Synthetic Heparan Sulfate Hydrogels Regulate Neurotrophic Factor Signaling and Neuronal Network Activity.

Three-dimensional (3D) synthetic heparan sulfate (HS) constructs possess promising attributes for neural tissue engineering applications. However, their sulfation-dependent ability to facilitate molecular recognition and cell signaling has not yet been investigated. We hypothesized that fully sulfated synthetic HS constructs (bearing compound 1) that are functionalized with neural adhesion peptides will enhance fibroblast growth factor-2 (FGF2) binding and complexation with FGF receptor-1 (FGFR1) to promote the proliferation and neuronal differentiation of human neural stem cells (hNSCs) when compared to constructs with unsulfated controls (bearing compound 2). We tested this hypothesis in vitro using 2D and 3D substrates consisting of different combinations of HS tetrasaccharides (compounds 3 and 4) and an engineered integrin-binding chimeric peptide (CP), which were assembled using strain-promoted alkyne-azide cycloaddition (SPAAC) chemistry. Results indicated that the adhesion of hNSCs increased significantly when cultured on 2D glass substrates functionalized with chimeric peptide. hNSCs encapsulated in 1-CP hydrogels and cultured in media containing the mitogen FGF2 exhibited significantly higher neuronal differentiation when compared to hNSCs in 2-CP hydrogels. These observations were corroborated by Western blot analysis, which indicated the enhanced binding and retention of both FGF2 and FGFR1 by 1 as well as downstream phosphorylation of extracellular signal-regulated kinases (ERK1/2) and enhanced proliferation of hNSCs. Lastly, calcium activity imaging revealed that both 1 and 2 hydrogels supported the neuronal growth and activity of pre-differentiated human prefrontal cortex neurons. Collectively, these results demonstrate that synthetic HS hydrogels can be tailored to regulate growth factor signaling and neuronal fate and activity.

Engineered glycomaterial implants orchestrate large-scale functional repair of brain tissue chronically after severe traumatic brain injury.

Severe traumatic brain injury (sTBI) survivors experience permanent functional disabilities due to significant volume loss and the brain's poor capacity to regenerate. Chondroitin sulfate glycosaminoglycans (CS-GAGs) are key regulators of growth factor signaling and neural stem cell homeostasis in the brain. However, the efficacy of engineered CS (eCS) matrices in mediating structural and functional recovery chronically after sTBI has not been investigated. We report that neurotrophic factor functionalized acellular eCS matrices implanted into the rat M1 region acutely after sTBI significantly enhanced cellular repair and gross motor function recovery when compared to controls 20 weeks after sTBI. Animals subjected to M2 region injuries followed by eCS matrix implantations demonstrated the significant recovery of "reach-to-grasp" function. This was attributed to enhanced volumetric vascularization, activity-regulated cytoskeleton (Arc) protein expression, and perilesional sensorimotor connectivity. These findings indicate that eCS matrices implanted acutely after sTBI can support complex cellular, vascular, and neuronal circuit repair chronically after sTBI.

The Anesthetic Efficacy of Articaine and Lidocaine in Equivalent Doses as Buccal and Non-Palatal Infiltration for Maxillary Molar Extraction: A Randomized, Double-Blinded, Placebo-Controlled Clinical Trial.

PURPOSE: The purpose of the present study was to evaluate the anesthetic adequacy of 4% articaine 1.8 mL versus 2% lidocaine 3.6 mL without palatal injection compared with the standard technique for the extraction of maxillary molar teeth. MATERIALS AND METHODS: This randomized, double-blinded, placebo-controlled clinical trial included patients requiring extraction of 1 maxillary molar under local anesthesia. Patients were randomly distributed into 1 of 3 groups: group A received 4% articaine 1.8 mL as a buccal injection and 0.2 mL as a palatal injection, group B received 4% articaine 1.8 mL plus normal saline 0.2 mL as a palatal injection, and group C received 2% lidocaine 3.6 mL plus normal saline 0.2 mL as a palatal injection. Pain was measured during injection, 8 minutes afterward, and during extraction using a visual analog scale. Initial palatal anesthesia and patients' satisfaction were measured using a 5-score verbal rating scale. Statistical analyses included descriptive statistics, analysis of variance, and Pearson χ2 test. Differences with a P value less than .05 were considered significant. RESULTS: Eighty-four patients were included in the study. The average pain of injection was comparable among all study groups (P = .933). Pain during extraction in the articaine group was significantly less than that experienced in the placebo groups (P < .001), although the differences between placebo groups were insignificant. Satisfaction scores were significantly higher in the articaine group compared with the placebo groups (P < .001), with comparable results between placebo groups. CONCLUSIONS: Although the anesthetic effects of single placebo-controlled buccal injections of 4% articaine and 2% lidocaine were comparable, the level of anesthetic adequacy was statistically less than that achieved by 4% articaine given by the standard technique. These results do not justify the buccal and non-palatal infiltration of articaine or lidocaine as an effective alternative to the standard technique in the extraction of maxillary molar teeth.