A Comprehensive Review on Silk Fibroin as a Persuasive Biomaterial for Bone Tissue Engineering.

Bone tissue engineering (BTE) utilizes a special mix of scaffolds, cells, and bioactive factors to regulate the microenvironment of bone regeneration and form a three-dimensional bone simulation structure to regenerate bone tissue. Silk fibroin (SF) is perhaps the most encouraging material for BTE given its tunable mechanical properties, controllable biodegradability, and excellent biocompatibility. Numerous studies have confirmed the significance of SF for stimulating bone formation. In this review, we start by introducing the structure and characteristics of SF. After that, the immunological mechanism of SF for osteogenesis is summarized, and various forms of SF biomaterials and the latest development prospects of SF in BTE are emphatically introduced. Biomaterials based on SF have great potential in bone tissue engineering, and this review will serve as a resource for future design and research.

Hypoxia-Inducible Factors Signaling in Osteogenesis and Skeletal Repair.

Sufficient oxygen is required to maintain normal cellular and physiological function, such as a creature's development, breeding, and homeostasis. Lately, some researchers have reported that both pathological hypoxia and environmental hypoxia might affect bone health. Adaptation to hypoxia is a pivotal cellular event in normal cell development and differentiation and in pathological settings such as ischemia. As central mediators of homeostasis, hypoxia-inducible transcription factors (HIFs) can allow cells to survive in a low-oxygen environment and are essential for the regulation of osteogenesis and skeletal repair. From this perspective, we summarized the role of HIF-1 and HIF-2 in signaling pathways implicated in bone development and skeletal repair and outlined the molecular mechanism of regulation of downstream growth factors and protein molecules such as VEGF, EPO, and so on. All of these present an opportunity for developing therapies for bone regeneration.

Finite-Time Dynamic Tracking Control of Parallel Robots with Uncertainties and Input Saturation.

This paper considers the finite-time dynamic tracking control for parallel robots with uncertainties and input saturation via a finite-time nonsingular terminal sliding mode control scheme. A disturbance observer is designed to estimate the lumped disturbance in the dynamic model of the parallel robot, including modeling errors, friction and external disturbance. By introducing the fractional exponential powers into the existing asymptotic convergent auxiliary system, a novel finite-time convergent auxiliary system is constructed to compensate for input saturation. The finite-time nonsingular terminal sliding mode control is proposed based on the disturbance estimation and the state of the novel auxiliary system, so that the convergence performance, control accuracy and robustness are improved. Due to the estimation and compensation for the lumped disturbance, the inherent chattering characteristic of sliding mode control can be alleviated by reducing the control gain. The finite-time stability of the closed-loop system is proved with Lyapunov theory. Finally, simulation and experimental research on the dynamic control of a conveying parallel robot are carried out to verify the effectiveness of the proposed method.

Class I-restricted T-cell responses to a polymorphic peptide in a gene therapy clinical trial for α-1-antitrypsin deficiency.

Adeno-associated virus (AAV)-mediated gene therapy is currently being pursued as a treatment for the monogenic disorder α-1-antitrypsin (AAT) deficiency. Results from phase I and II studies have shown relatively stable and dose-dependent increases in transgene-derived wild-type AAT after local intramuscular vector administration. In this report we describe the appearance of transgene-specific T-cell responses in two subjects that were part of the phase II trial. The patient with the more robust T-cell response, which was associated with a reduction in transgene expression, was characterized more thoroughly in this study. We learned that the AAT-specific T cells in this patient were cytolytic in phenotype, mapped to a peptide in the endogenous mutant AAT protein that contained a common polymorphism not incorporated into the transgene, and were restricted by a rare HLA class I C alleles present only in this patient. These human studies illustrate the genetic influence of the endogenous gene and HLA haplotype on the outcome of gene therapy.

Increased mucosal CD4+ T cell activation in rhesus macaques following vaccination with an adenoviral vector.

