IL-7-primed bystander CD8 tumor-infiltrating lymphocytes optimize the antitumor efficacy of T cell engager immunotherapy.

Bispecific T cell engagers (TCEs) show promising clinical efficacy in blood tumors, but their application to solid tumors remains challenging. Here, we show that Fc-fused IL-7 (rhIL-7-hyFc) changes the intratumoral CD8 T cell landscape, enhancing the efficacy of TCE immunotherapy. rhIL-7-hyFc induces a dramatic increase in CD8 tumor-infiltrating lymphocytes (TILs) in various solid tumors, but the majority of these cells are PD-1-negative tumor non-responsive bystander T cells. However, they are non-exhausted and central memory-phenotype CD8 T cells with high T cell receptor (TCR)-recall capacity that can be triggered by tumor antigen-specific TCEs to acquire tumoricidal activity. Single-cell transcriptome analysis reveals that rhIL-7-hyFc-induced bystander CD8 TILs transform into cycling transitional T cells by TCE redirection with decreased memory markers and increased cytotoxic molecules. Notably, TCE treatment has no major effect on tumor-reactive CD8 TILs. Our results suggest that rhIL-7-hyFc treatment promotes the antitumor efficacy of TCE immunotherapy by increasing TCE-sensitive bystander CD8 TILs in solid tumors.

Metabolomics and miRNA profiling reveals feature of gallbladder cancer-derived biliary extracellular vesicles.

BACKGROUND: Although there are several studies in the development of various human cancers, the role of exosomes is poorly understood in the progression of gallbladder cancer. This study aims to characterize the metabolic changes occurring in exosomes obtained from patients with gallbladder cancer compared with those from other gallbladder disease groups. METHODS: Biliary exosomes were isolated from healthy donors (n = 3) and from patients with gallbladder cancer (n = 3), gallbladder polyps (n = 4), or cholecystitis (n = 3) using a validated exosome isolation kit. Afterward, we performed miRNA profiling and untargeted metabolomic analysis of the exosomes. The results were validated by integrating the results of the miRNA and metabolomic analyses. RESULTS: The gallbladder cancer group exhibited a significant reduction in the levels of multiple unsaturated phosphatidylethanolamines and phosphatidylcholines compared to the normal group, which resulted in the loss of exosome membrane integrity. Additionally, the gallbladder cancer group demonstrated significant overexpression of miR-181c and palmitic acid, and decreased levels of conjugated deoxycholic acid, all of which are strongly associated with the activation of the PI3K/AKT pathway. CONCLUSIONS: Our findings demonstrate that the contents of exosomes are disease-specific, particularly in gallbladder cancer, and that altered metabolites convey critical information regarding their phenotype. We believe that our metabolomic and miRNA profiling results may provide important insights into the development of gallbladder cancer.

Characteristics of Hf0.5Zr0.5O2 Thin Films Prepared by Direct and Remote Plasma Atomic Layer Deposition for Application to Ferroelectric Memory.

Hf0.5Zr0.5O2 (HZO) thin film exhibits ferroelectric properties and is presumed to be suitable for use in next-generation memory devices because of its compatibility with the complementary metal-oxide-semiconductor (CMOS) process. This study examined the physical and electrical properties of HZO thin films deposited by two plasma-enhanced atomic layer deposition (PEALD) methods- direct plasma atomic layer deposition (DPALD) and remote plasma atomic layer deposition (RPALD)-and the effects of plasma application on the properties of HZO thin films. The initial conditions for HZO thin film deposition, depending on the RPALD deposition temperature, were established based on previous research on HZO thin films deposited by the DPALD method. The results show that as the measurement temperature increases, the electric properties of DPALD HZO quickly deteriorate; however, the RPALD HZO thin film exhibited excellent fatigue endurance at a measurement temperature of 60 °C or less. HZO thin films deposited by the DPALD and RPALD methods exhibited relatively good remanent polarization and fatigue endurance, respectively. These results confirm the applicability of the HZO thin films deposited by the RPALD method as ferroelectric memory devices.

Effect of Process Temperature on Density and Electrical Characteristics of Hf0.5Zr0.5O2 Thin Films Prepared by Plasma-Enhanced Atomic Layer Deposition.

