NELL-1 promotes cell adhesion and differentiation via Integrinß1.

NELL-1 (Nel-like molecule-1) is a secreted osteogenic growth factor first identified in human craniosynostosis (CS) patients. NELL-1 protein has been observed to promote bone and cartilage differentiation and to suppress adipogenesis in both in vitro and in vivo models. Despite these findings, the cell surface receptors of NELL-1 have remained unknown. In this study, we observed for the first time that NELL-1 promotes cell adherence in multiple cell lines, including ST2, C3H10T1/2, M2-10B4, ATDC5, and MC3T3 cells. Additionally, we found that NELL-1 binds to extracellular Integrinß1 and induces cell focal adhesion. By utilizing siRNA methods, we determined that NELL-1 cell surface binding and enhanced cell attachment were dependent on Integrinß1 expression. Finally, we observed that pre-coating of culture dishes or PLGA (polylactic-co-glycolic acid) scaffold with NELL-1 resulted in a significant increase in both cell attachment and osteogenic differentiation. Our results identify for the first time a cell surface target of NELL-1, Integrinß1, and elucidate new functions of NELL-1 in promoting cell adherence and osteogenic differentiation.

The incidence of orthostatic intolerance after bariatric surgery.

AIMS: Every year, over 200 000 individuals undergo bariatric surgery for the treatment of extreme obesity in the United States. Several retrospective studies describe the occurrence of orthostatic intolerance (OI) syndrome after bariatric surgery. However, the incidence of this syndrome remains unknown. MATERIALS AND METHODS: We used a prospective, de-identified registry of 4547 patients who have undergone bariatric surgery at Vanderbilt to identify cases of new-onset OI. Structured chart reviews were conducted for all subjects who reported new-onset OI post surgery. Cases of OI were confirmed using an operational case definition developed by the Vanderbilt Autonomic Dysfunction Center, and autonomic function tests results were examined for evidence of impaired autonomic function. The cumulative incidence of post-bariatric surgery OI syndrome was estimated using a life table. RESULTS: Seven hundred forty-one of 4547 (16.3%) patients included in our cohort reported new OI symptoms after surgery. After the chart review, we confirmed the presence of post-bariatric surgery OI syndrome in 85 patients, 14 with severe OI requiring pressor agents. At 5 years post surgery, follow-up is reduced to 15%; the unadjusted 5-year prevalence of OI was 1.9%. The cumulative incidence of OI syndrome adjusted for loss of follow-up was 4.2%. Most OI cases developed during weight-stable months (±5 kg). At the time of identification, 13% of OI cases showed evidence of impaired sympathetic vasoconstrictor activity. CONCLUSION: OI is frequent in the bariatric population, affecting 4.2% of patients within the first 5 years postoperatively. In 13% of post-bariatric surgery OI patients, there was evidence of impaired sympathetic vasoconstriction activity.

BMP2-induced inflammation can be suppressed by the osteoinductive growth factor NELL-1.

Bone-morphogenetic protein 2 (BMP2) is currently the only Food and Drug Administration-approved osteoinductive growth factor used in clinical settings for bone regeneration and repair. However, the use of BMP2 is encumbered by numerous clinical complications, including postoperative inflammation and life-threatening cervical swelling. Thus, methods to prevent BMP2-induced inflammation would have far-reaching clinical implications toward improving current BMP2-based methods for bone regeneration. For the first time, we investigate the potential role of the growth factor Nel-like molecule-1 (NELL-1) in inhibiting BMP2-induced inflammation. Adult rats underwent a femoral bone onlay procedure, treated with either BMP2 protein (4 mg/mL), NELL-1 protein (4 mg/mL), or both proteins combined. Animals were evaluated at 3, 7, and 14 days postoperatively by histology, histomorphometry, immunohistochemistry, and real-time PCR for markers of inflammation (TNFα, IL6). The relative levels of TNFα and IL6 in serum were also detected by ELISA. The mechanism for NELL-1's anti-inflammatory effect was further assessed through examining inflammatory markers and generation of reactive oxygen species (ROS) in the mouse embryonic fibroblast NIH3T3 cells. BMP2 significantly induced local inflammation, including an early and pronounced polymorphonuclear cell infiltration accompanied by increased expression of TNFα and IL6. Treatment with NELL-1 alone elicited no significant inflammatory response. However, NELL-1 significantly attenuated BMP2-induced inflammation by all markers and at all timepoints. These local findings were also confirmed using systemic serum inflammatory biomarkers (TNFα, IL6). In each case, NELL-1 fully reversed BMP2-induced systemic inflammation. Lastly, our findings were recapitulated in vitro, where NELL-1 suppressed BMP2 induced expression of inflammatory markers, as well as NF-κB transcriptional activity and generation of ROS. BMP2-induced inflammation is a serious public health concern with potentially life-threatening complications. In the present study, we observed that the growth factor, NELL-1, significantly attenuates or completely reverses BMP2-induced inflammation. The mechanisms of NELL-1's anti-inflammatory effect are only partially elucidated, and may include reduction of NF-κB transcriptional activity or ROS generation.