Biomimetic tissue engineering aspires to develop bioinspired implantable devices that would positively interact with the host. Given that glycosaminoglycans are involved in many physiological processes, whereas their deprivation is associated with pathophysiologies, functionalization of implantable devices with natural and/or synthetic carbohydrate moieties is at the forefront of scientific research and industrial innovation. Herein, we venture to assess the influence of various concentrations (0.01%, 0.1%, 1%) of hyaluronic acid and Ficoll on the structural, thermal, biomechanical and biological (human osteoblasts) properties of electrospun poly(lactic-co-glycolic acid) fibers. The addition of hyaluronic acid and Ficoll reduced the fiber diameter, with the 1% hyaluronic acid exhibiting the smallest fibers diameter (p < 0.001). Neither the addition of hyaluronic acid nor the addition Ficoll significantly affected the onset and peak temperatures (p > 0.05). All hyaluronic acid and Ficoll treatments significantly reduced stress at break, strain at break and elastic modulus values (p < 0.001). Hyaluronic acid and Ficoll did not affect temperatures (p > 0.05) osteoblast viability and metabolic activity; the 1% hyaluronic acid and Ficoll significantly increased (p < 0.001) osteoblast proliferation at day 21. By day 21, the 1% hyaluronic acid and 1% Ficoll fibers showed the highest alkaline phosphatase activity and calcium deposition. At day 21, osteocalcin was not detected, whereas osteopontin was detected on all samples. Collectively, our data clearly illustrate the biological benefit of nonsulfated polysaccharides as functionalization molecules.