Synergistic Integration of mRNA-LNP with CAR-Engineered Immune Cells: Pioneering Progress in Immunotherapy.

Chimeric antigen receptor T-cell (CAR-T) therapy has emerged as a revolutionary approach in the treatment of malignancies. Despite its remarkable successes, this field continues to grapple with challenges such as scalability, safety concerns, limited therapeutic effect, in vivo persistence, and the need for precise control over CAR expression. In the post-pandemic era of COVID-19 vaccine immunization, the application of messenger RNA (mRNA) encapsulated within lipid nanoparticles (LNP) has recently garnered significant attention as a potential solution to address these challenges. This review delves into the dynamic landscape of mRNA-LNP technology and its potential implications for CAR-engineered immune cells-based immunotherapy.

Patient-reported outcome measures and health economics in regenerative periodontal therapy: A systematic review and meta-analysis.

Understanding patient responses to periodontal regeneration is crucial. This systematic review and meta-analysis addressed two key questions: (a) the impact of periodontal regeneration on patient-reported outcome measures (PROMs) for intrabony and furcation involvement and (b) the cost-effectiveness of periodontal regeneration for treating periodontal defects. Twenty-four studies were included, with 20 randomized clinical trials (RCTs) reporting patient-reported outcomes and five (three RCTs and two economic model-based studies) reporting cost-effectiveness outcomes. Results favored regeneration therapy over conventional flap surgery for intrabony defects, showing improvements in qualitative (i.e., amount of regenerated attachment apparatus) and quantitative parameters (i.e., probing and radiographic parameters). In terms of PROMs, regenerative treatments involving barrier membranes resulted in longer chair times and higher rates of complications (such as membrane exposure or edema) compared to flap with biologic agents or access flap alone. Despite this, oral health-related quality of life improved after both regenerative and extraction procedures. Economically, regeneration remained favorable compared to extraction and replacement or open flap debridement alone for periodontal defects. Single-flap variants in open flap debridement yielded similar outcomes to regenerative treatment, offering a potentially cost-effective option. Nevertheless, further discussion on the benefits of less-invasive flap designs is needed due to the lack of histological evaluation.

A kinase fusion protein from Aegilops longissima confers resistance to wheat powdery mildew.

Many disease resistance genes have been introgressed into wheat from its wild relatives. However, reduced recombination within the introgressed segments hinders the cloning of the introgressed genes. Here, we have cloned the powdery mildew resistance gene Pm13, which is introgressed into wheat from Aegilops longissima, using a method that combines physical mapping with radiation-induced chromosomal aberrations and transcriptome sequencing analysis of ethyl methanesulfonate (EMS)-induced loss-of-function mutants. Pm13 encodes a kinase fusion protein, designated MLKL-K, with an N-terminal domain of mixed lineage kinase domain-like protein (MLKL_NTD domain) and a C-terminal serine/threonine kinase domain bridged by a brace. The resistance function of Pm13 is validated through transient and stable transgenic complementation assays. Transient over-expression analyses in Nicotiana benthamiana leaves and wheat protoplasts reveal that the fragment Brace-Kinase122-476 of MLKL-K is capable of inducing cell death, which is dependent on a functional kinase domain and the three α-helices in the brace region close to the N-terminus of the kinase domain.

Harmonizing the symphony of chimeric antigen receptor T cell immunotherapy with the elegance of biomaterials.

Chimeric antigen receptor T cell (CAR-T) immunotherapy has become a heated field of cancer research, demonstrating revolutionary efficacy in refractory and relapsed hematologic malignancies. However, CAR-T therapy has still encountered tough challenges, including complicated and lengthy manufacturing procedures, mediocre targeted delivery, limited therapeutic effect against solid tumors and difficulties in real-time in vivo monitoring. To overcome these limitations, various versatile biomaterials have been used in the above aspects and have improved CAR-T therapy impressively. This review mainly summarizes the latest research progress of biomaterials promoting CAR-T therapy in manufacturing, enhancing targeted delivery and tumor infiltration, and dramatic in vivo tracking to provide new insights and inspiration for clinical treatment.

Decision Tree for Reconstructive Treatment of Peri-implantitis Defects.

