Biodegradable materials in the form of nonwoven fibers have attracted increasing attention for tissue engineering applications because they offer large surface areas and interconnected networks. In this study, cotton-like nonwoven poly(lactic acid) (PLA) fibers were successfully fabricated by centrifugal melt-spinning. The effects of centrifugal speed and secondary melt-spinning processing on the morphology, mechanical properties, and cell compatibility of the fibers were investigated. Scanning electron microscopy, differential scanning calorimetry, and Fourier-transform infrared spectroscopy (FTIR), as well as cell culturing of MC3T3-E1 were used in this study. The results showed that centrifugal speeds from 350 to 1500 rpm satisfied the needs for fiber formation. The PLA fibers we prepared had three-dimensional structures with extensive diameter distribution from the nanoscale to several tens of micrometers, large pore sizes, and high porosities, significantly different from fibers produced by electrospinning. The fiber diameters and mechanical properties could be manipulated by controlling the centrifugal speed. The finest fibers were generated at 900 rpm with average diameters of 3.47 ± 3.48 μm. The fibers created by centrifugal melt-spinning exhibited lower cytotoxicity and higher cell proliferation than those obtained by electrospinning.