Investigation of in vitro bone cell adhesion and proliferation on Ti using direct current stimulation

• Published in: Materials Science & Engineering C 2012-12, Vol.32 (8), p.2163-2168
• Main Authors: Bodhak, Subhadip, Bose, Susmita, Kinsel, William C., Bandyopadhyay, Amit
• Format: Article
• Language: English
• Subjects: Bone cell adhesion and proliferation; Direct current stimulation; Human osteoblast cells; Immunochemistry and confocal microscopy; MTT assay
• ISSN: 0928-4931; 1873-0191
• DOI: 10.1016/j.msec.2012.05.032

Our objective was to establish an in vitro cell culture protocol to improve bone cell attachment and proliferation on Ti substrate using direct current stimulation. For this purpose, a custom made electrical stimulator was developed and a varying range of direct currents, from 5 to 25μA, was used to study the current stimulation effect on bone cells cultured on conducting Ti samples in vitro. Cell–material interaction was studied for a maximum of 5days by culturing with human fetal osteoblast cells (hFOB). The direct current was applied in every 8h time interval and the duration of electrical stimulation was kept constant at 15min for all cases. In vitro results showed that direct current stimulation significantly favored bone cell attachment and proliferation in comparison to nonstimulated Ti surface. Immunochemistry and confocal microscopy results confirmed that the cell adhesion was most pronounced on 25μA direct current stimulated Ti surfaces as hFOB cells expressed higher vinculin protein with increasing amount of direct current. Furthermore, MTT assay results established that cells grew 30% higher in number under 25μA electrical stimulation as compared to nonstimulated Ti surface after 5days of culture period. In this work we have successfully established a simple and cost effective in vitro protocol offering easy and rapid analysis of bone cell–material interaction which can be used in promotion of bone cell attachment and growth on Ti substrate using direct current electrical stimulation in an in vitro model. ► D.C. stimulation was used to enhance in vitro bone cell adhesion and proliferation. ► Cells cultured on Ti were stimulated by using a custom made electrical stimulator. ► Optimization was performed by using a varying range of direct currents ~5 to 25μA. ► 25μA stimulation was found most beneficial for promotion of cell adhesion/growth.