Assessment of Cytocompatibility and AntiInflammatory (Inter)Actions of Genipin-Crosslinked Chitosan Powders

none5Chitosan is a polysaccharide commonly used, together with its derivatives, in the preparation of hydrogel formulations, scaffolds and films for tissue engineering applications. Chitosan can be used as such, but it is commonly stabilized by means of chemical crosslinkers. Genipin is one of the crosslinkers that has been considered that is a crystalline powder extracted from the fruit of Gardenia jasminoides and processed to obtain an aglycon compound. Genipin is gaining interest in biological applications because of its natural origin and anti-inflammatory actions. In this paper, the ability of chitosan-based materials crosslinked with genipin to exert anti-inflammation properties in applications such as bone regeneration was studied. Powders obtained from chitosan–genipin scaffolds have been tested in order to mimic the natural degradation processes occurring during biomaterials implantation in vivo. The results from osteoblast-like cells showed that specific combinations of chitosan and genipin stimulate high permissiveness towards cells, with higher performance than the pure chitosan. In parallel, evidences from monocyte-like cells showed that the crosslinker, genipin, seems to promote slowing of the monocyte-macrophage transition at morphological level. This suggests a sort of modularity of pro-inflammatory versus anti-inflammatory behavior of our chitosan-based biomaterials. Being both the cell types exposed to microscale powders, as an added value our results bring information on the cell–material interactions in the degradative dynamics of chitosan scaffold structures during the physiological resorption processes.noneSimona Dimida; Matteo Santin; Tiziano Verri; Amilcare Barca; Christian DemitriDimida, Simona; Santin, Matteo; Verri, Tiziano; Barca, Amilcare; Demitri, Christia

Fifteen years of iodine prophylaxis in Italy. Results of a nationwide surveillance (Period 2015-2019)

CONTEXT: In 2005, a nationwide program of iodine prophylaxis on a voluntary basis was implemented in Italy by law. However, recent data on iodine status are lacking. OBJECTIVE: The aim of this study was to evaluate efficiency, effectiveness, and possible adverse effects (increased occurrence of thyroid autoimmunity and hyperthyroidism) of the Italian iodine prophylaxis program. METHODS: From 2015 to 2019, a nationwide survey was performed. The use of iodized salt was evaluated in a sample of 164 593 adults and in 998 school canteens. A sample of 4233 schoolchildren (aged 11-13 years) was recruited to assess urinary iodine concentration, prevalence of goiter, and thyroid hypoechogenicity on ultrasound, with the latter being an indirect indicator of thyroid autoimmunity. Neonatal TSH values of 197 677 infants screened in regions representative of Northern, Central, and Southern Italy were analyzed to investigate the percentage of TSH values >5.0 mIU/L. Data on methimazole prescriptions were analyzed as indirect indicators of new cases of hyperthyroidism. RESULTS: The prevalence of the use of iodized salt was 71.5% in adult population and 78% in school canteens. A median urinary iodine concentration of 124 μg/L, a prevalence of goiter of 2.2%, and a prevalence of thyroid hypoechogenicity of 5.7% were observed in schoolchildren. The percentage of neonatal TSH values >5.0 mIU/L resulted still higher (5.1%) than the World Health Organization threshold of 3.0%, whereas the prescriptions of methimazole showed a reduction of 13.5%. CONCLUSION: Fifteen years of iodine prophylaxis have led to iodine sufficiency in Italy, although there still is concern about iodine nutritional status during pregnancy

Assessment of Cytocompatibility and Anti-Inflammatory (Inter)Actions of Genipin-Crosslinked Chitosan Powders

Chitosan is a polysaccharide commonly used, together with its derivatives, in the preparation of hydrogel formulations, scaffolds and films for tissue engineering applications. Chitosan can be used as such, but it is commonly stabilized by means of chemical crosslinkers. Genipin is one of the crosslinkers that has been considered that is a crystalline powder extracted from the fruit of Gardenia jasminoides and processed to obtain an aglycon compound. Genipin is gaining interest in biological applications because of its natural origin and anti-inflammatory actions. In this paper, the ability of chitosan-based materials crosslinked with genipin to exert anti-inflammation properties in applications such as bone regeneration was studied. Powders obtained from chitosan–genipin scaffolds have been tested in order to mimic the natural degradation processes occurring during biomaterials implantation in vivo. The results from osteoblast-like cells showed that specific combinations of chitosan and genipin stimulate high permissiveness towards cells, with higher performance than the pure chitosan. In parallel, evidences from monocyte-like cells showed that the crosslinker, genipin, seems to promote slowing of the monocyte-macrophage transition at morphological level. This suggests a sort of modularity of pro-inflammatory versus anti-inflammatory behavior of our chitosan-based biomaterials. Being both the cell types exposed to microscale powders, as an added value our results bring information on the cell–material interactions in the degradative dynamics of chitosan scaffold structures during the physiological resorption processes.</jats:p

Study on the degradation of chitosan slurries

AbstractIn the present work, we measured the degradation rate of different chitosan slurries. Several parameters were monitored such as temperature (25°C, 37°C, 50°C); chitosan concentration (1% and 2% (w/V)); and polymer molecular weight. The samples were tested in dynamic sweep test mode. This test is able to provide a reliable estimation of viscosity variations of the slurries; in turn, these variations could be related to degradation rate of the system in the considered conditions. The resulting information is particularly important especially in applications in which there is a close relationship between physical properties and molecular structure

Development of a Novel Hybrid Porous Scaffold for Bone Tissue Engineering: Forsterite Nanopowder Reinforced Chitosan