HfxZr1-xO2 (HZO) thin films have excellent potential for application in various devices, including ferroelectric transistors and semiconductor memories. However, such applications are hindered by the low remanent polarization (Pr) and fatigue endurance of these films. To overcome these limitations, in this study, HZO thin films were fabricated via plasma-enhanced atomic layer deposition (PEALD), and the effects of the deposition and post-annealing temperatures on the density, crystallinity, and electrical properties of the thin films were analyzed. The thin films obtained via PEALD were characterized using cross-sectional transmission electron microscopy images and energy-dispersive spectroscopy analysis. An HZO thin film deposited at 180 °C exhibited the highest o-phase proportion as well as the highest density. By contrast, mixed secondary phases were observed in a thin film deposited at 280 °C. Furthermore, a post-annealing temperature of 600 °C yielded the highest thin film density, and the highest 2Pr value and fatigue endurance were obtained for the film deposited at 180 °C and post-annealed at 600 °C. In addition, we developed three different methods to further enhance the density of the films. Consequently, an enhanced maximum density and exceptional fatigue endurance of 2.5 × 107 cycles were obtained.

Adenine base editing and prime editing of chemically derived hepatic progenitors rescue genetic liver disease.

DNA base editors and prime editing technology enable therapeutic in situ correction of disease-causing alleles. These techniques could have broad applications for ex vivo editing of cells prior to transplantation in a range of diseases, but it is critical that the target population is efficiently modified and engrafts into the host. Chemically derived hepatic progenitors (CdHs) are a multipotent population capable of robust engraftment and hepatocyte differentiation. Here we reprogrammed hepatocytes from a mouse model of hereditary tyrosinemia type 1 (HT1) into expandable CdHs and successfully corrected the disease-causing mutation using both adenine base editors (ABEs) and prime editors (PEs). ABE- and PE-corrected CdHs repopulated the liver with fumarylacetoacetate hydrolase-positive cells and dramatically increased survival of mutant HT1 mice. These results demonstrate the feasibility of precise gene editing in transplantable cell populations for potential treatment of genetic liver disease.

Anterior cruciate ligament remnant-preserving and re-tensioning reconstruction: a biomechanical comparison study of three different re-tensioning methods in a porcine model.

BACKGROUND: With the developments in the arthroscopic technique, anterior cruciate ligament (ACL) remnant-preserving reconstruction is gradually gaining attention with respect to improving proprioception and enhancing early revascularization of the graft. To evaluate the mechanical pull-out strength of three different methods for remnant-preserving and re-tensioning reconstruction during ACL reconstruction. METHODS: Twenty-seven fresh knees from mature pigs were used in this study. Each knee was dissected to isolate the femoral attachment of ACL and cut the attachment. An MTS tensile testing machine with dual-screw fixation clamp with 30° flexion angle was used. The 27 specimens were tested after applying re-tensioning sutures with No. 0 polydioxanone (PDS), using the single stitch (n = 9), loop stitch (n = 9), and triple stitch (n = 9) methods. We measured the mode of failure, defined as (1) ligament failure (longitudinal splitting of the remnant ACL) or (2) suture failure (tearing of the PDS stitch); load-to-failure strength; and stiffness for the three methods. Kruskal-Wallis test and Mann-Whitney U-test were used to compare the variance of load-to-failure strength and stiffness among the three groups. RESULTS: Ligament failure occurred in all cases in the single stitch group and in all but one case in the triple stitch group. Suture failure occurred in all cases in the loop stitch group and in one case in the triple stitch group. The load-to-failure strength was significantly higher with loop stich (91.52 ± 8.19 N) and triple stitch (111.1 ± 18.15 N) than with single stitch (43.79 ± 11.54 N) (p = 0.002). With respect to stiffness, triple stitch (2.50 ± 0.37 N/mm) yielded significantly higher stiffness than the other methods (p = 0.001). CONCLUSIONS: The results suggested that loop stitch or triple stitch would be a better option for increasing the mechanical strength when applying remnant-preserving and re-tensioning reconstruction during ACL reconstruction.

Efficacy of bone formation of microporous sphere-shaped biphasic calcium phosphate in a rabbit skull bone defect model.

Bone graft is required in various surgical procedures. Although autograft is the gold standard, it has limited availability. Various compounds have been proposed as alternatives such as biphasic calcium phosphate (BCP), which is the most widely used compound. The newly synthesized microporous sphere-shaped BCP has the advantage of increasing contact surface, and it can induce the formation of microbone structures. A putty-type contains the addition of a fluid carrier to the sphere-shaped BCP and can be easily used for a small orifice large bone defect. To compare the widely used BCP products, new bone formation and residual graft materials (RGM) were evaluated for 6 and 12 weeks in a rabbit calvarial bone defect model. Although existing BCP products and the microporous sphere-type product did not differ significantly with respect to new bone formation and RGM, the putty-type product was largely washed out and had low new bone formation at 6 and 12 weeks. Overall, the results suggest that microporous sphere-shaped BCP showed similar bone formation capability to existing products and was able to maintain higher initial mechanical stability.

