Engineering bioactive 3D printing bioinks towards targeted personalised therapies

Constructing functional scaffolds for targeted controlled drug release and tissue regeneration offers promising therapies for many diseases such as tumours, bone regeneration, wound healing, bacterial and fungal infections. 3D bioprinting is an additive manufacturing approach that utilizes a “bioink” to fabricate complex structures out of molecules, similar to assembling Lego pieces. The ideal should satisfy certain material (printability, mechanics, degradation, functionalisation) and biological requirements (biocompatibility, cytocompatilibilty, and bioactivity). Inclusion or conjugation of peptides is common within bioinks as specific regulators of cell activities or for their therapeutic potential. However, the lack of peptide stability to enzymatic degradation remains a problem in realising their potential. We have shown that peptide amphiphiles (PAs), prepared by introducing lipidic parts within peptides, are highly stable and stop the growth of brain and breast cancer cells alone or when loaded with chemotherapeutics. We aim to develop 3D printable biodegradable bioactive bio-inks prepared by cellulose based cell-friendly bioinks loaded with peptides or peptide amphiphiles that when printed into patient specific implants can elicit a specific anticancer effect and target chemotherapeutics to cancer cells remaining after surgery towards targeted and personalised treatments in cancer. In this respect, in this TRIF project, we have synthesised cellulose nanocrystals and we have used them to prepare 3D printable gels when mixed with calcium chloride. We have studied and optimised the rheological properties of cellulose nanocrystal hydrogels and characterised them in terms of particle size and morphology. Finally, we have embedded a model peptide, leucine encephalin, and studied its release from 3D printed scaffolds. We are planning to embedded an antiproliferative peptide amphiphile using similar protocols and understand its antiproliferative effects in vitro using breast and brain tumour cell lines (MCF-7, MDA-MD-231, U87MG)

États généraux du droit médical et du dommage corporel. Tome 1. Corps amputé, corps prothésé :simple puzzle de la réparation ?

info:eu-repo/semantics/publishe

Pt-flax (phenotyping and tilling of flax): development of a flax (linum usitatissimum l.) mutant population and tilling platform for forward and reverse genetics

Background: Flax (Linum usitatissimum L.) is an economically important fiber and oil crop that has been grown for thousands of years. The genome has been recently sequenced and transcriptomics are providing information on candidate genes potentially related to agronomically-important traits. In order to accelerate functional characterization of these genes we have generated a flax EMS mutant population that can be used as a TILLinG (Targeting Induced Local Lesions in Genomes) platform for forward and reverse genetics. Results: A population of 4,894 M2 mutant seed families was generated using 3 different EMS concentrations (0.3%, 0.6% and 0.75%) and used to produce M2 plants for subsequent phenotyping and DNA extraction. 10,839 viable M2 plants (4,033 families) were obtained and 1,552 families (38.5%) showed a visual developmental phenotype (stem size and diameter, plant architecture, flower-related). The majority of these families showed more than one phenotype. Mutant phenotype data are organised in a database and can be accessed and searched at UTILLdb (http://urgv.evry.inra.fr/UTILLdb). Preliminary screens were also performed for atypical fiber and seed phenotypes. Genomic DNA was extracted from 3,515 M2 families and eight-fold pooled for subsequent mutant detection by ENDO1 nuclease mis-match cleavage. In order to validate the collection for reverse genetics, DNA pools were screened for two genes coding enzymes of the lignin biosynthesis pathway: Coumarate-3-Hydroxylase (C3H) and Cinnamyl Alcohol Dehydrogenase (CAD). We identified 79 and 76 mutations in the C3H and CAD genes, respectively. The average mutation rate was calculated as 1/41 Kb giving rise to approximately 9,000 mutations per genome. Thirty-five out of the 52 flax cad mutant families containing missense or codon stop mutations showed the typical orange-brown xylem phenotype observed in CAD down-regulated/mutant plants in other species. Conclusions: We have developed a flax mutant population that can be used as an efficient forward and reverse genetics tool. The collection has an extremely high mutation rate that enables the detection of large numbers of independant mutant families by screening a comparatively low number of M2 families. The population will prove to be a valuable resource for both fundamental research and the identification of agronomically-important genes for crop improvement in flax

Pt-flax (phenotyping and tilling of flax): development of a flax (linum usitatissimum l.) mutant population and tilling platform for forward and reverse genetics

Background: Flax (Linum usitatissimum L.) is an economically important fiber and oil crop that has been grown for thousands of years. The genome has been recently sequenced and transcriptomics are providing information on candidate genes potentially related to agronomically-important traits. In order to accelerate functional characterization of these genes we have generated a flax EMS mutant population that can be used as a TILLinG (Targeting Induced Local Lesions in Genomes) platform for forward and reverse genetics. Results: A population of 4,894 M2 mutant seed families was generated using 3 different EMS concentrations (0.3%, 0.6% and 0.75%) and used to produce M2 plants for subsequent phenotyping and DNA extraction. 10,839 viable M2 plants (4,033 families) were obtained and 1,552 families (38.5%) showed a visual developmental phenotype (stem size and diameter, plant architecture, flower-related). The majority of these families showed more than one phenotype. Mutant phenotype data are organised in a database and can be accessed and searched at UTILLdb (http://urgv.evry.inra.fr/UTILLdb). Preliminary screens were also performed for atypical fiber and seed phenotypes. Genomic DNA was extracted from 3,515 M2 families and eight-fold pooled for subsequent mutant detection by ENDO1 nuclease mis-match cleavage. In order to validate the collection for reverse genetics, DNA pools were screened for two genes coding enzymes of the lignin biosynthesis pathway: Coumarate-3-Hydroxylase (C3H) and Cinnamyl Alcohol Dehydrogenase (CAD). We identified 79 and 76 mutations in the C3H and CAD genes, respectively. The average mutation rate was calculated as 1/41 Kb giving rise to approximately 9,000 mutations per genome. Thirty-five out of the 52 flax cad mutant families containing missense or codon stop mutations showed the typical orange-brown xylem phenotype observed in CAD down-regulated/mutant plants in other species. Conclusions: We have developed a flax mutant population that can be used as an efficient forward and reverse genetics tool. The collection has an extremely high mutation rate that enables the detection of large numbers of independant mutant families by screening a comparatively low number of M2 families. The population will prove to be a valuable resource for both fundamental research and the identification of agronomically-important genes for crop improvement in flax