Targeted drug delivery of engineered mesenchymal stem/stromal-cell-derived exosomes in cardiovascular disease: recent trends and future perspectives.

In recent years, stem cells and their secretomes, notably exosomes, have received considerable attention in biomedical applications. Exosomes are cellular secretomes used for intercellular communication. They perform the function of intercellular messengers by facilitating the transport of proteins, lipids, nucleic acids, and therapeutic substances. Their biocompatibility, minimal immunogenicity, targetability, stability, and engineerable characteristics have additionally led to their application as drug delivery vehicles. The therapeutic efficacy of exosomes can be improved through surface modification employing functional molecules, including aptamers, antibodies, and peptides. Given their potential as targeted delivery vehicles to enhance the efficiency of treatment while minimizing adverse effects, exosomes exhibit considerable promise. Stem cells are considered advantageous sources of exosomes due to their distinctive characteristics, including regenerative and self-renewal capabilities, which make them well-suited for transplantation into injured tissues, hence promoting tissue regeneration. However, there are notable obstacles that need to be addressed, including immune rejection and ethical problems. Exosomes produced from stem cells have been thoroughly studied as a cell-free strategy that avoids many of the difficulties involved with cell-based therapy for tissue regeneration and cancer treatment. This review provides an in-depth summary and analysis of the existing knowledge regarding exosomes, including their engineering and cardiovascular disease (CVD) treatment applications.

PCAT-1' s role in wound healing impairment: Mitochondrial dysfunction and bone marrow stem cell differentiation inhibition via PKM2/ß-catenin pathway and its impact on implant osseo-integration.

This study focused on unravelling the role of PCAT-1 in wound-healing process, particularly its impact on regenerative and osteogenic abilities of mesenchymal stem cells (MSCs). We delved into how PCAT-1 regulates mitochondrial oxidative phosphorylation (OXPHOS) and interacts with pivotal molecular pathways, especially ß-catenin and PKM2, using human bone marrow-derived MSCs. MSCs were cultured under specific conditions and PCAT-1 expression was modified through transfection. We thoroughly assessed several critical parameters: MSC proliferation, mitochondrial functionality, ATP production and expression of wound healing and osteogenic differentiation markers. Further, we evaluated alkaline phosphatase (ALP) activity and mineral deposition, essential for bone healing. Our findings revealed that overexpressing PCAT-1 significantly reduced MSC proliferation, hampered mitochondrial performance and lowered ATP levels, suggesting the clear inhibitory effect of PCAT-1 on these vital wound-healing processes. Additionally, PCAT-1 overexpression notably decreased ALP activity and calcium accumulation in MSCs, crucial for effective bone regeneration. This overexpression also led to the reduction in osteogenic marker expression, indicating suppression of osteogenic differentiation, essential in wound-healing scenarios. Moreover, our study uncovered a direct interaction between PCAT-1 and the PKM2/ß-catenin pathway, where PCAT-1 overexpression intensified PKM2 activity while inhibiting ß-catenin, thereby adversely affecting osteogenesis. This research thus highlights PCAT-1's significant role in impairing wound healing, offering insights into the molecular mechanisms that may guide future therapeutic strategies for enhancing wound repair and bone regeneration.

The role of extracellular vesicles in periodontitis: pathogenesis, diagnosis, and therapy.

Periodontitis is a prevalent disease and one of the leading causes of tooth loss. Biofilms are initiating factor of periodontitis, which can destroy periodontal tissue by producing virulence factors. The overactivated host immune response is the primary cause of periodontitis. The clinical examination of periodontal tissues and the patient's medical history are the mainstays of periodontitis diagnosis. However, there is a lack of molecular biomarkers that can be used to identify and predict periodontitis activity precisely. Non-surgical and surgical treatments are currently available for periodontitis, although both have drawbacks. In clinical practice, achieving the ideal therapeutic effect remains a challenge. Studies have revealed that bacteria produce extracellular vesicles (EVs) to export virulence proteins to host cells. Meanwhile, periodontal tissue cells and immune cells produce EVs that have pro- or anti-inflammatory effects. Accordingly, EVs play a critical role in the pathogenesis of periodontitis. Recent studies have also presented that the content and composition of EVs in saliva and gingival crevicular fluid (GCF) can serve as possible periodontitis diagnostic indicators. In addition, studies have indicated that stem cell EVs may encourage periodontal regeneration. In this article, we mainly review the role of EVs in the pathogenesis of periodontitis and discuss their diagnostic and therapeutic potential.

Ultrasound Responsive Smart Implantable Hydrogels for Targeted Delivery of Drugs: Reviewing Current Practices.

