Physical Profile and Impact of a Calcium-Incorporated Implant Surface on Preosteoblastic Cell Morphologic and Differentiation Parameters: A Comparative Analysis.

PURPOSE: To assess and compare topographic features and preosteoblastic cell responses of a new hydrothermally treated, calcium-incorporated surface against other commercially available implant surfaces. MATERIALS AND METHODS: Four different surfaces were the subject of comparison in this study: machined (MC), resorbable blast media (RBM), sandblasted/large-grit/acid-etched (SLA), and calcium-incorporated SLA (Ca-SLA). Surface morphology and roughness were first characterized by scanning electron microscope (SEM) and white light interferometer, respectively. Preosteoblastic MC3T3-E1 cells were then cultured on the titanium surfaces. Cell morphology was observed at 24 hours, 48 hours, 7 days, and 15 days by SEM; differentiation was assessed at 7, 11, and 15 days by assaying alkaline phosphatase (ALP) activity and osteocalcin (OCN) levels. RESULTS: Surface characterization revealed nanotopographic features on Ca-SLA. At topographic analysis, SLA and Ca-SLA showed similar roughness values. Significant differences in cell differentiation parameters were found only at 15 days between the SLA surfaces (both Ca-incorporated and nonincorporated) and MC. CONCLUSION: Collectively, this study demonstrated that hydrothermal treatment determines the formation of nanotopography without altering the SLA microtopography. Moreover, Ca-SLA and SLA induce MC3T3-E1 cell differentiation at comparable levels.

Anti-CD20 Therapy Acts via FcγRIIIA to Diminish Responsiveness of Human Natural Killer Cells.

Natural killer (NK) immune cells mediate antibody-dependent cellular cytotoxicity (ADCC) by aggregating FcγRIIIA/CD16, contributing significantly to the therapeutic effect of CD20 monoclonal antibodies (mAb). In this study, we show that CD16 ligation on primary human NK cells by the anti-CD20 mAb rituximab or ofatumumab stably impairs the spontaneous cytotoxic response attributable to cross-tolerance of several unrelated NK-activating receptors (including NKG2D, DNAM-1, NKp46, and 2B4). Similar effects were obtained from NK cells isolated from patients with chronic lymphocytic leukemia in an autologous setting. NK cells rendered hyporesponsive in this manner were deficient in the ability of these cross-tolerized receptors to phosphorylate effector signaling molecules critical for NK cytotoxicity, including SLP-76, PLCγ2, and Vav1. These effects were associated with long-lasting recruitment of the tyrosine phosphatase SHP-1 to the CD16 receptor complex. Notably, pharmacologic inhibition of SHP-1 with sodium stibogluconate counteracted CD20 mAb-induced NK hyporesponsiveness, unveiling an unrecognized role for CD16 as a bifunctional receptor capable of engendering long-lasting NK cell inhibitory signals. Our work defines a novel mechanism of immune exhaustion induced by CD20 mAb in human NK cells, with potentially negative implications in CD20 mAb-treated patients where NK cells are partly responsible for clinical efficacy.