Background/Objectives: Ischemic stroke is a leading cause of death and disability, and neuroprotection therapies, or those that increase recovery, are not available. While the garlic-derived bioactive compound S-allyl cysteine (SAC) has shown neuroprotective properties, its subacute long-term effects remain underexplored, particularly in females. Methods: We evaluated whether SAC supports functional recovery after ischemia/reperfusion (IR), focusing on neurotrophin signaling, tropomyosin receptor kinase B (TrkB), protein kinase B (AKT), and extracellular signal-regulated kinase (ERK). Adult female Wistar rats underwent 1 h of ischemia and 15 days of reperfusion. SAC (100 mg/kg, i.p.) was administered at the onset of reperfusion and daily for 15 days. Motor and cognitive deficit tests were performed. Infarct area, Ki67, brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), nerve growth factor (NGF), pTrkB, pAKT, and pERK levels were quantified in the cortex, striatum, and hippocampus. Results: MicroPET analysis revealed comparable glucose uptake between the IR and IR + SAC groups, indicating similar ischemic severity. SAC reduced infarct area (54.7%) and significantly improved motor deficits (53.9%), circling behavior (38.9%), and long-term memory compared with ischemia/reperfusion (IR) animals. SAC increased the proportion of Ki67-positive cells (4.3-fold in the cortex and 1.8-fold in the striatum) and enhanced neurotrophin levels, NGF (cortex), BDNF (cortex and striatum), VEGF (striatum), pTrkB, pAKT, and pERK (cortex and striatum). Conclusions: SAC supports post-ischemic recovery, improving motor performance and preserving long-term recognition memory, effects that could be associated with increased cell proliferation, neurotrophin levels, and activation of the TrkB, AKT, and ERK pathways.

Background: Human dental pulp stem cells (hDPSCs) are a promising source of multipotent mesenchymal stem cells (MSCs) for regenerative neurology because of their inherent neurogenic potential. However, robust and reproducible protocols for driving their terminal neuronal maturation in a fully defined, xeno-free environment are lacking. Methods: hDPSCs were isolated from a donor tooth and characterized for mesenchymal (CD105, CD90, CD73, CD13) and stemness-associated markers (SOX2, Oct3/4 and Nanog). Cells were differentiated in a novel, fully chemically defined medium 1% ITS medium (ITS: Insulin, Transferrin, Selenium) supplemented with glial cell line-derived neurotrophic factor (GDNF) or brain-derived neurotrophic factor (BDNF). Neuronal commitment and partial maturation were assessed via immunofluorescence, Western blot, and RT-PCR for markers such as NeuN (Neuronal nuclei) and NF-M (Neurofilament medium chain), and functionally by whole-cell patch-clamp electrophysiology. Results: Although undifferentiated hDPSCs expressed neural progenitor markers (βIII-tubulin and Nestin), only GDNF treatment in a chemically defined medium significantly upregulated mature neuronal markers (NeuN and NF-M) and downregulated mesenchymal markers. Importantly, GDNF-treated cells exhibited key functional changes, including hyperpolarized resting membrane potentials, increased membrane capacitance, and elevated input resistance, which are electrophysiological hallmarks of neural precursor or early neuronal maturation, compared to control cells cultured in medium containing fetal bovine serum (FBS). Although action potentials were not elicited, this represents a significant advancement toward achieving a functional neuronal state. Conclusion: This study demonstrates that a fully chemically defined medium enables GDNF to drive hDPSCs beyond the neural progenitor state towards a partially mature neuronal phenotype. This defined medium protocol eliminates serum variability, enhances reproducibility, and provides a critical step towards standardizing hDPSC-derived neuronal cells for disease modeling and cell-based therapy.

Background: In neurodegenerative diseases such as tauopathies, including Alzheimer's disease dementia (AD) and progressive supranuclear palsy (PSP), cellular processes such as survival, proliferation, and cellular communication are highly affected. These alterations are linked to changes in the expression of various trophic factors. Various cellular models are employed to study certain tauopathies, including peripheral cells. An innovative model involves human olfactory neuroepithelial progenitor cells (hONE‐NPCs). Therefore, analyzing and characterizing the trophic factors released by hONE‐NPC‐AD and hONE‐NPC‐PSP could provide insights into the alterations occurring during the progression of these diseases (Lampinen et al., 2022). Method: Through immunodetection, cells from Mexican healthy controls from two age groups (Age 30‐40 and 50‐60 years old) and Mexican patients with Alzheimer's disease dementia and progressive supranuclear palsy were characterized. Conditioned media were first purified and concentrated using centrifugation and acetone precipitation. Subsequently, electrophoresis and Coomassie blue staining were performed to identify trophic factors present in the conditioned media through mass spectrometry. Result: hONE‐NPCs expressed proteins such as Ki67, Nestin, GFAP, and b‐III Tubulin, which are associated with cellular proliferation and the neuronal progenitor lineage. The functional characterization of hONE‐NPCs in AD and PSP patients revealed duplication times of approximately 90 hours. In the conditioned media, a differential expression of trophic factors released by hONE‐NPCs from AD and PSP patients was observed compared to apparently healthy age‐matched controls and young controls. Conclusion: The differential expression of trophic factors released by hONE‐NPCs in AD and PSP patients is significant compared to apparently healthy participants. These trophic factors are linked to signaling pathways involved in cellular proliferation and survival. This reduction directly impacts cellular proliferation and survival, as demonstrated by phenotypic characterization, showing decreased Ki67 expression relative to the total number of DAPI‐stained cells. To date, no studies have focused on the conditioned media or the changes in trophic factors during tauopathies, such as Alzheimer's disease dementia and PSP, in primary cultures of human olfactory neuroepithelial progenitor cells, opening a new research avenue.