Peri-implantitis, a common complication among patients receiving implant-supported restorative therapy, often requires surgical intervention for effective treatment. Understanding the specific configuration of peri-implant bony defect and adjacent bone peaks is crucial for tailoring treatment strategies and improving outcomes. A decision tree for reconstructive peri-implantitis therapy has been developed based on the new classification of defect configurations (Class I to V), guiding clinicians in selecting treatment options, including biomaterials, techniques, and healing approaches. Furthermore, clinicians are encouraged to consider various factors such as local predisposing factors (such as soft tissue characteristics, prosthetic design, and implant position in three-dimensional perspective), clinical factors (surgeon skill and experience), and patient-related factors (such as local and systemic health, preferences, and cost) when evaluating reconstructive therapy options.

Calibrated intraoral scan protocol (CISP) for full-arch implant impressions: An in vitro comparison to conventional impression, intraoral scan, and intraoral scan with scan-aid.

OBJECTIVE: To assess a newly developed intraoral scan protocol in enhancing the accuracy of complete-arch implant impressions. MATERIALS AND METHODS: Four impression approaches were applied to the same maxillary edentulous model with 6 implants: (1) intraoral scan (IOS), (2) intraoral scan with scan aid (IOS-SA), (3) calibrated intraoral scan protocol (CISP), and (4) conventional splinted open-tray impression (CONV). Each approach was repeated 10 times, and a direct scan of the model with a desktop scanner was used as a reference model. The alignment of scans and the reference model was conducted by two methods: (a) aligning all scan bodies to evaluate the overall fit, and (b) aligning the first and second scan bodies to simulate the Sheffield fit test for passive fitting of multiple implant-supported prostheses. Linear deviations from the reference model (trueness) and within each group (precision) were analyzed using Python scripts. RESULTS: When aligned by all scan bodies, the CISP group exhibited comparable mean trueness (38.33 µm) and precision (45.97 µm) to the CONV group (44.30 and 47.92 µm respectively), both of which significantly outperformed the IOS group (86.82 and 83.17 µm, respectively). Furthermore, in the virtual Sheffield fit test, the CISP group achieved the highest levels of mean trueness at the end span (121.7 µm), making a linear deviation reduction of 36.7%, 60%, and 41.4% when compared to the CONV, the IOS, and the IOS-SA groups, respectively. Moreover, the CISP group (104.3 µm) displayed a remarkable 65, 182, and 86 µm advantage in precision over the CONV, IOS, and IOS-SA groups, respectively. CONCLUSION: CISP demonstrated comparable accuracy to the gold standard, the conventional splinted open-tray impression. Furthermore, it excelled in the virtual passive fitting test.

Accuracy of open-sleeved vs. closed-sleeved static computer-assisted implant systems in immediate maxillary molar implant placement: An in vitro study.

OBJECTIVES: The objective of this study is (1) to compare the accuracy of an open-sleeved static computer-assisted implant system (sCAIS) with a closed-sleeve sCAIS and free-hand approach in immediate implant placement (IIP) of maxillary molar sites and (2) to investigate the influence of socket morphology on these approaches. MATERIALS AND METHODS: Ninety partially edentulous duplicated maxillary models simulating three different molar sockets (type A, B, and C based on Smith and Tarnow's classification) were investigated. Three modalities, including sCAIS with open-sleeves, sCAIS with closed-sleeves, and free-hand approach, were applied separately to 30 models with 120 sockets. A customized Python script automatically measured the deviations between the virtual and actual implant positions for all 360 implants. RESULTS: The 3D deviations of sCAIS were significantly influenced by the socket and sleeve types. Both guided groups exhibited significantly less deviation than the free-hand approach. Type A and C sockets resulted in better implant positions than type B socket sites. In type B sockets, the open-sleeve group achieved significantly less deviation compared to the closed-sleeve group, with respect to apical global (1.34 ± 0.53 vs. 1.84 ± 0.59 mm), coronal horizontal (0.68 ± 0.36 vs. 0.93 ± 0.34 mm), apical horizontal (1.21 ± 0.59 vs. 1.74 ± 0.63 mm), and angular (3.30 ± 1.41 vs. 4.41 ± 1.96°) deviations. CONCLUSIONS: Guided implant surgery significantly reduces deviations during molar IIP compared to free-hand procedures. Furthermore, the use of open-sleeve sCAIS appears to be more effective in minimizing deviations in type B sockets when compared with the closed-sleeve guided system.

