Human osteoblasts exhibit sexual dimorphism in their response to estrogen on microstructured titanium surfaces

Background: Osseointegration is dependent on the implant surface, surrounding bone quality, and the systemic host environment, which can differ in male and female patients. Titanium (Ti) implants with microstructured surfaces exhibit greater pullout strength when compared to smooth-surfaced implants and exhibit enhanced osteogenic cellular responses in vitro. Previous studies showed that 1a,25-dihydroxyvitamin D3 [1a,25(OH)(2)D-3] has a greater effect on rat osteoblast differentiation on microstructured Ti compared to smooth Ti surfaces and tissue culture polystyrene (TCPS). The stimulatory effect of 17 beta-estradiol (E-2 ) on differentiation is observed in female osteoblasts on micro-rough Ti, but it is not known if male osteoblasts behave similarly in response to E-2 and microtopography. This study assessed whether human male and female osteoblasts exhibit sex-specific differences in response to E-2 and 1a,25(OH)(2)D-3 when cultured on microstructured Ti surfaces. Methods: Osteoblasts from three male and three female human donors were cultured on Ti discs with varying surface profiles: a smooth pretreatment (PT), a coarse grit-blasted/acid-etched (SLA), and an SLA surface having undergone modification in a nitrogen environment and stored in saline to maintain hydrophilicity (modSLA). Cells cultured on these surfaces were treated with E-2 or 1a,25(OH)(2)D-3. Results: Male and female human osteoblasts responded similarly to microstructure although there were donor-specific differences; cell number decreased, and osteocalcin (OCN), osteoprotegerin (OPG), and latent and active transforming growth factor 1 increased on SLA and modSLA compared to TCPS. Female osteoblasts had higher alkaline phosphatase activity and OCN production than male counterparts but produced less OPG. Both sexes responded similarly to 1a,25(OH)(2)D-3. E-2 treatment reduced cell number and increased osteoblast differentiation and factor production only in female cells. Conclusions: Male and female human osteoblasts respond similarly to microstructure and 1a,25(OH)(2)D-3 but exhibit sexual dimorphism in substrate-dependent responses to E-2. E-2 affected female osteoblasts, suggesting that signaling is sex-specific and surface-dependent. Donor osteoblasts varied in response, demonstrating the need to test multiple donors when examining human samples. Understanding how male and female cells respond to orthopedic biomaterials will enable greater predictability post-implantation as well as therapies that are more patient-specific.