an ocular anterior chamber. The results of Live/Dead and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assays showed that similar levels of death percentage and apoptotic index were observed for the groups of in vitro 3-D cultured keratocytes and in vivo stromal cells. Our findings suggest that a 3-D perfusion bioreactor cell culture system may provide an appropriate testing environment for precise determination of ocular biocompatibility of newly developed materials.