Dissociation between two aspects of procedural learning in Tourette syndrome: Enhanced statistical and impaired sequence learning.

Tourette syndrome (TS) is a childhood-onset neurodevelopmental disorder that primarily affects the cortico-basal ganglia-thalamo-cortical (CBGTC) circuitry and is characterized by motor and vocal tics. Previous studies have found enhancement in procedural memory, which depends on the CBGTC circuitry and plays an important role in the learning and processing of numerous motor, social, and cognitive skills and habits. Based on these studies, procedural hyperfunctioning in TS has been proposed. However, the neurocognitive mechanism underlying such hyperfunctioning is poorly understood. Here, we investigated how two aspects of procedural learning, namely 1) frequency-based statistical learning and 2) order-based sequence learning, are affected in TS. Twenty-one children with TS between the ages of ten and fifteen as well as 21 typically developing controls were tested on a probabilistic sequence learning task that enables the parallel assessment of these two aspects. We found that children with TS showed enhanced sensitivity to statistical information but impaired sequence learning compared to typically developing children. The deconstruction of procedural memory suggests that procedural hyperfunctioning in TS may be supported by enhanced sensitivity to statistical information. These results can provide a potential path for improving therapy methods and skill-oriented educational programs for TS.

STRO-1 positive cell expansion during osteogenic differentiation: A comparative study of three mesenchymal stem cell types of dental origin.

OBJECTIVE: Although the osteogenic differentiation potential of mesenchymal stem cells of dental origin is well established, the roles of different marker proteins in this process remain to be clarified. Our aim was to compare the cellular and molecular changes, focusing in particular on mesenchymal stem cell markers, during in vitro osteogenesis in three dental stem cell types: dental follicle stem cells (DFSCs), periodontal ligament stem cells (PDLSCs) and dental pulp stem cells (DPSCs). DESIGN: Human DFSCs, PDLSCs and DPSCs were isolated, cultured and their osteogenic differentiation was induced for 3 weeks. Mineralization was assessed by von Kossa staining and calcium concentration measurements. The expression of mesenchymal and osteogenic markers was studied by immunocytochemistry and qPCR techniques. Alkaline phosphatase (ALP) activity and the frequency of STRO-1 positive cells were also quantified. RESULTS: The three cultures all showed abundant mineralization, with high calcium content by day 21. The expression of vimentin and nestin was sustained after osteogenic induction. The osteogenic medium induced a considerable elevation of STRO-1 positive cells. By day 7, the ALP mRNA level had increased more than 100-fold in DFSCs, PDLSCs, and DPSCs. Quantitative PCR results indicated dissimilarities of osteoblastic marker levels in the three dental stem cell cultures. CONCLUSIONS: DFSCs, PDLSCs and DPSCs have similar functional osteogenic differentiation capacities although their expressional profiles of key osteogenic markers show considerable variations. The STRO-1 positive cell fraction expands during osteogenic differentiation while vimentin and nestin expression remain high. For identification of stemness, functional studies rather than marker expressions are needed.

Maintained Visual-, Auditory-, and Multisensory-Guided Associative Learning Functions in Children With Obsessive-Compulsive Disorder.

Sensory-guided acquired equivalence learning, a specific kind of non-verbal associative learning, is associated with the frontal cortex-basal ganglia loops and hippocampi, which seem to be involved in the pathogenesis of obsessive-compulsive disorder (OCD). In this study, we asked whether visual-, auditory-, and multisensory-guided associative acquired equivalence learning is affected in children with OCD. The first part of the applied learning paradigm investigated association building between two different sensory stimuli (where feedback was given about the correctness of the choices), a task that critically depends upon the basal ganglia. During the test phases, which primarily depended upon the hippocampi, the earlier learned and hitherto not shown but predictable associations were asked about without feedback. This study involved 31 children diagnosed with OCD according to the Diagnostic and Statistical Manual of Mental Disorders, 5th Edition (DSM-V) criteria and 31 matched healthy control participants. The children suffering from OCD had the same performance as the control children in all phases of the applied visual-, auditory-, and multisensory-guided associative learning paradigms. Thus, both the acquisition and test phases were not negatively affected by OCD. The reaction times did not differ between the two groups, and the applied medication had no effect on the performances of the OCD patients. Our results support the findings that the structural changes of basal ganglia and hippocampi detected in adult OCD patients are not as pronounced in children, which could be the explanation of the maintained associative equivalence learning functions in children suffering from OCD.

Impairment of visually guided associative learning in children with Tourette syndrome.

