Surface roughness modulates the response of MG63 osteoblast-like cells to 1,25-(OH)2D3 through regulation of phospholipase A2 activity and activation of protein kinase A

C. H. Lohmann, R. Sagun, V. L. Sylvia, D. L. Cochran, D. D. Dean, B. D. Boyan, Z. Schwartz

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79 Scopus citations


Implant surface roughness influences osteoblast proliferation, differentiation, and local factor production. Moreover, the responsiveness of osteoblasts to systemic hormones such as 1,25-(OH)2D3 is altered by the effects of surface roughness; on the roughest Ti surfaces the effects of roughness and 1,25-(OH)2D3 are synergistic. Prostaglandin E2 (PGE2) appears to be involved in mediating the effects of surface roughness on the cells, as well as in the response to 1,25-(OH)2D3. However, it is not yet known through which signaling pathways surface roughness exerts its effects on the response of osteoblasts to 1,25-(OH)2D3. The present study examined the potential role of protein kinase A (PKA), phospholipase A2(PLA2), and protein kinase C (PKC) in this process. MG63 osteoblast-like human osteosarcoma cells were cultured on cpTi disks with R(a) values of 0.54 μm (PT), 4.14 μm (SLA), or 4.92 μm (TPS). PKA was inhibited by adding H8 to the cultures; similarly, PLA2 was inhibited with quinacrine or activated with melittin, and PKC was inhibited with chelerythrine. Inhibitors or activators were included in the culture media through the entire culture period or for the last 24 h of culture. In addition, cultures were treated for 24 h with inhibitors or activators in the presence of 1,25-(OH)2D3. The effects on cell number and alkaline phosphatase specific activity were determined after 24 h; PKC activity was determined after 9 min and at 24 h. Cell number was reduced on rough surfaces, and alkaline phosphatase activity was increased. 1,25-(OH)2D3 had a synergistic effect with surface roughness on alkaline phosphatase. However, neither surface roughness nor 1,25- (OH)2D3 had an effect on PKC. H8 treatment for 24 h inhibited cell number and alkaline phosphatase on all surfaces; however, when it was present throughout the culture period, the PKA inhibitor had no effect on cell number, but decreased alkaline phosphatase-specific activity. H8 reduced the 1,25(OH)2D3-mediated effect on cell number and alkaline phosphatase. Quinacrine inhibited cell proliferation and alkaline phosphatase on all surfaces and further reduced the 1,25(OH)2D3-dependent decreases in both parameters. Melittin had no effect when applied for 24 h and did not modify the 1,25-(OH)2D3 effect; however, when present throughout the culture period, it caused a decrease in proliferation and an increase in enzyme activity. Chelerythrine, the PKC inhibitor, only inhibited cell proliferation when it was present throughout the entire culture period. However, it decreased alkaline phosphatase in cultures treated for 24 h, but increased enzyme activity when it was present for the entire culture period. The results indicate that surface roughness and 1,25-(OH)2D3 both mediate their effects through PLA2 which catalyzes the rate-limiting step in PGE2 production. Further downstream, PGE2 activates PKA. Surface roughness- dependent effects are also mediated through PKC, but only after the cells have reached confluence and are undergoing phenotypic maturation. The effect of surface roughness on responsiveness to 1,25-(OH)2D3 is mediated through PLA2/PKA and not through PKC.

Original languageEnglish (US)
Pages (from-to)139-151
Number of pages13
JournalJournal of Biomedical Materials Research
Issue number2
StatePublished - 1999


  • 1,25(OH)D
  • In vitro
  • Osteoblasts
  • Phospholipase A
  • Protein kinase A
  • Protein kinase C
  • Surface roughness
  • Titanium

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials


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