Nanohydroxyapatite Stimulates PD-L1 Expression to Boost Melanoma Combination Immunotherapy.

Although checkpoint-inhibitor immunotherapy held tremendous advances, improving immune response during treatment has always been an urgent clinical issue. With the help of mRNA microarray technology, it was found that short rod-like nanohydroxyapatite (nHA) promoted the upregulation of CD274 and PD-L1 related gene transcription, which was confirmed by the significantly enhanced PD-L1 expression level in B16, B16F10, and 4T1 cells in vitro. Hence, an injectable in situ responsive hydrogel reservoir embed with nHA and PD-1/PD-L1 inhibitor was engineered for a combination immunotherapy by peritumoral administration. The results confirmed that the combinational strategy effectively suppressed tumorigenesis and tumor growth, recovered the abnormal lactate dehydrogenase, aspartate transaminase, and alanine aminotransferase indicators, and significantly elongated the life span of a tumor-bearing mouse. The substantive progress mainly derived from nHA-induced T cell infiltration reinforcement in a tumor site and CD8+ T cell polarization in spleen, implying that nHA might function as an immunomodulator for melanoma immunotherapy.

Dynamic mechanical loading facilitated chondrogenic differentiation of rabbit BMSCs in collagen scaffolds.

Mechanical signals have been played close attention to regulate chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). In this study, dynamic mechanical loading simulation with natural frequencies and intensities were applied to the 3D cultured BMSCs-collagen scaffold constructs. We investigated the effects of dynamic mechanical loading on cell adhesion, uniform distribution, proliferation, secretion of extracellular matrix (ECM) and chondrogenic differentiation of BMSCs-collagen scaffold constructs. The results indicated that dynamic mechanical loading facilitated the BMSCs adhesion, uniform distribution, proliferation and secretion of ECM with a slight contraction, which significantly improved the mechanical strength of the BMSCs-collagen scaffold constructs for better mimicking the structure and function of a native cartilage. Gene expression results indicated that dynamic mechanical loading contributed to the chondrogenic differentiation of BMSCs with higher levels of AGG, COL2A1 and SOX9 genes, and prevented of hypertrophic process with lower levels of COL10A1, and reduced the possibility of fibrocartilage formation due to down-regulated COL1A2. In conclusion, this study emphasized the important role of dynamic mechanical loading on promoting BMSCs chondrogenic differentiation and maintaining the cartilage phenotype for in vitro reconstruction of tissue-engineered cartilage, which provided an attractive prospect and a feasibility strategy for cartilage repair.

Temperature and ion dual responsive biphenyl-dipeptide supramolecular hydrogels as extracellular matrix mimic-scaffolds for cell culture applications.

Stimuli-responsive supramolecular hydrogels composed of aromatic short peptide gelators have attracted intensive attention in the field of biomedicine because of their stable chemical structure, simple and convenient synthetic route and intelligent response to external stimuli. In this paper, several dipeptides were coupled to biphenylacetic acid (BPAA) to generate aromatic short peptide compounds through the standard solid phase peptide synthesis. These BPAA-dipeptide compounds presented clearly different gelation behaviors from the generally employed Fmoc-dipeptide and Nap-dipeptide compounds, but only BPAA-diphenylalanine was able to form homogeneous and transparent hydrogels through temperature switching or ion induction. Utilizing the biphenyl group not only expanded the scope of aromatic molecules serving as building blocks of aromatic short peptide gelators but also demonstrated the critical role of aromatic molecules in the self-assembling process. Moreover, supramolecular hydrogels initiated by heating-cooling or salt addition could be exploited as extracellular matrix (ECM) mimic scaffolds to support the adhesive growth and proliferation of L929 cells in 2D/3D culture under physiological conditions, demonstrating their potential applications in regenerative medicine.

Extracellular matrix powder from cultured cartilage-like tissue as cell carrier for cartilage repair.

Cartilage extracellular matrix (ECM) is a promising material for cartilage repair because of its bioactivity. However, the animal source of ECM unavoidably increases the risk of pathogen infection and the variability of product quality. In this study, we utilized a novel 3D culture method to prepare a new type of artificial decellularized matrix powder (DEMP) for the development of injectable, bioactive, biodegradable cell carriers for cartilage tissue engineering. This culture method combined hanging drop culture with suspension culture method, and was very efficient to produce cartilage-like tissue (CLT). By this method, an initial 2.3 × 106 chondrocyte generated as much as 58.22 mg wet weight CLT at two weeks, which proved to contain abundant glycoaminoglycans (GAGs), type II collagen, and BMP-2 and TGF-ß1 growth factors by staining techniques and biochemical analysis. Subsequently, the two-week-old CLT was decellularized to prepare the artificial DEMP. In an in vitro study, it was found that MSCs cultured on DEMP differentiated to chondrocytes very well and secreted rich GAGs and type II collagen at three weeks even without exogenous TGF-ß1. The in vivo study demonstrated that the DEMP not only facilitated regeneration of hyaline cartilage, which was implied by the intense staining of GAGs and type II collagen in rabbit subchondral defects at 1 month, but also benefited the regeneration of subchondral bone (bone ingrowth at 1 month: 48.22%) as shown in micro-CT data. Collectively, these results suggest that the artificial DEMP prepared by this culture method holds great potential as a novel ECM material for cartilage repair.

Tough and elastic hydrogel of hyaluronic acid and chondroitin sulfate as potential cell scaffold materials.

Natural polysaccharides are extensively investigated as cell scaffold materials for cellular adhesion, proliferation, and differentiation due to their excellent biocompatibility, biodegradability, and biofunctions. However, their application is often severely limited by their mechanical behavior. In this study, a tough and elastic hydrogel scaffold was prepared with hyaluronic acid (HA) and chondroitin sulfate (CS). HA and CS were conjugated with tyramine (TA) and the degree of substitution (DS) was 10.7% and 11.3%, respectively, as calculated by (1)H NMR spectra. The hydrogel was prepared by mixing HA-TA and CS-TA in presence of H2O2 and HRP. The sectional morphology of hydrogels was observed by SEM, static and dynamic mechanical properties were analyzed by Shimadzu electromechanical testing machine and dynamic mechanical thermal analyzer Q800. All samples showed good ability to recover their appearances after deformation, the storage modulus (E') of hydrogels became higher as the testing frequency went up. Hydrogels also showed fatigue resistance to cyclic compression. Mesenchymal stem cells encapsulated in hydrogels showed good cell viability as detected by CLSM. This study suggests that the hydrogels have both good mechanical properties and biocompatibility, and may serve as model systems to explore mechanisms of deformation and energy dissipation or find some applications in tissue engineering.