UNLABELLED: The possibility that vaccination with adenovirus (AdV) vectors increased mucosal T cell activation remains a central hypothesis to explain the potential enhancement of HIV acquisition within the Step trial. Modeling this within rhesus macaques is complicated because human adenoviruses, including human adenovirus type 5 (HAdV-5), are not endogenous to macaques. Here, we tested whether vaccination with a rhesus macaque-derived adenoviral vector (simian adenovirus 7 [SAdV-7]) enhances mucosal T cell activation within rhesus macaques. Following intramuscular SAdV-7 vaccination, we observed a pronounced increase in SAdV-7-specific CD4(+) T cell responses in peripheral blood and, more dramatically, in rectal mucosa tissue. Vaccination also induced a significant increase in the frequency of activated memory CD4(+) T cells in SAdV-7- and HAdV-5-vaccinated animals in the rectal mucosa but not in peripheral blood. These fluctuations within the rectal mucosa were also associated with a pronounced decrease in the relative frequency of naive resting CD4(+) T cells. Together, these results indicate that peripheral vaccination with an AdV vector can increase the activation of mucosal CD4(+) T cells, potentially providing an experimental model to further evaluate the role of host-vector interactions in increased HIV acquisition after AdV vector vaccination. IMPORTANCE: The possibility that vaccination with a human adenovirus 5 vector increased mucosal T cell activation remains a central hypothesis to explain the potential enhancement of human immunodeficiency virus (HIV) acquisition within the Step trial. In this study, we tested whether vaccination with a rhesus macaque-derived adenoviral vector in rhesus macaques enhances mucosal CD4(+) T cell activation, the main cell target of simian immunodeficiency virus (SIV)/HIV. The results showed that vaccination with an adenoviral vector indeed increases activation of mucosal CD4(+) T cells and potentially increases susceptibility to SIV infection.

LIPUS as a potential strategy for periodontitis treatment: A review of the mechanisms.

Periodontitis is a chronic inflammatory condition triggered by oral bacteria. A sustained inflammatory state in periodontitis could eventually destroy the alveolar bone. The key objective of periodontal therapy is to terminate the inflammatory process and reconstruct the periodontal tissues. The traditional Guided tissue regeneration (GTR) procedure has unstable results due to multiple factors such as the inflammatory environment, the immune response caused by the implant, and the operator's technique. Low-intensity pulsed ultrasound (LIPUS), as acoustic energy, transmits the mechanical signals to the target tissue to provide non-invasive physical stimulation. LIPUS has positive effects in promoting bone regeneration, soft-tissue regeneration, inflammation inhibition, and neuromodulation. LIPUS can maintain and regenerate alveolar bone during an inflammatory state by suppressing the expression of inflammatory factors. LIPUS also affects the cellular behavior of periodontal ligament cells (PDLCs), thereby protecting the regenerative potential of bone tissue in an inflammatory state. However, the underlying mechanisms of the LIPUS therapy are still yet to be summarized. The goal of this review is to outline the potential cellular and molecular mechanisms of periodontitis-related LIPUS therapy, as well as to explain how LIPUS manages to transmit mechanical stimulation into the signaling pathway to achieve inflammatory control and periodontal bone regeneration.

3D printed reduced graphene oxide-GelMA hybrid hydrogel scaffolds for potential neuralized bone regeneration.

Peripheral nerves participate in bone growth and repair by secreting neurotransmitters, and enable new bone to possess physiological bone-sensing capability. However, it is difficult to achieve synchronized nerve regeneration during the healing process of large bone defects at present. As a bioactive nanomaterial, reduced graphene oxide (rGO) can promote neuronal differentiation and myelination of Schwann cells (SCs), while enhancing the adhesion and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) through its strong non-covalent binding ability. In this study, 3D printing-based rGO/GelMA hydrogels with enhanced osteogenic and neurogenic dual differentiation were used to simultaneously load SCs and BMSCs. By changing the concentration of rGO(0.03%/0.05%/0.1%), the compressive strength, rheological properties and aperture of the hydrogel can be improved. In vitro, cell live/death staining, phalloidin staining and SEM showed that cells loaded on the hydrogel had a high survival rate (85%) and good adhesion ability. In vivo, we found that the rGO/GelMA hydrogel exhibited the same low inflammatory response compared to the pure-GelMA group and the cell-only group, but surrounded by collagen fibers. Meanwhile, the osteogenic and neural proteins in the rGO/GelMA group were found to be highly expressed in immunohistochemistry and immunofluorescence. In this study, a scaffold material containing double cells was used to promote synergistic regeneration of nerves and bone, providing a promising strategy for the preparation of personalized and functionalized biomimetic bone material.