Over the last two decades, the process of delivering therapeutic drugs to a patient with a controlled release profile has been a significant focus of drug delivery research. Scientists have given tremendous attention to ultrasound-responsive hydrogels for several decades. These smart nanosystems are more applicable than other stimuli-responsive drug delivery vehicles (ie UV-, pH- and thermal-, responsive materials) because they enable more efficient targeted treatment via relatively non-invasive means. Ultrasound (US) is capable of safely transporting energy through opaque and complex media with minimal loss of energy. It is capable of being localized to smaller regions and coupled to systems operating at various time scales. However, the properties enabling the US to propagate effectively in materials also make it very difficult to transform acoustic energy into other forms that may be used. Recent research from a variety of domains has attempted to deal with this issue, proving that ultrasonic effects can be used to control chemical and physical systems with remarkable specificity. By obviating the need for multiple intravenous injections, implantable US responsive hydrogel systems can enhance the quality of life for patients who undergo treatment with a varied dosage regimen. Ideally, the ease of self-dosing in these systems would lead to increased patient compliance with a particular therapy as well. However, excessive literature has been reported based on implanted US responsive hydrogel in various fields, but there is no comprehensive review article showing the strategies to control drug delivery profile. So, this review was aimed at discussing the current strategies for controlling and targeting drug delivery profiles using implantable hydrogel systems.

Nerve growth factor enhances the therapeutic effect of mesenchymal stem cells on diabetic periodontitis.

Patients with diabetes frequently suffer from periodontitis, which progresses rapidly and is difficult to cure. Mesenchymal stem cell (MSC) transplantation may effectively treat periodontitis, but high glucose limits its therapeutic effect in diabetes. Nerve growth factor (NGF) has the functions of cell protection, anti-apoptosis and immune regulation, and may have potential application in diabetic periodontitis. In the present study, flow cytometry indicated that NGF inhibited MSC apoptosis induced by high glucose. Of note, high glucose promoted the transformation of MSCs into the proinflammatory type. NGF inhibited this transformation of MSCs under diabetic conditions and further decreased the proportion of T cells and monocytes/macrophages among lymphocytes. An animal model of diabetic periodontitis was constructed and MSC transplantation was demonstrated to reduce alveolar bone loss caused by diabetes. NGF enhanced the therapeutic effect of MSCs and maintained transplanted MSC survival in periodontal tissue of diabetic mice. Immunohistochemical analysis of periodontal tissues suggested that in the NGF group, infiltration of T cells and macrophages was reduced. Neurotrophic receptor tyrosine kinase 1 was indicated to have a key role in these effects of NGF. In conclusion, NGF may enhance the therapeutic effect of MSCs on diabetic periodontitis by protecting the cells and promoting the transformation of MSCs into the immunosuppressive type.

Vorapaxar stabilizes permeability of the endothelial barrier under cholesterol stimulation via the AKT/JNK and NF­κB signaling pathways.

Atherosclerosis (AS) is an inflammatory disease that occurs in the arterial wall and is characterized by progressive lipid accumulation within the intima of large arteries, leading to the dysfunction of endothelial cells and further destruction of the endothelial barrier and vascular tone. Arterial intima injury accelerates the adhesion and activation of platelets at the injury site. The activation of platelets results in the secretion of growth factors, leading to the migration and proliferation of vascular smooth muscle cells (VSMCs), promoting the formation of plaque, resulting in the formation of thrombus. The present study found that vorapaxar could alleviate the inflammatory response induced by a high concentration of cholesterol stimulation and increase the release of nitric oxide (NO) via the protein kinase B (AKT) signaling pathway and regulation of the intracellular concentration of Ca2+ ([Ca2+]i). We also found that vorapaxar could reduce the damage of DNA caused by cholesterol stimulation and regulate the cell cycle via the AKT/JNK signaling pathway and its downstream molecules glycogen synthase kinase 3ß (GSK­3ß) and connexin 43, maintaining the integrity of the endothelial barrier and proliferation of endothelial cells, serving a protective role in endothelial cells.

Fracture behaviors of maxillary central incisors with flared root canals restored with CAD/CAM integrated glass fiber post-and-core.