Co-delivery of fucoxanthin and Twist siRNA using hydroxyethyl starch-cholesterol self-assembled polymer nanoparticles for triple-negative breast cancer synergistic therapy.

INTRODUCTION: Triple-negative breast cancer (TNBC) represents the most aggressive subtype of breast cancer with an extremely dismal prognosis and few treatment options. As a desmoplastic tumor, TNBC tumor cells are girdled by stroma composed of cancer-associated fibroblasts (CAFs) and their secreted stromal components. The rapidly proliferating tumor cells, together with the tumor stroma, exert additional solid tissue pressure on tumor vasculature and surrounding tissues, severely obstructing therapeutic agent from deep intratumoral penetration, and resulting in tumor metastasis and treatment resistance. OBJECTIVES: Fucoxanthin (FX), a xanthophyll carotenoid abundant in marine algae, has attracted widespread attention as a promising alternative candidate for tumor prevention and treatment. Twist is a pivotal regulator of epithelial to mesenchymal transition, and its depletion has proven to sensitize antitumor drugs, inhibit metastasis, reduce CAFs activation and the following interstitial deposition, and increase tumor perfusion. The nanodrug delivery system co-encapsulating FX and nucleic acid drug Twist siRNA (siTwist) was expected to form a potent anti-TNBC therapeutic cyclical feedback loop. METHODS AND RESULTS: Herein, our studies constituted a novel self-assembled polymer nanomedicine (siTwist/FX@HES-CH) based on the amino-modified hydroxyethyl starch (HES-NH2) grafted with hydrophobic segment cholesterol (CH). The MTT assay, flow cytometry apoptosis analysis, transwell assay, western blot, and 3D multicellular tumor spheroids growth inhibition assay all showed that siTwist/FX@HES-CH could kill tumor cells and inhibit their metastasis in a synergistic manner. The in vivo anti-TNBC efficacy was demonstrated that siTwist/FX@HES-CH remodeled tumor microenvironment, facilitated interstitial barrier crossing, killed tumor cells synergistically, drastically reduced TNBC orthotopic tumor burden and inhibited lung metastasis. CONCLUSION: Systematic studies revealed that this dual-functional nanomedicine that targets both tumor cells and tumor microenvironment significantly alleviates TNBC orthotopic tumor burden and inhibits lung metastasis, establishing a new paradigm for TNBC therapy.

The enchanting canvas of CAR technology: Unveiling its wonders in non-neoplastic diseases.

Chimeric antigen receptor (CAR) T cells have made a groundbreaking advancement in personalized immunotherapy and achieved widespread success in hematological malignancies. As CAR technology continues to evolve, numerous studies have unveiled its potential far beyond the realm of oncology. This review focuses on the current applications of CAR-based cellular platforms in non-neoplastic indications, such as autoimmune, infectious, fibrotic, and cellular senescence-associated diseases. Furthermore, we delve into the utilization of CARs in non-T cell populations such as natural killer (NK) cells and macrophages, highlighting their therapeutic potential in non-neoplastic conditions and offering the potential for targeted, personalized therapies to improve patient outcomes and enhanced quality of life.

A skeleton in a huff: insights in etiologies of osteosclerosis.

A 30-yr-old man developed right lower leg pain and a palpable solid mass. Radiographic imaging revealed a periosteal reaction with an exostotic mass arising from the right distal fibula. Generalized skeletal osteosclerosis with periosteal reaction was discovered on a radiographic skeletal survey. A biopsy of the right fibular mass revealed reactive woven bone. The patient was referred to a metabolic bone disease clinic, where laboratory values were consistent with secondary hyperparathyroidism and increased bone turnover. A DXA bone density scan revealed high bone density, with an L1-4 spine Z-score of +9.3, a left femoral neck Z-score of +8.5, and a total hip Z-score of +6.5. A dental exam revealed generalized gingival inflammation, teeth mobility, generalized horizontal alveolar bone loss and widening of the periodontal ligament space, increased bone density around the teeth, and thickening of the radicular lamina dura. An extensive evaluation was performed, with the result of a single test revealing the diagnosis. The differential diagnoses of osteosclerosis affecting the skeleton, teeth, and oral cavity are discussed.