Correction: Highly efficient photothermal branched Au-Ag nanoparticles containing procyanidins for synergistic antibacterial and anti-inflammatory immunotherapy.

Correction for 'Highly efficient photothermal branched Au-Ag nanoparticles containing procyanidins for synergistic antibacterial and anti-inflammatory immunotherapy' by Hanchi Wang et al., Biomater. Sci., 2023, https://doi.org/10.1039/d2bm01212j.

Highly efficient photothermal branched Au-Ag nanoparticles containing procyanidins for synergistic antibacterial and anti-inflammatory immunotherapy.

Periodontitis is an inflammatory disease caused by bacterial infection. Excessive immune response and high levels of reactive oxygen species (ROS) further lead to the irreversible destruction of surrounding tissues. Developing new antimicrobial materials that regulate the immune system to resist inflammation can effectively treat periodontal inflammation. A nanoplatform integrating Ag+, photothermal therapy (PTT), and procyanidins (PC) for precision antibacterial and synergistic immunotherapy for periodontitis was proposed. This work loaded PC into AuAg nanoparticles, and the resulting nanocomposite was named AuAg-PC. PTT can effectively remove pathogenic bacteria, but high temperatures can cause tissue damage. Ag+ contributes to the preparation of a nanoparticle branched structure that improves the photothermal efficiency and helps PTT achieve an excellent antibacterial effect and avoid periodontal tissue damage. PC regulates host immunity by eliminating intracellular ROS, inhibiting inflammatory factors, and regulating macrophage polarisation in periodontal disease sites. It enhances the host's resistance to bacterial inflammation. AuAg-PC exerted an excellent anti-inflammatory effect and promoted tissue repair in animal periodontal inflammation models. Hence, AuAg-PC significantly combats periodontal pathogens and shows great application potential in the photothermal-assisted immunotherapy of periodontitis. This design provided a new controllable and efficient treatment platform for controlling persistent inflammation infection and regulating immunity.

Application of Bone Marrow-Derived Macrophages Combined with Bone Mesenchymal Stem Cells in Dual-Channel Three-Dimensional Bioprinting Scaffolds for Early Immune Regulation and Osteogenic Induction in Rat Calvarial Defects.

The host immune response to biomaterials is critical for determining scaffold fate and bone regeneration outcomes. Three-dimensional (3D) bioprinted scaffolds encapsulated with living cells can improve the inflammatory microenvironment and further accelerate bone repair. Here, we screened and adopted 8% methacrylamidated gelatin (GelMA)/1% methacrylamidated hyaluronic acid (HAMA) as the encapsulation system for rat bone marrow-derived macrophages (BMMs) and 3% Alginate/0.5 mg/mL graphene oxide (GO) as the encapsulation system for rat bone mesenchymal stem cells (BMSCs), thus forming a dual-channel bioprinting scaffold. The 8% GelMA/1% HAMA/3% Alginate/0.5 mg/mL GO (8/1/3/0.5) group could form a scaffold with a stable structure, good mechanical properties, and satisfied biocompatibility. When exploring the crosstalk between BMMs and BMSCs in vitro, we found that BMSCs could promote the polarization of BMMs to M2 type at the early stage, reduce the pro-inflammatory gene expression, and increase anti-inflammatory gene expression; conversely, BMMs can promote the osteogenic differentiation of BMSCs. In addition, in the model of rat calvarial defects, the dual-channel scaffold encapsulated with BMMs and BMSCs was more effective than the single-cell scaffold and the acellular scaffold. The paracrine of BMMs and BMSCs in the biodegradable dual-channel scaffold effectively promoted the M2-type polarization of macrophages in the microenvironment of early bone defects, avoided excessive inflammatory responses, and further promoted bone repair. In conclusion, our findings suggested that using 3D bioprinting to simultaneously encapsulate two primary cells of BMMs and BMSCs in a dual-channel system may be an effective way to promote bone repair from the perspective of early immune regulation and late induction of osteogenesis.