The major symptoms of Tourette syndrome are motor and vocal tics, but Tourette syndrome is occasionally associated with cognitive alterations as well. Although Tourette syndrome does not affect the majority of cognitive functions, some of them improve. There is scarce evidence on the impairment of learning functions in patients with Tourette syndrome. The core symptoms of Tourette syndrome are related to dysfunction of the basal ganglia and the frontostriatal loops. Acquired equivalence learning is a kind of associative learning that is related to the basal ganglia and the hippocampi. The modified Rutgers Acquired Equivalence Test was used in the present study to observe the associative learning function of patients with Tourette syndrome. The cognitive learning task can be divided into two main phases: the acquisition and test phases. The latter is further divided into two parts: retrieval and generalization. The acquisition phase of the associative learning test, which mainly depends on the function of the basal ganglia, was affected in the entire patient group, which included patients with Tourette syndrome with attention deficit hyperactivity disorder, obsessive compulsive disorder, autism spectrum disorder, or no comorbidities. Patients with Tourette syndrome performed worse in building associations. However, the retrieval and generalization parts of the test phase, which primarily depend on the function of the hippocampus, were not worsened by Tourette syndrome.

Free thiol groups on poly(aspartamide) based hydrogels facilitate tooth-derived progenitor cell proliferation and differentiation.

Cell-based tissue reconstruction is an important field of regenerative medicine. Stem and progenitor cells derived from tooth-associated tissues have strong regeneration potential. However, their in vivo application requires the development of novel scaffolds that will provide a suitable three-dimensional (3D) environment allowing not only the survival of the cells but eliciting their proliferation and differentiation. Our aim was to study the viability and differentiation capacity of periodontal ligament cells (PDLCs) cultured on recently developed biocompatible and biodegradable poly(aspartamide) (PASP)-based hydrogels. Viability and behavior of PDLCs were investigated on PASP-based hydrogels possessing different chemical, physical and mechanical properties. Based on our previous results, the effect of thiol group density in the polymer matrix on cell viability, morphology and differentiation ability is in the focus of our article. The chemical composition and 3D structures of the hydrogels were determined by FT Raman spectroscopy and Scanning Electron Microscopy. Morphology of the cells was examined by phase contrast microscopy. To visualize cell growth and migration patterns through the hydrogels, two-photon microscopy were utilized. Cell viability analysis was performed according to a standardized protocol using WST-1 reagent. PDLCs were able to attach and grow on PASP-based hydrogels. An increase in gel stiffness enhanced adhesion and proliferation of the cells. However, the highest population of viable cells was observed on the PASP gels containing free thiol groups. The presence of thiol groups does not only enhance viability but also facilitates the osteogenic direction of the differentiating cells. These cell-gel structures seem to be highly promising for cell-based tissue reconstruction purposes in the field of regenerative medicine.

Fully amino acid-based hydrogel as potential scaffold for cell culturing and drug delivery.

Polymer hydrogels are ideal scaffolds for both tissue engineering and drug delivery. A great advantage of poly(amino acid)-based hydrogels is their high similarity to natural proteins. However, their expensive and complicated synthesis often limits their application. The use of poly(aspartic acid) (PASP) seems an appropriate solution for this problem due to the relatively cheap and simple synthesis of PASP. Using amino acids not only as building blocks in the polymer backbone but also as cross-linkers can improve the biocompatibility and the biodegradability of the hydrogel. In this paper, PASP cross-linked with cystamine (CYS) and lysine-methylester (LYS) was introduced as fully amino acid-based polymer hydrogel. Gels were synthesized employing six different ratios of CYS and LYS. The pH dependent swelling degree and the concentration of the elastically active chain were determined. After reduction of the disulfide bonds of CYS, the presence of thiol side groups was also detected. To determine the concentration of the reactive cross-linkers in the hydrogels, a new method based on the examination of the swelling behavior was established. Using metoprolol as a model drug, cell proliferation and drug release kinetics were studied at different LYS contents and in the presence of thiol groups. The optimal ratio of cross-linkers for the proliferation of periodontal ligament cells was found to be 60-80% LYS and 20-40% CYS. The reductive conditions resulted in an increased drug release due to the cleavage of disulfide bridges in the hydrogels. Consequently, these hydrogels provide new possibilities in the fields of both tissue engineering and controlled drug delivery.

Effect of stem cells of dental pulp origin on osseointegration of titanium implant in a novel rat vertebra model.

During that last decade a large number of experiments showed the successful application of stem cells in achieving large bone volume regeneration. On the contrary, our knowledge about the promotion of implant osseointegration by stem cell is sporadic. Recently, our research group has carried out an array of studies aiming the characterization of postnatal stem cells of dental origin. In addition, we have developed a novel quantitative model for implant osseointegration in rat tail vertebrae. In the present work we aimed to study how the implant osseointegration process is affected by mesenchymal stem cells of rat dental pulp origin (DPSC) when cells are undifferentiated or predifferentiated into osteogenic direction. Our results show that undifferentiated pulp cells inserted between the implant and the bone slow down the osseointegration process. On the other hand, pre-differentiated DPSCs do not have a similar adverse effect any more. Our data suggest that the success of mesenchymal stem cell application to promote implant osseointegration is highly dependent on the applied conditions, particularly on the parallel applicatioh of scaffolds and osteogenic components.