Objective: To compare biocompatibility, degradation, and mechanical properties of polyglycolic acid (PGA) unwoven and woven fibers as scaffolding materials for tendon engineering in vitro. Methods: Three kinds of PGA fibers were included in this study. PGA raw material (Purac Co, Holland) was spun into single PGA filaments that were further twisted into woven fibers (PGA- 1). PGA filaments (Nantong Holycon, China) were twisted into woven fibers (PGA-2) as well. PGA-1 and PGA-2 served as experimental groups 1 and 2, while unwoven PGA fibers (Albany Co, USA) served as control group. Three types of PGA fibers were made into cord-like scaffolds that mimic tendon shape and compared with each other for biocompatibility, degradation and biomechanical properties. Avian tenocytes were isolated from digital flexor tendon and expanded in vitro. Cells of the second passage were seeded onto the PGA scaffolds. In the first 2 weeks, the cell- PGA constructs were in vitro cultured without tension and observed for cell adhesion and matrix production. The constructs were then cultured under dynamic loading in a bioreactor for another 2 weeks followed by gross and histological examinations. Results: PGA unwoven fibers have the median diameter of 10µm, while PGA-1 and PGA-2 fibers have the median diameters of 200µm and 60µm, respectively. Microscopy showed that tenocytes adhered well to all three types of PGA fibers in the first 10 days and produced abundant matrices. However, cells showed poor viability on PGA-2 fibers after 10 days, yet good viability on the other two PGA fibers over 2 weeks of observation period. H&E staining showed that there were viable cells and abundant matrices in the control and PGA-1 groups, but not in PGA-2 group after 4 weeks of in vitro culture. Additionally, PGA unwoven fibers degraded faster than woven fibers (PGA-1 and -2). Interestingly, the PGAtenocyte constructs formed tendon-like tissue after 4 weeks of in vitro culture grossly and histologically. Furthermore, mechanical test demonstrated that both PGA woven fibers had much higher tensile strength than unwoven fibers. Conclusion: Different PGA fibers have different biocompatibility with seeded tenocytes. PGA woven fibers could bear more intense mechanical loading and degrade slower than unwoven fibers, which is essential for in vitro generation of tendon tissue. Thus PGA woven fibers might serve as a proper form of scaffolding material for in vitro tendon engineering in a bioreactor.