Susceptibility to SIV Infection After Adenoviral Vaccination in a Low Dose Rhesus Macaque Challenge Model.

BACKGROUND: Vaccination with the Merck human adenovirus serotype-5 (HAdV-5) vectored HIV-1 subtype B gag/pol/nef vaccine was unexpectedly associated with enhanced susceptibility to HIV-1 infection in uncircumcised HAdV-5 seropositive men. It has been hypothesized that vaccination may have resulted in activated CD4+ T lymphocytes trafficking to mucosal sites thereby increasing targets for HIV infection. We have previously shown that AdV-vector vacci-nation in rhesus macaques resulted in an increase in the frequency of activated mucosal CD4+ T cells. However, whether this increase in activation is sufficient to increase susceptibility to HIV/SIV infection is unclear. METHODS: To examine this scenario, we developed a preliminary, proof-of-concept vaccination-challenge model in order to examine vaccine-induced SIV susceptibility in rhesus macaques. Rhesus macaques (n = 10/group) were vaccinated with a simian AdV-7 (SAdV-7)-vector encoding an irrelevant insert (SARS spike) and challenged 5 weeks post-prime in an escalating dosing regimen starting with sub-infectious doses (1:10,000 or 2TCID50) of SIVmac251. RESULTS: In contrast to our previous study, the SAdV-7 vaccine regimen did not induce detectable mucosal CD4+ T cell activation at the time points assessed in animals obtained from a different vendor and housed in a different facility. Within the power of the study, we did not observe significantly increased SIV acquisition in SAdV-7-vaccinated (5/10) versus placebo-vaccinated (3/10) macaques after repeated low-dose intra-rectal SIVmac251 challenge (P < 0.2). CONCLUSIONS: These results lay groundwork for future experiments to assess vaccine-induced SIV susceptibility in rhesus macaques. Further larger-scale studies are necessary to confirm the AdV-vector vaccination associated trend towards increased SIV/HIV acquisition and clarify associated mechanisms.

Intramuscular administration of AAV overcomes pre-existing neutralizing antibodies in rhesus macaques.

The seroprevalence of neutralizing antibodies (NAbs) to adeno-associated viral (AAV) vector capsids may preclude a percentage of the population from receiving gene therapy, particularly following systemic vector administration. We hypothesized that the use of intramuscular (IM) administration of AAV vectors might circumvent this issue. IM injections were used to administer AAV8 vectors expressing either secreted or non-secreted transgenes into mice and the influence of NAbs supplied by pre-administration of pooled human IgG on transgene expression was evaluated. We then studied the impact of naturally occurring pre-existing AAV8 NAbs on expression of a secreted transgene following IM vector delivery in rhesus macaques. Finally, we evaluated the ability to readminister AAV vectors via IM injections in rhesus macaques. In mice, the presence of AAV8 NAbs had no effect on gene expression in the injected skeletal muscle. However, liver transgene expression following hepatic distribution of the vector was ablated. In rhesus macaques, naturally occurring pre-existing AAV8 NAb titers of ⩽1:160 had no effect on expression levels of a secreted transgene after IM delivery of the vector. Additionally, readministration of AAV vectors was possible by IM injection into the previously injected muscle groups, with no effect on transgene expression by the original vector. Therefore, the presence of pre-existing NAbs in the human population should not preclude subjects from receiving gene therapy by IM administration of the vector so long as sufficient levels of secreted transgene expression can be produced without the involvement of liver.

Preexisting Neutralizing Antibodies to Adeno-Associated Virus Capsids in Large Animals Other Than Monkeys May Confound In Vivo Gene Therapy Studies.

Adeno-associated virus (AAV) vectors are currently being tested not only in small animal models such as mice but also in large animal models, including pigs, dogs, and horses. Natural exposure to AAV occurs in most of the species used in these studies and potentially elicits a neutralizing humoral immune response to AAV. In this study, we show the prevalence of neutralizing antibodies (NAbs) to several AAV serotypes in these large animals as measured by an in vitro NAb assay and the ability of these NAbs to block AAV transduction in an in vivo mouse model of NAb passive transfer assay. The results of this study clearly show the importance of evaluating large animal models for the presence of AAV NAbs before enrolling them in AAV-mediated gene therapy studies.