In order to induce bone regeneration several natural and synthetic materials have been proposed. However, single-phase scaffolds present some insurmountable disadvantages such as poor mechanical strength or brittleness and too low or too high degradation rate. In order to overcome these drawbacks, composite systems can be an interesting and promising option. In the present work a novel hybrid porous scaffold for bone tissue engineering is proposed. Chitosan/Forsterite (Ch/FS) composite scaffolds were prepared by freeze-drying method using a chitosan/forsterite ratio of 90/10. The FS nanopowder (Mg2SiO4) is synthesized using a simple solgel based method. The FS composition was checked by XRD analysis. The macrostructure of the Ch/FS scaffolds were analyzed by SEM, the FS distribution within the chitosan matrix observed by EDS, the mechanical strength measured by compression test in PBS and the biocompatibility of the composite on human osteosarcoma cell line (MG-63) verified by MTT assay after 48 hours. The porosity appears interconnected and with a pore size ranging from 1 to 100 μm. The FS is overall distributed within the chitosan matrix. The compression strength of composite scaffolds increased with respect to the pure chitosan scaffolds of more than two times (from 0.8 to 1.9 KPa) and the composites did not show any toxicity effect on human osteosarcoma cells.</jats:p

Genipin-cross-linked chitosan-based hydrogels: Reaction kinetics and structure-related characteristics

This book presents a new approach to studying the syntax of human language, one which emphasizes how we think about time. Tilsen argues that many current theories are unsatisfactory because those theories conceptualize syntactic patterns with spatially arranged structures of objects. These object-structures are atemporal and do not lend well to reasoning about time. The book develops an alternative conceptual model in which oscillatory systems of various types interact with each other through coupling forces, and in which the relative energies of those systems are organized in particular ways. Tilsen emphasizes that the two primary mechanisms of the approach&nbsp;– oscillators and energy levels&nbsp;– require alternative ways of thinking about time. Furthermore, his theory leads to a new way of thinking about grammaticality and the recursive nature of language. The theory is applied to a variety of syntactic phenomena: word order, phrase structure, morphosyntax, constituency, case systems, ellipsis, anaphora, and islands. The book also presents a general program for the study of language in which the construction of linguistic theories is itself an object of theoretical analysis. Reviewed by John Goldsmith, Mark Gibson and an anonymous reviewer. Signed reports are openly available in the downloads session

Effects of Genipin Concentration on Cross-Linked Chitosan Scaffolds for Bone Tissue Engineering: Structural Characterization and Evidence of Biocompatibility Features

Genipin (GN) is a natural molecule extracted from the fruit of Gardenia jasminoides Ellis according to modern microbiological processes. Genipin is considered as a favorable cross-linking agent due to its low cytotoxicity compared to widely used cross-linkers; it cross-links compounds with primary amine groups such as proteins, collagen, and chitosan. Chitosan is a biocompatible polymer that is currently studied in bone tissue engineering for its capacity to promote growth and mineral-rich matrix deposition by osteoblasts in culture. In this work, two genipin cross-linked chitosan scaffolds for bone repair and regeneration were prepared with different GN concentrations, and their chemical, physical, and biological properties were explored. Scanning electron microscopy and mechanical tests revealed that nonremarkable changes in morphology, porosity, and mechanical strength of scaffolds are induced by increasing the cross-linking degree. Also, the degradation rate was shown to decrease while increasing the cross-linking degree, with the high cross-linking density of the scaffold disabling the hydrolysis activity. Finally, basic biocompatibility was investigated in vitro, by evaluating proliferation of two human-derived cell lines, namely, the MG63 (human immortalized osteosarcoma) and the hMSCs (human mesenchymal stem cells), as suitable cell models for bone tissue engineering applications of biomaterials

Identification of a novel pax8 gene sequence variant in four members of the same family: from congenital hypothyroidism with thyroid hypoplasia to mild subclinical hypothyroidism.

Abstract Background: Congenital hypothyroidism is often secondary to thyroid dysgenesis, including thyroid agenesis, hypoplasia, ectopic thyroid tissue or cysts. Loss of function mutations in TSHR, PAX8, NKX2.1, NKX2.5 and FOXE1 genes are responsible for some forms of inherited congenital hypothyroidism, with or without hypoplastic thyroid. The aim of this study was to analyse the PAX8 gene sequence in several members of the same family in order to understand whether the variable phenotypic expression, ranging from congenital hypothyroidism with thyroid hypoplasia to mild subclinical hypothyroidism, could be associated to the genetic variant in the PAX8 gene, detected in the proband. Methods: We screened a hypothyroid child with thyroid hypoplasia for mutations in PAX8, TSHR, NKX2.1, NKX2.5 and FOXE1 genes. We studied the inheritance of the new variant R133W detected in the PAX8 gene in the proband’s family, and we looked for the same substitution in 115 Caucasian European subjects and in 26 hypothyroid children. Functional studies were performed to assess the in vitro effect of the newly identified PAX8 gene variant. Results: A new heterozygous nucleotide substitution was detected in the PAX8 DNA-binding motif (c.397C/T, R133W) in the proband, affected by congenital hypothyroidism with thyroid hypoplasia, in his older sister, displaying a subclinical hypothyroidism associated with thyroid hypoplasia and thyroid nodules, in his father, affected by hypothyroidism with thyroid hypoplasia and thyroid nodules, and his first cousin as well, who revealed only a subclinical hypothyroidism. Functional studies of R133W-PAX8 in the HEK293 cells showed activation of the TG promoter comparable to the wild-type PAX8. Conclusions: In vitro data do not prove that R133W-PAX8 is directly involved in the development of the thyroid phenotypes reported for family members carrying the substitution. However, it is reasonable to conceive that, in the cases of transcriptions factors, such as Pax8, which establish several interactions in different protein complexes, genetic variants could have an impact in vivo. Keywords: PAX8 gene, Thyroid, Congenital hypothyroidism, Variable phenotypic expressivity, R133W-PAX