A 30-yr-old man developed, over a short period, pain in his lower right leg accompanied by a hard mass. He also reported weight loss and night sweats for the past 6 months. After evaluation by his primary physician, an X-ray was ordered that reported a bony mass arising from the right fibula bone. A biopsy was performed of the mass, but no evidence of cancer or any other specific abnormality was found. The patient was then referred to a bone disease specialty clinic. Laboratory tests revealed a large increase in how quickly the patient's skeleton was remodeling, affecting the balance of bone formation and removal involved in maintaining a healthy skeleton. A bone density scan reported that the patient had very dense bones. Other unusual changes were also discovered in a dental exam, suggesting bone thickening. After an extensive evaluation, a single blood test revealed the cause of the fibular bone mass and dense bones.

Targeting autophagy overcomes cancer-intrinsic resistance to CAR-T immunotherapy in B-cell malignancies.

BACKGROUND: Chimeric antigen receptor T (CAR-T) therapy has substantially revolutionized the clinical outcomes of patients with hematologic malignancies, but the cancer-intrinsic mechanisms underlying resistance to CAR-T cells remain yet to be fully understood. This study aims to explore the molecular determinants of cancer cell sensitivity to CAR-T cell-mediated killing and to provide a better understanding of the underlying mechanisms and potential modulation to improve clinical efficacy. METHODS: The human whole-genome CRISPR/Cas9-based knockout screening was conducted to identify key genes that enable cancer cells to evade CD19 CAR-T-cell-mediated killing. The in vitro cytotoxicity assays and evaluation of tumor tissue and bone marrow specimens were further conducted to confirm the role of the key genes in cancer cell susceptibility to CAR-T cells. In addition, the specific mechanisms influencing CAR-T cell-mediated cancer clearance were elucidated in mouse and cellular models. RESULTS: The CRISPR/Cas9-based knockout screening showed that the enrichment of autophagy-related genes (ATG3, BECN1, and RB1CC1) provided protection of cancer cells from CD19 CAR-T cell-mediated cytotoxicity. These findings were further validated by in vitro cytotoxicity assays in cells with genetic and pharmacological inhibition of autophagy. Notably, higher expression of the three autophagy-related proteins in tumor samples was correlated with poorer responsiveness and worse survival in patients with relapsed/refractory B-cell lymphoma after CD19 CAR-T therapy. Bulk RNA sequencing analysis of bone marrow samples from B-cell leukemia patients also suggested the clinical relevance of autophagy to the therapeutic response and relapse after CD19 CAR-T cell therapy. Pharmacological inhibition of autophagy and knockout of RB1CC1 could dramatically sensitize tumor cells to CD19 CAR-T cell-mediated killing in mouse models of both B-cell leukemia and lymphoma. Moreover, our study revealed that cancer-intrinsic autophagy mediates evasion of CAR-T cells via the TNF-α-TNFR1 axis-mediated apoptosis and STAT1/IRF1-induced chemokine signaling activation. CONCLUSIONS: These findings confirm that autophagy signaling in B-cell malignancies is essential for the effective cytotoxic function of CAR-T cells and thereby pave the way for the development of autophagy-targeting strategies to improve the clinical efficacy of CAR-T cell immunotherapy.

Harnessing Biomaterials for Safeguarding Chimeric Antigen Receptor T Cell Therapy: An Artful Expedition in Mitigating Adverse Effects.

Chimeric antigen receptor T cell (CAR-T) therapy has emerged as a groundbreaking approach in cancer treatment, showcasing remarkable efficacy. However, the formidable challenge lies in taming the formidable side effects associated with this innovative therapy, among which cytokine release syndrome (CRS), immune effector cell-associated neurotoxicity syndrome (ICANS) and on-target off-tumor toxicities (OTOT) are typical representatives. Championing the next frontier in cellular immunotherapy, this comprehensive review embarks on an artistic exploration of leveraging biomaterials to meticulously navigate the intricate landscape of CAR-T cell therapy. Unraveling the tapestry of potential toxicities, our discourse unveils a symphony of innovative strategies designed to elevate the safety profile of this revolutionary therapeutic approach. Through the lens of advanced medical science, we illuminate the promise of biomaterial interventions in sculpting a safer and more efficacious path for CAR-T cell therapy, transcending the boundaries of conventional treatment paradigms.

