Aminated ß-Glucan with immunostimulating activities and collagen composite sponge for wound repair.

Immunostimulating activities of yeast ß(1 â†’ 3)-D-Glucan (ß-Glucan) mainly depended on its structures. However, due to the tight triple helix structure of ß-Glucan, its immunostimulating activity is greatly weakened. Therefore, in order to partially unwind the tight triple helix structure of ß-glucan and improve its solubility in the medium, we modified it by amination in this study (A-Glu). The results showed that A-Glu could stimulate Raw264.7 macrophages and significantly enhance its TNF-α, IL-6, and IL-10 cytokine expression levels in vitro. A-Glu could also induce a shift of M0 Raw264.7 toward M1, and M2 toward M1. To expand the application of A-Glu in wound repair, the composite sponge consisting of A-Glu and type I collagen via the formation of a stable polyion complex (PIC) was developed. Moreover, the composite sponge could accelerate wound repair significantly. These results reveal that soluble A-Glu as an immunostimulating agent has potential applications in biomedicine.

Sequentially releasing self-healing hydrogel fabricated with TGFß3-microspheres and bFGF to facilitate rat alveolar bone defect repair.

Resorption and loss of alveolar bone leads to oral dysfunction and loss of natural or implant teeth. Biomimetic delivery of growth factors based on stem cell recruitment and osteogenic differentiation, as the key steps in natural alveolar bone regenerative process, has been an area of intense research in recent years. A mesoporous self-healing hydrogel (DFH) with basic fibroblast growth factor (bFGF) entrapment and transforming growth factor ß3 (TGFß3) - loaded chitosan microspheres (CMs) was developed. The formulation was optimized by multiple tests of self-healing, in-bottle inversion, SEM, rheological, swelling rate and in vitro degradation. In vitro tubule formation assays, cell migration assays, and osteogenic differentiation assays confirmed the ability of DFH to promote blood vessels, recruit stem cells, and promote osteogenic differentiation. The optimum DFH formula is 0.05 ml 4Arm-PEG-DF (20%) added to 1 ml CsGlu (2%) containing bFGF (80 ng) and TGFß3-microspheres (5 mg). The results of in vitro release studied by Elisa kit, indicated an 95% release of bFGF in 7 d and long-term sustained release of TGFß3. For alveolar defects rat models, the expression levels of CD29 and CD45, the bone volume fraction, trabecular number, and trabecular thickness of new bone monitored by Micro-CT in DFH treatment groups were significantly higher than others (*P < 0.05, vs Model). HE and Masson staining show the same results. In conclusion, DFH is a design of bionic alveolar remodelling microenvironment, that is in early time microvessels formed by bFGF provide nutritious to recruited endogenous stem cells, then TGFß3 slowly released speed up the process of new bones formation to common facilitate rat alveolar defect repair. The DFH with higher regenerative efficiency dovetails nicely with great demand due to the requirement of complicated biological processes.

Recombinant expression a novel fibronectin-collage fusion peptide modulating stem cell stemness via integrin ß3.

Constructing bionic extracellular matrix (ECM) is an attractive proposition for tissue engineering and clinical regeneration therapy involving the stemness of stem cells. Here, a novel recombinant protein fibronectin-collagen peptide (FCP) was designed to modulate the function of ECM expressed by Picha. pastoris strain X33. This FCP promotes cell migration and adhesion and maintains rBMSC stemness by binding integrin ß3. Its effects were blocked by both integrin ß3 siRNA and the integrin ß3 inhibitor Cilengitide. A template-independent ab initio prediction modeling approach is the best approach to construct a stable FCP protein model, which predicts the binding sites between FCP and integrin ß3. FCP may be used in the in vitro culture and clinical regeneration of stem cells that highly express integrin ß3, such as hematopoietic stem cells. The study provides information on the molecular structure of FCP and its bioactivity, which can be used to design new compounds. KEY POINTS: • Design a novel recombinant fibronectin-collagen peptide biomimetic ECM. • FCP promotes cell adhesion, migration, and proliferation. • Predicted and verified FCP structure and affinity with integrin ß3. • FCP binds integrin ß3 to maintain rBMSC stemness.