Additional file 1 of Aligned electrospun poly(l-lactide) nanofibers facilitate wound healing by inhibiting macrophage M1 polarization via the JAK-STAT and NF-κB pathways

Additional file 1: Table S1. Gene primer sequences for q-PCR. Fig. S1. Diameter distribution of electrospun fibers of A20 and R20 groups. Fig. S2. (A & B) Immunofluorescence staining of iNOS at days 7 and 14. (C & D) Mean fluorescence intensity of iNOS in each group of skin sections on days 7 and 14. Nucleus was stained blue and iNOS was stained red. Scale bar = 200 μm. Fig. S3. CCK-8 assay of MAEC culturing with macrophage conditioned medium. (*p < 0.05, n=3). Fig. S4. Migration of MAEC under conditional medium. Scale bar = 500 μm. Fig. S5. The expression of angiogenesis related genes of MAEC cultured with macrophage conditioned medium. (*p < 0.05, **p < 0.01, n = 3

Construction of a Hierarchical Micro-/Submicro-/Nanostructured 3D-Printed Ti6Al4V Surface Feature to Promote Osteogenesis: Involvement of Sema7A through the ITGB1/FAK/ERK Signaling Pathway

Constructing hierarchical hybrid structures is considered a facile method to improve the osseointegration of implants. Herein, a hierarchical micro-/submicro-/nanostructured surface feature of Ti6Al4V implants (3DAT group) was successfully constructed by combining the inherently formed three-dimensional (3D)-printed microscale topography, acid-etched sub-micropits, and anodized nanotubes. Compared with the classical SLA surface, the microscale topography and sub-micropits increased the three-dimensional space for the cell growth and mechanical stability of implants, while the modification of nanotubes dramatically improved the surface hydrophilicity, protein adsorption, and biomineralization. Most importantly, the 3DAT surface feature possessed excellent osteogenic performance in vitro and in vivo, with the involvement of semaphorin 7A (Sema7A) as revealed by RNA-seq through the ITGB1/FAK/ERK signaling pathway. The present study suggested that the hierarchically structured surface design strategy could accelerate the osseointegration rate of 3D-printed Ti6Al4V implants, promising personalized reconstruction of bone defects

DataSheet1_HIF-1α Regulates Osteogenesis of Periosteum-Derived Stem Cells Under Hypoxia Conditions via Modulating POSTN Expression.docx

Periosteum is indispensable in bone repair and is an important source of skeletal stem cells (SSCs) for endogenous bone regeneration. However, there are only a few studies about SSCs in periosteum. The craniomaxillofacial bone regeneration is done under the hypoxia microenvironment, in which HIF-1α plays an important role. The effect of HIF-1α on periosteum-derived stem cells (PDSCs) and the mechanisms of PDSCs activation under hypoxia conditions are unknown. In this study, the calvarial bone defect was established, with the periosteum removed or retained. Results show that the bone regeneration was severely impaired in the periosteum removed group. Moreover, pluripotent PDSCs isolated from the periosteum were positive for mesenchymal stem cell (MSC) markers. To determine the role of HIF-1α, the expression of HIF-1α was knocked down in vivo and in vitro, impairing the bone regeneration or osteogenesis of PDSCs. Furthermore, the knockdown of HIF-1α expression also reduced periostin (POSTN) expression, and recombinant POSTN addition partly rescued the osteogenic inhibition. Finally, to explore the mechanism under POSTN activation, the phosphorylation level of the PI3K/AKT pathway was assessed in transfected PDSCs. The phosphorylation level of PI3K and AKT was enhanced with HIF-1α overexpression and inhibited with HIF-1α knockdown, and the addition of PI3K activator or AKT activator could partly rescue POSTN expression. In conclusion, as a potential target to promote bone repair under the hypoxia microenvironment, HIF-1α can regulate the osteogenic differentiation of PDSCs via the PI3K/AKT/POSTN pathway, which lay a solid foundation for periosteum-based craniomaxillofacial bone regeneration.</p

The Effect of Quercetin on the Osteogenesic Differentiation and Angiogenic Factor Expression of Bone Marrow-Derived Mesenchymal Stem Cells

<div><p>Bone marrow-derived mesenchymal stem cells (BMSCs) are widely used in regenerative medicine in light of their ability to differentiate along the chondrogenic and osteogenic lineages. As a type of traditional Chinese medicine, quercetin has been preliminarily reported to promote osteogenic differentiation in osteoblasts. In the present study, the effects of quercetin on the proliferation, viability, cellular morphology, osteogenic differentiation and angiogenic factor secretion of rat BMSCs (rBMSCs) were examined by MTT assay, fluorescence activated cell sorter (FACS) analysis, real-time quantitative PCR (RT-PCR) analysis, alkaline phosphatase (ALP) activity and calcium deposition assays, and Enzyme-linked immunosorbent assay (ELISA). Moreover, whether mitogen-activated protein kinase (MAPK) signaling pathways were involved in these processes was also explored. The results showed that quercetin significantly enhanced the cell proliferation, osteogenic differentiation and angiogenic factor secretion of rBMSCs in a dose-dependent manner, with a concentration of 2 μM achieving the greatest stimulatory effect. Moreover, the activation of the extracellular signal-regulated protein kinases (ERK) and p38 pathways was observed in quercetin-treated rBMSCs. Furthermore, these induction effects could be repressed by either the ERK inhibitor PD98059 or the p38 inhibitor SB202190, respectively. These data indicated that quercetin could promote the proliferation, osteogenic differentiation and angiogenic factor secretion of rBMSCs in vitro, partially through the ERK and p38 signaling pathways.</p></div