Recommendations for successful virtual patient-assisted esthetic implant rehabilitation: A guide for optimal function and clinical efficiency.

OBJECTIVE: Complete arch implant rehabilitation necessitates meticulous treatment planning and high-level collaboration between surgical and prosthetic dental teams. Emerging virtual technologies hold considerable promise in streamlining this process. The aim of this article is to extend recommendations to clinicians venturing into the virtual patient-assisted esthetic implant rehabilitation workflow. OVERVIEW: This article summarizes recommendations for virtual patient-assisted esthetic implant rehabilitation in the following five aspects: three-dimensional data handling and superimposition, occlusion and virtual articulator integration in creating virtual patients, streamlined face- and prosthetic-driven surgical planning, reuse of presurgical data ("Copy & Paste"), and final impression for passive fitting of final restoration. To illustrate these principles, a case with complete-mouth implant rehabilitation completed within six visits using this virtual patient workflow is presented. CONCLUSION: The virtual patient workflow serves as an invaluable tool to perform treatment planning, enhance efficiency, and ensure predictable outcomes in esthetic complete arch implant rehabilitation. CLINICAL SIGNIFICANCE: Virtual workflows are increasingly prevalent in esthetic implant rehabilitation. Nevertheless, these workflows necessitate a distinct set of knowledge and tools divergent from conventional dentistry practices. This article offers guidelines and recommendations for dental clinicians who are new to this field.

Enhancing the Treatment of Uncontrolled Inflammation through the Targeted Delivery of TPCA-1-Loaded Nanoparticles.

Uncontrolled inflammation is a pathological state that underlies many diseases. Despite the development of numerous anti-inflammatory agents, the treatment of uncontrolled inflammation remains a challenging task. We developed a targeted delivery system for [5-(p-fluorophenyl)-2-ureido]thiophene-3-carboxamide (TPCA-1), a potent inhibitor of the NF-κB signaling pathway. The system comprises TPCA-1-loaded nanoparticles (NPs) functionalized with a monoclonal antibody (mAb) that specifically binds to the break point of the IgD6 region of the platelet/endothelial cell adhesion molecule-1 (PECAM-1) extracellular segment that is overexposed on the injured endothelium and activated macrophages during the pathogenesis of inflammation. In vitro binding and cellular uptake experiments revealed that the mAb modification on NPs could significantly enhance uptake by both Raw264.7 and HUVEC compared with unmodified NPs. In studies conducted at the cellular level focusing on anti-inflammatory and antioxidant effects, this formulation was found to effectively inhibit M1 polarization of macrophages, downregulate the secretion of pro-inflammatory cytokines, and reduce the production of reactive oxygen species (ROS) and nitric oxide (NO). In an animal model of vascular endothelial injury with acute inflammation, these NPs were capable of delivering TPCA-1 to inflammatory lesions in a targeted manner. Compared with the free agent-treated group, the NP-treated group exhibited reduced infiltration of inflammatory cells. In conclusion, our study demonstrates that this targeted delivery of TPCA-1-loaded NPs represents a promising strategy for improved mitigation of uncontrolled inflammation.

Advances in CAR-Engineered Immune Cell Generation: Engineering Approaches and Sourcing Strategies.

Chimeric antigen receptor T-cell (CAR-T) therapy has emerged as a highly efficacious treatment modality for refractory and relapsed hematopoietic malignancies in recent years. Furthermore, CAR technologies for cancer immunotherapy have expanded from CAR-T to CAR-natural killer cell (CAR-NK), CAR-cytokine-induced killer cell (CAR-CIK), and CAR-macrophage (CAR-MΦ) therapy. Nevertheless, the high cost and complex manufacturing processes of ex vivo generation of autologous CAR products have hampered broader application. There is an urgent need to develop an efficient and economical paradigm shift for exploring new sourcing strategies and engineering approaches toward generating CAR-engineered immune cells to benefit cancer patients. Currently, researchers are actively investigating various strategies to optimize the preparation and sourcing of these potent immunotherapeutic agents. In this work, the latest research progress is summarized. Perspectives on the future of CAR-engineered immune cell manufacturing are provided, and the engineering approaches, and diverse sources used for their development are focused upon.

circ_SPEF2 Regulates the Balance of Treg Cells by Regulating miR-16-5p/BACH2 in Lymphoma and Participates in the Immune Response.

