Green synthesis of chitooligosaccharide-PEGDA derivatives through aza-Michael reaction for biomedical applications.

Chitooligosaccharide (COS) as an emerging material carbohydrate polymer with huge potential in biomedical applications was prepared using a microwave-assisted process. The obtained COS exhibited reduced molecular weight (Mw) and higher water solubility in comparison to chitosan while preserving the main saccharide structure and same degree of deacetylation (DD). The optimized COS (13 kDa) was then used to synthesize a new family of COS-poly(ethylene glycol) diacrylate (PEGDA) derivatives based on aza-Michael addition of acrylate groups of PEGDA to the amine groups of COS in the absence of any exterior agents. The modulation of the reaction time, temperature, pH and NH2:acrylate molar ratio, had a strong influence on the Michael reaction progress. At higher degrees of conversion of acrylate groups, COS-PEGDA derivative formed gel with high biocompatibility towards human bone mesenchymal stem cells (hBMSCs). These COS-PEGDA hydrogels synthesized at mild conditions through a green chemistry are, therefore, an innovative system combining adequate biological performance, ease of preparation, and an environmentally friendly concept of production.

Small Oligonucleotides Detection in Three-Dimensional Polymer Network of DNA-PEG Hydrogels.

The control of the three-dimensional (3D) polymer network structure is important for permselective materials when specific biomolecule detection is needed. Here we investigate conditions to obtain a tailored hydrogel network that combines both molecular filtering and molecular capture capabilities for biosensing applications. Along this line, short oligonucleotide detection in a displacement assay is set within PEGDA hydrogels synthetized by UV radical photopolymerization. To provide insights on the molecular filter capability, diffusion studies of several probes (sulforhodamine G and dextrans) with different hydrodynamic radii were carried out using NMR technique. Moreover, fluorometric analyses of hybridization of DNA oligonucleotides inside PEGDA hydrogels shed light on the mechanisms of recognition in 3D, highlighting that mesh size and crowding effect greatly impact the hybridization mechanism on a polymer network. Finally, we found the best probe density and diffusion transport conditions to allow the specific oligonucleotide capture and detection inside PEGDA hydrogels for oligonucleotide detection and the filtering out of higher molecular weight molecules.

Photoencapsulated-BMP2 in visible light-cured thiol-acrylate hydrogels for craniofacial bone tissue engineering.

Aim: The study aimed to examine the impact of crosslinking BMP2 in biodegradable visible light-cured thiol-acrylate hydrogels. Materials & methods: BMP2 was photoencapsulated in 10 wt% PEG-diacrylate hydrogels with or without immortalized mouse bone marrow stromal cells (BMSC). Results & conclusion: Photoencapsulated-BMSC with BMP2 (BMBMP2) showed a significantly (p < 0.05) increased level in metabolic activity, by 54.61%, compared with photoencapsulated-BMSC at day 3. Furthermore, BMBMP2 groups showed significantly increased levels in ALP activity compared with BMSC at days, 1, 3, 7 (p < 0.01) and 10 (p < 0.05). This study shows promising results photoencapsulating BMP2 in thiol-acrylate hydrogels for craniofacial bone tissue engineering applications.

Photoencapsulated-mesenchymal stromal cells in biodegradable thiol-acrylate hydrogels enhance regeneration of craniofacial bone tissue defects.

Aim: This study investigated biodegradable thiol-acrylate hydrogels as stem cell carriers to facilitate cranial bone regeneration. Materials & methods: Two formulations of thiol-acrylate hydrogels (5 and 15 wt% Poly[ethylene glycol]-diacrylate [PEGDA] hydrogels) were used as stem cell carriers. Bone marrow mesenchymal stromal cells and dental pulp mesenchymal stromal cells were photoencapsulated and cultured in basal or osteogenic medium 3 days before the surgery. Using New Zealand White Rabbits, four defects (5 mm diameter and 2 mm thickness) were created and hydrogel scaffolds were implanted in each rabbit cranium for 6 weeks. Results & Conclusion: AlamarBlue assay showed increasing metabolic activity levels in 5 wt% PEGDA hydrogels than 15 wt% PEGDA hydrogels. Photoencapsulated-mesenchymal stromal cells in 15 wt% PEGDA hydrogels demonstrated significantly increasing alkaline phosphatase activity levels on day 7 compared with days 1 and 3. Histological diagnosis showed 5 wt% PEGDA hydrogels resulted in lower averaged residual gel areas than 15 wt% PEGDA hydrogel specimens and control groups 6 weeks postimplantation.

Evaluating Osteogenic Potential of Ligamentum Flavum Cells Cultivated in Photoresponsive Hydrogel that Incorporates Bone Morphogenetic Protein-2 for Spinal Fusion.

Regenerative medicine is increasingly important in clinical practice. Ligamentum flava (LF) are typically removed during spine-related surgeries. LF may be a source of cells for spinal fusion that is conducted using tissue engineering techniques. In this investigation, LF cells of rabbits were isolated and then characterized by flow cytometry, morphological observation, and immunofluorescence staining. The LF cells were also cultivated in polyethylene (glycol) diacrylate (PEGDA) hydrogels that incorporated bone morphogenetic protein-2 (BMP-2) growth factor, to evaluate their proliferation and secretion of ECM and differentiation in vitro. The experimental results thus obtained that the proliferation, ECM secretion, and differentiation of the PEGDA-BMP-2 group exceeded those of the PEGDA group during the period of cultivation. The mineralization and histological staining results differed similarly. A nude mice model was utilized to prove that LF cells on hydrogels could undergo osteogenic differentiation in vivo. These experimental results also revealed that the PEGDA-BMP-2 group had better osteogenic effects than the PEGDA group following a 12 weeks after transplantation. According to all of these experimental results, LF cells are a source of cells for spinal fusion and PEGDA-BMP-2 hydrogel is a candidate biomaterial for spinal fusion by tissue engineering.