title: Combined effects of flow-induced shear stress and micropatterned surface morphology on neuronal differentiation of human mesenchymal stem cells.
abstract: This study investigated the combined effects of surface morphology and flow-induced shear stress on the neuronal differentiation of human mesenchymal stem cells. First, to examine the effect of surface morphology, three patterns were fabricated using photolithography and compared to the flat substrate. After selecting the most effective surface pattern, flow-induced shear stresses (0.10 and 0.25 Pa) were engaged parallel to the direction of the grooves. The degrees of alignment and neurite outgrowth were measured using digital image processing techniques for up to 10 days. Functional evaluations were also performed by monitoring the intracellular calcium concentration and the expression of synaptophysin, β-tubulin III, and MAP2. Based on these analyses, the pattern of 5 μm/5 μm/3 μm for groove/ridge/depth, respectively, was selected. Next, shear stresses (0.00, 0.10, 0.25 Pa) were applied to the cells on the selected substrate. The shear stresses affected the expression of those markers. The outgrowth measurements indicated that the shear stresses were effective at day 7. However, the effect of shear stresses tended to decrease at day 10. More cells showed higher calcium concentrations under 0.10 Pa. The alignment was also confirmed. Taken together, these results indicated that a shear stress of 0.10 Pa on the substrate of 5 μm was most effective. Therefore, such combination of mechanical stimuli and surface pattern is expected to promote neuronal differentiation with regard to functional and morphological changes.
authors.affiliation: Department of Biomedical Engineering, Inje University, Gimhae, Gyeongnam 621-749, Republic of Korea.
authors.forename: Kang Jin
authors.initials: KJ
authors.lastname: Jeon
authors.affiliation: Department of Biomedical Engineering, Inje University, Gimhae, Gyeongnam 621-749, Republic of Korea.
authors.forename: So Hee
authors.initials: SH
authors.lastname: Park
authors.affiliation: Cardiovascular and Metabolic Disease Center, Inje University, Busan 633-165, Republic of Korea.
authors.forename: Ji Won
authors.initials: JW
authors.lastname: Shin
authors.affiliation: Department of Biomedical Engineering, Inje University, Gimhae, Gyeongnam 621-749, Republic of Korea.
authors.forename: Yun Gyeong
authors.initials: YG
authors.lastname: Kang
authors.affiliation: Department of Biomedical Engineering, Inje University, Gimhae, Gyeongnam 621-749, Republic of Korea.
authors.forename: Jin-Sook
authors.initials: JS
authors.lastname: Hyun
authors.affiliation: Department of Biomedical Engineering, Inje University, Gimhae, Gyeongnam 621-749, Republic of Korea.
authors.forename: Min Jae
authors.initials: MJ
authors.lastname: Oh
authors.affiliation: Department of Biomedical Engineering, Inje University, Gimhae, Gyeongnam 621-749, Republic of Korea.
authors.forename: Seon Yeon
authors.initials: SY
authors.lastname: Kim
authors.affiliation: Department of Biomedical Engineering, Inje University, Gimhae, Gyeongnam 621-749, Republic of Korea; Cardiovascular and Metabolic Disease Center, Inje University, Busan 633-165, Republic of Korea; First Research Team/Inst. of Aged Life Redesign/UHRC, Inje University, Gimhae, Gyeongnam 621-749 Republic of Korea. Electronic address: sjw@bme.inje.ac.kr.
authors.forename: Jung-Woog
authors.initials: JW
authors.lastname: Shin
mesh_terms: D015415:Biomarkers / Q000032:analysis; D002118:Calcium / Q000032:analysis / Q000378:metabolism; D002454:Cell Differentiation; D048430:Cell Shape; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D012212:Rheology; D013314:Stress, Mechanical; D013499:Surface Properties
chemical_list: D015415:Biomarkers; D002118:Calcium
keywords: Calcium imaging; Fluid flow-induced shear stress; Functional differentiation; Mesenchymal stem cells; Micropatterned substrate; Neuronal differentiation