3D cell-printing of biocompatible and functional meniscus constructs using meniscus-derived bioink.

Meniscus injuries are prevalent in orthopedic diagnosis. The reconstruction of the structural inhomogeneity and anisotropy of the meniscus is a major challenge in clinical practice. Meniscal tissue engineering has emerged as a potential alternative for the treatment of various meniscal diseases and injuries. In this study, we developed three-dimensional (3D) cell-printed meniscus constructs using a mixture of polyurethane and polycaprolactone polymers and cell-laden decellularized meniscal extracellular matrix (me-dECM) bioink with high controllability and durable architectural integrity. The me-dECM bioink provided 3D cell-printed meniscus constructs with a conducive biochemical environment that supported growth and promoted the proliferation and differentiation of encapsulated stem cells toward fibrochondrogenic commitment. In addition, we investigated the in vivo performance of the 3D cell-printed meniscus constructs, which exhibited biocompatibility, excellent mechanical properties, and improved biological functionality. These attributes were similar to those of the native meniscus. Collectively, the 3D cell-printing technology and me-dECM bioink facilitate the recapitulation of meniscus tissue specificity in the aspect of the shape and microenvironment for meniscus regeneration. Further, the developed constructs can potentially be applied in clinical practice.

Osteo-Compatibility of 3D Titanium Porous Coating Applied by Direct Energy Deposition (DED) for a Cementless Total Knee Arthroplasty Implant: in Vitro and in Vivo Study.

: Direct energy deposition (DED) technology has gained increasing attention as a new implant surface technology that replicates the porous structure of natural bones facilitating osteoblast colonization and bone ingrowth. However, concerns have arisen over osteolysis or chronic inflammation that could be caused by Cobalt-chrome (CoCr) alloy and Titanium (Ti) nanoparticles produced during the fabrication process. Here, we evaluated whether a DED Ti-coated on CoCr alloy could improve osteoblast colonization and osseointegration in vitro and in vivo without causing any significant side effects. Three types of implant CoCr surfaces (smooth, sand-blasted and DED Ti-coated) were tested and compared. Three cell proliferation markers and six inflammatory cytokine markers were measured using SaOS2 osteoblast cells. Subsequently, X-ray and bone histomorphometric analyses were performed after implantation into rabbit femur. There were no differences between the DED group and positive control in cytokine assays. However, in the 5-bromo-2'-deoxyuridine (BrdU) assay the DED group exhibited even higher values than the positive control. For bone histomorphometry, DED was significantly superior within the 1000 µm bone area. The results suggest that DED Ti-coated metal printing does not affect the osteoblast viability or impair osseointegration in vitro and in vivo. Thus, this technology is biocompatible for coating the surfaces of cementless total knee arthroplasty (TKA) implants.

Titanium Porous Coating Using 3D Direct Energy Deposition (DED) Printing for Cementless TKA Implants: Does It Induce Chronic Inflammation?

Because of the recent technological advances, the cementless total knee arthroplasty (TKA) implant showed satisfactory implant survival rate. Newly developed 3D printing direct energy deposition (DED) has superior resistance to abrasion as compared to traditional methods. However, there is still concern about the mechanical stability and the risk of osteolysis by the titanium (Ti) nanoparticles. Therefore, in this work, we investigated whether DED Ti-coated cobalt-chrome (CoCr) alloys induce chronic inflammation reactions through in vitro and in vivo models. We studied three types of implant surfaces (smooth, sand-blasted, and DED Ti-coated) to compare their inflammatory reaction. We conducted the in vitro effect of specimens using the cell counting kit-8 (CCK-8) assay and an inflammatory cytokine assay. Subsequently, in vivo analysis of the immune profiling, cytokine assay, and histomorphometric evaluation using C57BL/6 mice were performed. There were no significant differences in the CCK-8 assay, the cytokine assay, and the immune profiling assay. Moreover, there were no difference for semi-quantitative histomorphometry analysis at 4 and 8 weeks among the sham, smooth, and DED Ti-coated samples. These results suggest that DED Ti-coated printing technique do not induce chronic inflammation both in vitro and in vivo. It has biocompatibility for being used as a surface coating of TKA implant.