RESUMEN
Amelogenins are enamel matrix proteins currently used to treat bone defects in periodontal surgery. Recent studies have highlighted the relevance of amelogenin-derived peptides, named LRAP, TRAP, SP, and C11, in bone tissue engineering. Interestingly, these peptides seem to maintain or even improve the biological activity of the full-length protein, which has received attention in the field of bone regeneration. In this article, the authors combined a systematic and a narrative review. The former is focused on the existing scientific evidence on LRAP, TRAP, SP, and C11's ability to induce the production of mineralized extracellular matrix, while the latter is concentrated on the structure and function of amelogenin and amelogenin-derived peptides. Overall, the collected data suggest that LRAP and SP are able to induce stromal stem cell differentiation towards osteoblastic phenotypes; specifically, SP seems to be more reliable in bone regenerative approaches due to its osteoinduction and the absence of immunogenicity. However, even if some evidence is convincing, the limited number of studies and the scarcity of in vivo studies force us to wait for further investigations before drawing a solid final statement on the real potential of amelogenin-derived peptides in bone tissue engineering.
Asunto(s)
Amelogenina/metabolismo , Regeneración Ósea/fisiología , Péptidos/metabolismo , Amelogenina/química , Amelogenina/genética , Secuencia de Aminoácidos , Animales , Biomarcadores , Diferenciación Celular , Regulación de la Expresión Génica , Humanos , Inmunohistoquímica , Péptidos/química , Ingeniería de Tejidos , Investigación Biomédica TraslacionalRESUMEN
Matrix metalloproteinase-2 (MMP-2) is an endopeptidase involved in cardiovascular disease and cancer. To date, no highly selective MMP-2 inhibitors have been identified for potential use in humans. Aim of our work was to apply the nanobody technology to the generation of highly selective inhibitors of human MMP-2 and to assess their effects on platelet function and their applicability as conjugated nanobodies. We constructed a nanobody library after immunising an alpaca with human active MMP-2 and identified, after phage display and screening, one MMP-2 inhibitory nanobody (VHH-29), able to hinder the effects of MMP-2 on platelet activation, and one nanobody not inhibiting MMP-2 activity (VHH-136) which, chemically conjugated to a fluorescent probe, allowed the detection of human MMP-2 by flow-cytometry and immune-cytochemistry. In conclusion, we have generated and characterized two new nanotechnological molecular tools for human MMP-2 which represent promising agents for the study of MMP-2 in cardiovascular pathophysiology.
Asunto(s)
Citometría de Flujo , Metaloproteinasa 2 de la Matriz/inmunología , Biblioteca de Péptidos , Anticuerpos de Dominio Único , Ensayo de Inmunoadsorción Enzimática , Células HEK293 , Humanos , Anticuerpos de Dominio Único/química , Anticuerpos de Dominio Único/inmunologíaAsunto(s)
Mutación con Ganancia de Función , Genes Dominantes , Integrina beta3/genética , Mutación con Pérdida de Función , Trombastenia/diagnóstico , Trombastenia/genética , Adulto , Alelos , Sustitución de Aminoácidos , Biomarcadores , Plaquetas/metabolismo , Plaquetas/ultraestructura , Análisis Mutacional de ADN , Índices de Eritrocitos , Femenino , Estudios de Asociación Genética , Humanos , Linaje , Fenotipo , Pruebas de Función PlaquetariaRESUMEN
BACKGROUND: The GeneXpert analyzer is a hands-off system for the detection of Factor V Leiden and of Prothrombin G20210A (GPRO) gene thrombophilic mutations. Although the system is efficient and easy to use, we report the rare possibility of incorrect genotyping. METHODS: 1648 samples were evaluated using the GeneXpert HemosIL Factor II and Factor V assay: 1319 were freshly analyzed while 329 were frozen, thawed and diluted with saline prior to analysis to avoid clogging of the instrument syringe. RESULTS: Two samples, both heterozygous, one for the factor V Leiden and the other for the GPRO gene, were incorrectly genotyped as homozygous for the relative mutation. Inspection of the Ct values and amplification curves and genotyping with PCR revealed the correct genotype as heterozygous for factor V Leiden and GPRO mutation. DISCUSSION: The GeneXpert HemosIL Factor II and Factor V assay is an automated, fast genotyping assay requiring almost no sample manipulation, advantageous characteristics if compared with other PCR-based methods. However, an inattentive use of it can generate incorrect diagnosis. A careful handling of the sample, in particular correct dilution of frozen/thawed samples before analysis, and the inspection of the amplification curves and Ct values are required to avoid artifacts.
