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The prevalence of central nervous system (CNS) dysfunction as a result of disease or trauma remains a clinically unsolved problem which is raising increased awareness in our aging society. Human Dental Pulp Stem Cells (hDPSCs) are excellent candidates to be used in tissue engineering and regenerative therapies of the CNS due to their neural differentiation ability and lack of tumorigenicity. Accordingly, they have been successfully used in animal models of spinal cord injury, stroke and peripheral neuropathies. The ideal therapy in brain injury should combine strategies aiming to protect the damaged lesion and, at the same time, accelerate brain tissue regeneration, thus promoting fast recovery while minimizing side or long-term effects. The use of bioresorbable nanopatterned poly(lactide-co-ɛ-caprolactone) (PLCL) polymeric scaffolds as hDPCSs carriers can represent an advantage for tissue regeneration. In this chapter, we describe the surgical procedures to implant functionalized bioresorbable scaffolds loaded with hDPSCs to improve the brain lesion microenvironment in an intracranial stab wound injury model severing the rostral migratory stream (RMS) that connects the brain subventricular zone (SVZ) and the olfactory bulb in nude mice. Additionally, we also describe the technical steps after animal sacrifice for histological tissue observation and characterization., Competing Interests: Disclosures I.M.R., Y.P., B.P.R., J.L., R.B.T., D.M.A., I.R., S.M.C., J.M., L.G.S., G.I., F.L., Y.H., C.E., A.L., J.R.P. have no conflict of interest to disclose., (Copyright © 2024 Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies.)

Human Dental Pulp Stem Cells (hDPSCs) are one of the most promising stem cell sources for tissue engineering and regeneration, due to their extraordinary multi-lineage differentiation ability, ease of extraction from biological waste in dental clinics, safe non-tumorigenic phenotype, immune-tolerance upon in vivo transplantation, and great possibilities of application in autologous tissue reconstruction. The in vitro manipulation of hDPSCs paves the way for drug screening and tailor-made regeneration of damaged tissues, in the context of personalized medicine. The neural crest phenotype of these stem cells gives them the capacity to differentiate to a large variety of cell types, including neural-lineage cells. In this chapter, we describe various culture methods to generate different cellular phenotypes from hDPSCs, which can not only grow as mesenchymal-like plastic adherent cells, but also as non-adherent neurogenic dentospheres in serum-free medium. Floating dentospheres can be used to generate large populations of mature neuronal and glial marker expressing cells, which may be cultured over a substrate of nanopatterned scaffold based on biodegradable poly(lactide-co-caprolactone) (PLCL) to induce a controlled alignment of neurites and cell migration, to generate in vivo biocompatible constructs for nerve tissue bioengineering., Competing Interests: Disclosures A.L., J.R.P., J.L., Y.P., B.P.R., P.G.G., V.U.E. and G.I. have no conflicts of interest to disclose. J.R.S. declares consulting/advisory fees from Polimerbio S.L., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Quitting smoking is key for patients with Chronic Obstructive Pulmonary Disease (COPD). Standard recommendations for quitting smoking are implemented for COPD as well. Varenicline Tartrate (VT) is the most effective drug to help quit smoking, but few studies have analysed its effectiveness., Aim of the Study: To determine the Abstinence Rate (AR) at 12 months, in COPD and non-COPD smokers., Methods: Observational study in 31 COPD (post bronchodilator-BD FEV 1 /FVC <0.70) and in 63 non-COPD smokers, were invited to receive treatment with Varenicline Tartrate (VT). Fourteen subjects with COPD and 46 non-COPD subjects received additionally Cognitive-Behavioral Therapy (CBT). Abstinence rate (AR) was validated by exhaled carbon monoxide CO (COe), in addition to a phone or face-to-face interview. Motivation score was measured with a visual analogue scale (MS)., Results: Differences between COPD and non-COPD, mean FEV 1 /FVC ratio 0.52 ± 0.10 vs. 0.90 ± 0.15, age 60 ± 10 vs. 47 ± 10 years, smoking pack-years 37 ± 3.5 vs. 22 ± 12, and COe 16 ± 11 vs. 12 ± 9 ppm were statistically significant (p < 0.05); for MS the score was 93 ± 11 vs. 93 ± 11 and for attempts to quit (AQ) 2 ± 2 vs. 2 ± 3 were not. AR was not significantly different at 12 months (61.2 vs. 42.8% p = 0.072). Motivation was the only significant one-year AR predictor., Conclusions: COPD smokers had a similar response (higher tendency) to VT regardless of the presence of airflow obstruction and stronger nicotine addiction., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The implementation of cell replacement therapies for Huntington's disease using multipotent neural stem cells (NSCs) requires the specific differentiation into gamma-aminobutyric acid (GABA) neuronal subtype before transplantation. Here we present an efficient culture procedure that induces stable GABAergic neurons from the immortalized striatal neural stem cell line ST14A. This process requires sequential retinoic acid treatment and KCl depolarization. Initial addition of 10 microM retinoic acid increased cell survival and promoted neuronal differentiation. Subsequent stimulation with 40 mM KCl induced specific differentiation into GABAergic neurons, yielding 74% of total cultured cells. KCl-evoked Ca(2+) influx reduced cell proliferation and nestin expression, and induced neurite outgrowth and GABAergic markers as well as GABA contents, release, and uptake. Characterization of the integration, survival, and phenotype of these predifferentiated GABAergic neurons following transplantation into the adult brain in a model of Huntington's disease revealed long-term survival in quinolinate-lesioned striata. Under these conditions, cells maintained their GABAergic phenotype and elaborated neurite processes with synaptic contacts with endogenous neurons. In conclusion, we have generated a homogeneous population of functional GABAergic neurons from a neural stem cell line, which survive and maintain their acquired fate in vivo. These data may lend support to the possibility of cell replacement therapies for Huntington's disease using neural stem cells.