BACKGROUND: This study aims to explore the potential mechanism of action of the newly discovered hsa_circ_0128899 (circSPEF2) in diffuse large B-cell lymphoma (DLBCL). METHODS: circSPEF2, miR-16-5p and BTB and CNC homologue 2 (BACH2) expression patterns in DLBCL patients and cell lines were studied by RT-qPCR. The biological function of circSPEF2 in vitro and in vivo was investigated by function acquisition experiments. The proliferation activity of lymphoma cells was detected by MTT. Bax, Caspase-3, and Bcl-2 were determined by Western Blot. Apoptosis and the ratio of CD4 to Treg of immune cells in the co-culture system were analyzed by flow cytometry. The mechanism of action of circSPEF2 in DLBCL progression was further investigated by RIP and dual luciferase reporter experiments. RESULTS: circSPEF2 was a circRNA with abnormally down-regulated expression in DLBCL. Increasing circSPEF2 expression inhibited the proliferative activity and induced apoptosis of lymphoma cells in vitro and in vivo, as well as increased CD4+T cells and decreased Treg cell proportion of immune cells in the tumor microenvironment. Mechanically, circSPEF2 was bound to miR-16-5p expression, while BACH2 was targeted by miR-16-5p. circSPEF2 overexpression-mediated effects on lymphoma progression were reversible by upregulating miR-16-5p or downregulating BACH2. CONCLUSIONS: circSPEF2 can influence DLBCL progression by managing cellular proliferation and apoptosis and the proportion of immune cells Treg and CD4 through the miR-16-5p/BACH2 axis.

Prosthetic articulator-based implant rehabilitation virtual patient: A technique bridging implant surgery and reconstructive dentistry.

This technique report describes a fully digital workflow to create a prosthetic articulator-based implant rehabilitation (PAIR) virtual patient for complete-arch or complete-mouth implant rehabilitation. This workflow uses a custom gothic arch tracer during the cone beam computed tomography (CBCT) scan and a 3-dimensional virtual facebow when superimposing data. The PAIR virtual patient possesses reliable centric relation and vertical dimension of occlusion and is compatible with virtual articulators. Computer-aided implant planning and a digital prosthetic design can be seamlessly integrated by using this virtual patient.

Soft tissue phenotype modification impacts on peri-implant stability: a comparative cohort study.

OBJECTIVES: Soft tissue phenotype modification (STPM) could be performed to maintain peri-implant health. Therefore, the aim of the study was to analyze tissue alteration around implants following soft tissue phenotype modification during implant uncovering surgery. MATERIALS AND METHODS: Patients who had STPM (either pouch roll or modified roll technique) during implant second-stage surgery with at least 12-month follow-up were included. Clinical and radiographic parameters including mucosal tissue thickness (MTT), recession (REC), keratinized mucosa width (KMW), probing pocket depth (PPD), marginal bone loss (MBL), emergence profile, and emergence angle were extracted from 2-week, 2-month, and 12-month visits after second-stage surgery. RESULTS: Twenty-eight patients with 33 implants that fulfilled the inclusion criteria were included. After soft tissue phenotype modification, at 2 weeks, REC was negatively correlated to mean MTT at mid-buccal site (r = - 0.41, p = 0.018) and borderline correlated at mid-lingual site (r = - 0.343, p = 0.051). Stable KMW was maintained from 2 weeks to 12 months with minimal shrinkage rate (3 ~ 14%). MBL change was limited (0.24 ~ 0.47 mm) after STPM. All implants had shallow PPD (≤ 3 mm) with the absence of bleeding on probing. Emergence angle at the mesial side, however, was significantly correlated to surgical techniques, which indicated pouch roll technique would have 6.96 degrees more than modified roll technique (p = 0.024). CONCLUSIONS: Soft tissue phenotype modification, either pouch roll or modified roll technique, during uncovering surgery resulted in favorable clinical outcomes. Thin mucosal tissue thickness and pouch roll technique are the factors related to more recession at 2 weeks. Pouch roll technique could influence the restorative design by having a wide emergence angle at the mesial side. CLINICAL RELEVANCE: Modified and pouch roll techniques during uncovering surgery were viable methods to yield favorable peri-implant health, while the preciseness of pouch roll technique was required to avoid mucosal recession and inadequate restorative design.