Stem cell-based tissue engineering in treating intervertebral disc (IVD) degeneration is promising. An appropriate cell scaffold can maintain the viability and function of transplanted cells. Injectable hydrogel has the potential to be an appropriate cell scaffold as it can mimic the condition of the natural extracellular matrix (ECM) of nucleus pulposus (NP) and provide binding sites for cells. This study was aimed at investigating the effect of injectable hydrogel-loaded NP-derived mesenchymal stem cells (NPMSC) for the treatment of IVD degeneration (IDD) in rats. In this study, we selected injectable 3D-RGD peptide-modified polysaccharide hydrogel as a cell transplantation scaffold. In vitro, the biocompatibility, microstructure, and induced differentiation effect on NPMSC of the hydrogel were studied. In vivo, the regenerative effect of hydrogel-loaded NPMSC on degenerated NP in a rat model was evaluated. The results showed that NPMSC was biocompatible and able to induce differentiation in hydrogel in vivo. The disc height index (almost 87%) and MRI index (3313.83±227.79) of the hydrogel-loaded NPMSC group were significantly higher than those of other groups at 8 weeks after injection. Histological staining and immunofluorescence showed that the hydrogel-loaded NPMSC also partly restored the structure and ECM content of degenerated NP after 8 weeks. Moreover, the hydrogel could support long-term NPMSC survival and decrease cell apoptosis rate of the rat IVD. In conclusion, injectable hydrogel-loaded NPMSC transplantation can delay the level of IDD and promote the regeneration of the degenerative IVD in the rat model.

Background: Numerous complications of lumbar fusion surgery have been reported, with adjacent segment disease (ASD) being one of the most important. Few studies describe the effect of sagittal, horizontal screw angles and distance of pedicle screw apex to superior endplate on the incidence of ASD in lumbar spine. The purpose of this retrospective study is to evaluate the hypothesis that unsatisfactory pedicle screw insertion positions would increase the likelihood of ASD. Methods: Outpatients with lumbar spinal stenosis underwent posterolateral lumbar fusion at L4-S1 with a least 2-year follow-up were studied. ASD at L3–L4 was defined as a condition in which intervertebral disk narrowing, posterior vertebral opening, and vertebral slippage progress at the last follow-up in comparison with the postoperative. Independent t test was performed to compare data between two groups; Spearman analysis was performed to analyze the relationship between two continuous variables. Multivariate binary logistic models were performed to identify the independent risk factors of ASD. The receiver operating characteristic (ROC) curve was performed to measure model discrimination and Hosmer–Lemeshow (H–L) test was used to measure calibration. ROC curve evaluated the discrimination ability of sagittal screw angle and distance in predicting incidence of ASD. Results: Patients in ASD group exhibit higher incidence of osteoporosis, higher Visual analogue scale (VAS), Oswestry disability index (ODI), bigger sagittal screw angle, shorter distance of pedicle screw apex to superior endplate than those in non-ASD group (p < 0.05). VAS, ODI at the last follow-up were positively correlated with Pfirrmann grade of L3–4 disk and sagittal screw angle, while negatively correlated with distance of screw apex to superior endplate (p < 0.05). Multivariate binary logistic model indicated that follow-up time (odds ratio [OR] 1.637, 95% confidence interval [CI] 1.186–2.260), distance of screw apex to superior endplate (OR 0.150, 95% CI 0.067–0.336), sagittal screw angle (OR 2.404, 95% CI 1.608–3.594) were statistically significant. The models showed great discrimination and calibration. The area under the curve of ASD identified by sagittal angle and distance was 0.895 and the cut-off values were 5.500° and 6.250 mm, respectively. Conclusion: Sagittal screw angle and distance of screw apex to superior endplate were significantly associated with the risk of ASD.

Background: Apart from the current understanding of enzyme function, the mechanism of ectonucleotide pyrophosphatase/phosphodiesterase 1 (Enpp1) deficiency-associated osteoporosis is unknown. We aimed to explore the changes in the expression of signaling pathways of bone tissues involved in Enpp1 deficiency. Methods: The body weights and morphology and histology of the bones of male Enpp1 knockout (KO) and wild-type (WT) mice were assessed. The humeri of WT and Enpp1 KO mice at 12 weeks of age were subjected to high-throughput quantitative molecular measurements, and bioinformatics analysis was performed. Proteins from humeri and calvarial pre-osteoblasts (Pobs) were used to verify the differentially expressed signaling pathways and to explain the mechanism of Enpp1 deficiency-associated osteoporosis. Results: Enpp1 KO mice had significantly lower body weight and trabecular bone mass in the hindlimbs than WT mice. Proteomics and immunoblotting showed that Enpp1 deletion downregulated the expression of the p38 mitogen-activated protein kinase (MAPK) signaling pathway in bones. Lysophosphatidic acid (LPA) was involved in activating the MKK3/p38 MAPK/PCNA pathway and proliferating Pobs in Enpp1 KO mice, whereas a p38 MAPK inhibitor suppressed the LPA-induced pro-proliferation phenotype (p < 0.05). Conclusion: The inhibition of MKK3/p38 MAPK/PCNA pathway plays an important role in the development of osteoporosis caused by Enpp1 deficiency, and LPA partially rescued the proliferation of pre-osteoblasts via the MKK3/p38 MAPK/PCNA pathway.

An ultrasound-responsive silk fibroin (SF)-graphene oxide (GO)-based hydrogel (SF/GO-gel) was created in this study to facilitate the sustained delivery of troxerutin (Trox) in order to alleviate intervertebral disc degeneration (IDD). The SF/GO-gel@Trox system exhibited exceptional biocompatibility, mechanical robustness, and controlled drug release. In vivo, X-ray and MRI demonstrated that SF/GO-gel@Trox substantially preserved disc height index and hydration in comparison to IDD. Histology also confirmed the preservation of extracellular matrix expression. In a mechanistic manner, SF/GO-gel@Trox activated tyrosine-protein kinase receptor (Tie2), which subsequently promoted Casitas B-lineage Lymphoma (CBL)-mediated K48-linked ubiquitination and degradation of epidermal growth factor receptor (EGFR). Consequently, NF-κB-driven inflammation and senescence were suppressed. The protective effects of SF/GO-gel@Trox were demonstrated by transcriptomics and functional assays to be underpinned by Tie2/PI3K-Akt signaling, whereas degeneration was exacerbated by Tie2 knockdown. It is important to note that SF/GO-gel@Trox stabilized phosphorylated Tie2 (Y992), which in turn improved the interaction between CBL and EGFR, thereby accelerating the turnover of EGFR. In a rat IDD model, Tie2 overexpression was able to mitigate disc structural damage through hydrogel delivery, whereas concurrent EGFR expression reversed these benefits. The SF/GO-gel platform facilitated the localized, ultrasound-triggered release of Trox, providing a novel approach to IDD therapy that targets the Tie2/EGFR axis. These results emphasize the potential of SF/GO-gel@Trox as a multifunctional system that can effectively combat IDD by utilizing coordinated anti-inflammatory, pro-anabolic, and anti-catabolic mechanisms.