Library Screening to Identify Highly-Effective Autophagy Inhibitors for Improving Photothermal Cancer Therapy

The small molecular inhibitor-associated downregulation of autophagy can remarkably enhance the efficiency of photothermal cancer therapy. To identify a more effective autophagy inhibitor, we screened a library of 20 compounds and found chloroquine, hydroxychloroquine, dauricine, and daurisoline were more efficient than the others to improve the photothermal killing of cancer cells. Interestingly, the four agents all disturb the autophagosome formation and fusion process, indicating it is a promising target to enhance cancer therapeutic efficiency. Among the four agents, daurisoline was identified to be the most efficient one. It reduced the viability of cancer cells treated by low-energy photothermal therapy from 86.27% to 32.92%. Finally, the combination treatment mediated by nanodrugs loaded with daurisoline and indocyanine green was more efficient than the individual modalities, resulting in complete inhibition of tumor growth. The study gives new inspiration to autophagy modulation-associated photothermal therapy and other therapeutic modalities for cancer treatment

Two-Dimensional Borocarbonitride Nanosheet-Engineered Hydrogel as an All-In-One Platform for Melanoma Therapy and Skin Regeneration

The postoperative tumor recurrence and repairing skin defects in clinical melanoma therapy remain challenging. Recent years have seen the development of visible-to-near-infrared (NIR) light for melanoma therapy or tissue regeneration. For solving the integrated issue of melanoma treatment and skin wounds repair, a gentle and efficient strategy is essential to utilize the multifunction of light. Here, we presented a new light-mediation concept and reported a light-responsive intelligent hydrogel system by introducing two-dimensional (2D) borocarbonitride (BCN) nanosheets into the methacrylated hyaluronic acid (HA) matrix (HA@BCN). The hydrogel was skillfully fabricated under the activation of blue light and exhibited excellent biocompatibility, mechanical robustness, and biodegradability, and then, a gentle and powerful multifunction for cutaneous melanoma therapy and wound healing under NIR light irradiation was performed. Based on this result, multifunctional hydrogels could be triggered by NIR light (0.35 W/cm2) for killing tumor cells, at least an 80% mortality rate in 10 min. Subsequently, the HA@BCN hydrogel could release more boron moieties as the growth promoter under moderate NIR light irradiation, which largely accelerated the wound healing. Therefore, our discovery presented a light-mediated and 2D nanomaterial-functionalized versatile hydrogel system for cutaneous melanoma photothermal therapy

The calcium deposition analysis.

<p>(A-B) ARS staining of rBMSCs cultured in DMEM medium (A) and osteogenic medium (B) with quercetin at different concentrations (0, 1, 2, 5 and 10 μM) for 28 days; (C-D) Calcium concentration analysis of rBMSCs cultured in DMEM medium (C) and osteogenic medium (D) with quercetin at different concentrations (0, 1, 2, 5 and 10 μM) for 21 and 28 days. *p < 0.05 indicates the quercetin-treated groups vs the control group; ▲p < 0.05 indicates the 2 μM group vs the other quercetin groups.</p

The ALP activity analysis.

<p>(A) ALP staining of rBMSCs cultured in DMEM with quercetin at different concentrations (0, 1, 2, 5 and 10 μM) for 10 days; (B) Quantitative analysis of ALP activity of rBMSCs cultured in DMEM with quercetin at different concentrations (0, 1, 2, 5 and 10 μM) for 4, 7 and 10 days. *p < 0.05 indicates quercetin-treated groups vs the control group; ^▲p < 0.05 indicates the 2 μM group vs the other quercetin groups.</p

Image1_Osteogenic differentiation of 3D-printed porous tantalum with nano-topographic modification for repairing craniofacial bone defects.TIF

Introduction: Congenital or acquired bone defects in the oral and cranio-maxillofacial (OCMF) regions can seriously affect the normal function and facial appearance of patients, and cause great harm to their physical and mental health. To achieve good bone defect repair results, the prosthesis requires good osteogenic ability, appropriate porosity, and precise three-dimensional shape. Tantalum (Ta) has better mechanical properties, osteogenic ability, and microstructure compared to Ti6Al4V, and has become a potential alternative material for bone repair. The bones in the OCMF region have unique shapes, and 3D printing technology is the preferred method for manufacturing personalized prosthesis with complex shapes and structures. The surface characteristics of materials, such as surface morphology, can affect the biological behavior of cells. Among them, nano-topographic surface modification can endow materials with unique surface properties such as wettability and large surface area, enhancing the adhesion of osteoblasts and thereby enhancing their osteogenic ability.Methods: This study used 3D-printed porous tantalum scaffolds, and constructed nano-topographic surface through hydrothermal treatment. Its osteogenic ability was verified through a series of in vitro and in vivo experiments.Results: The porous tantalum modified by nano-topographic surface can promote the proliferation and osteogenic differentiation of BMSCs, and accelerate the formation of new bone in the Angle of the mandible bone defect of rabbits.Discussion: It can be seen that 3D-printed nano-topographic surface modified porous tantalum has broad application prospects in the repair of OCMF bone defects.</p