Three-dimensional printing and in vitro evaluation of poly(3-hydroxybutyrate) scaffolds functionalized with osteogenic growth peptide for tissue engineering.

Poly(3-hydroxybutyrate) (PHB) is a biodegradable and thermoprocessable biopolymer, making it a promising candidate for applications in tissue engineering. In the present study a structural characterization and in vitro evaluation were performed on PHB scaffolds produced by additive manufacturing via selective laser sintering (SLS), followed by post-printing functionalization with osteogenic growth peptide (OGP) and its C-terminal sequence OGP(10-14). The PHB scaffolds were characterized, including their morphology, porosity, thermal and mechanical properties, moreover OGP release. The results showed that SLS technology allowed the sintering of the PHB scaffolds with a hierarchical structure with interconnected pores and intrinsic porosity (porosity of 55.8 ±â€¯0.7% and pore size in the 500-700 µm range), and good mechanical properties. Furthermore, the SLS technology did not change thermal properties of PHB polymer. The OGP release profile showed that PHB scaffold promoted a controlled release above 72 h. In vitro assays using rat bone marrow stem cells showed good cell viability/proliferation in all the PHB scaffolds. Additionally, SEM images suggested advanced morphological differentiation in the groups containing osteogenic growth peptide. The overall results demonstrated that PHB biopolymer is potential candidate for 3D printing via SLS technology, moreover the OGP-containing PHB scaffolds showed ability to sustain cell growth to support tissue formation thereby might be considered for tissue-engineering applications.

Evaluation of genetic variation among Brazilian soybean cultivars through genome resequencing.

BACKGROUND: Soybean [Glycine max (L.) Merrill] is one of the most important legumes cultivated worldwide, and Brazil is one of the main producers of this crop. Since the sequencing of its reference genome, interest in structural and allelic variations of cultivated and wild soybean germplasm has grown. To investigate the genetics of the Brazilian soybean germplasm, we selected soybean cultivars based on the year of commercialization, geographical region and maturity group and resequenced their genomes. RESULTS: We resequenced the genomes of 28 Brazilian soybean cultivars with an average genome coverage of 14.8X. A total of 5,835,185 single nucleotide polymorphisms (SNPs) and 1,329,844 InDels were identified across the 20 soybean chromosomes, with 541,762 SNPs, 98,922 InDels and 1,093 CNVs that were exclusive to the 28 Brazilian cultivars. In addition, 668 allelic variations of 327 genes were shared among all of the Brazilian cultivars, including genes related to DNA-dependent transcription-elongation, photosynthesis, ATP synthesis-coupled electron transport, cellular respiration, and precursors of metabolite generation and energy. A very homogeneous structure was also observed for the Brazilian soybean germplasm, and we observed 41 regions putatively influenced by positive selection. Finally, we detected 3,880 regions with copy-number variations (CNVs) that could help to explain the divergence among the accessions evaluated. CONCLUSIONS: The large number of allelic and structural variations identified in this study can be used in marker-assisted selection programs to detect unique SNPs for cultivar fingerprinting. The results presented here suggest that despite the diversification of modern Brazilian cultivars, the soybean germplasm remains very narrow because of the large number of genome regions that exhibit low diversity. These results emphasize the need to introduce new alleles to increase the genetic diversity of the Brazilian germplasm.