The objective was to evaluate the fracture resistance properties of maxillary incisors with flared canals restored with computer aided design and computer aided manufacture (CAD/CAM) integrated glass fiber post-and-core. Thirty prepared flared root canals were selected in vitro and restored with CAD/CAM integrated fiber post-and-core (Group A), prefabricated fiber posts (Group B), and cast gold alloy (Group C), respectively. After submitted to fatigue loading, each specimen was subjected to a static loading until fracture. Analysis of variance (ANOVA) tests were used to determine statistical differences. The mean fracture strengths of Groups A and C were significantly higher than those of Group B, whereas no differences were observed between Groups A and C. In addition, reparable fracture modes were mostly observed in Group A while irreparable and catastrophic fractures were mostly found in Groups B and C. These results demonstrate that, in comparison to traditional treatments, CAD/CAM integrated glass fiber post-and-core restoration significantly enhances the fracture resistance of flared root canals.

Isolation and multiple differentiation potential assessment of human gingival mesenchymal stem cells.

The aim of this study was to isolate human mesenchymal stem cells (MSCs) from the gingiva (GMSCs) and confirm their multiple differentiation potentials, including the odontogenic lineage. GMSCs, periodontal ligament stem cells (PDLSCs) and dermal stem cells (DSCs) cultures were analyzed for cell shape, cell cycle, colony-forming unit-fibroblast (CFU-F) and stem cell markers. Cells were then induced for osteogenic and adipogenic differentiation and analyzed for differentiation markers (alkaline phosphatase (ALP) activity, mineralization nodule formation and Runx2, ALP, osteocalcin (OCN) and collagen I expressions for the osteogenic differentiation, and lipid vacuole formation and PPARγ-2 expression for the adipogenic differentiation). Besides, the odontogenic differentiation potential of GMSCs induced with embryonic tooth germ cell-conditioned medium (ETGC-CM) was observed. GMSCs, PDLSCs and DSCs were all stromal origin. PDLSCs showed much higher osteogenic differentiation ability but lower adipogenic differentiation potential than DSCs. GMSCs showed the medial osteogenic and adipogenic differentiation potentials between those of PDLSCs and DSCs. GMSCs were capable of expressing the odontogenic genes after ETGC-CM induction. This study provides evidence that GMSCs can be used in tissue engineering/regeneration protocols as an approachable stem cell source.

Construction of tissue-engineered osteochondral composites and repair of large joint defects in rabbit.

In this study, a novel three-dimensional (3D) heterogeneous/bilayered scaffold was constructed to repair large defects in rabbit joints. The scaffold includes two distinct but integrated layers corresponding to the cartilage and bone components. The upper layer consists of gelatin, chondroitin sulphate and sodium hyaluronate (GCH), and the lower layer consists of gelatin and ceramic bovine bone (GCBB). The two form a 3D bilayered scaffold (GCH-GCBB), which mimics the natural osteochondral matrix for use as a scaffold for osteochondral tissue engineering. The purpose of this study was to evaluate the efficacy of this novel scaffold, combined with chondrocytes and bone marrow stem cells (BMSCs) to repair large defects in rabbit joints. Thirty-six large defects in rabbit femoral condyles were created; 12 defects were treated with the same scaffold combined with cells (group A); another 12 defects were treated with cell-free scaffolds (group B); the others were untreated (group C). At 6 and 12 weeks, in group A hyaline-like cartilage formation could be observed by histological examination; the newly formed cartilage, which stained for type II collagen, was detected by RT-PCR at high-level expression. Most of the GCBB was replaced by bone, while little remained in the underlying cartilage. At 36 weeks, GCBB was completely resorbed and a tidemark was observed in some areas. In contrast, groups B and C showed no cartilage formation but a great amount of fibrous tissue, with only a little bone formation. In summary, this study demonstrated that a novel scaffold, comprising a top layer of GCH, having mechanical properties comparable to native cartilage, and a bottom layer composed of GCBB, could be used to repair large osteochondral defects in joints.

Association of three bacterial species and periodontal status in Chinese adults: an epidemiological approach.

Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythia are oral pathogens associated with periodontitis. The association between these three bacteria and periodontal disease has been reported in populations of many countries. However, corresponding data in Chinese populations are still lacking. The aim of this study was to detect these pathogens in subgingival plaque collected from 468 subjects with chronic periodontitis in a group of Chinese adults by using a PCR method and to determine the degree of association between the target bacteria and periodontal status based on logistic regression analysis. A. actinomycetemcomitans, P. gingivalis, and T. forsythia were found in 20.5%, 70.7%, and 77.1% of the subjects, respectively. About one-third (36.1%) of subjects had chronic periodontitis. Upon univariate analysis, age, male gender, current smoking status, diabetes, and the presence of A. actinomycetemcomitans or P. gingivalis were positively associated with chronic periodontitis, whereas education and income exhibited inverse associations with chronic periodontitis. Upon multivariate analysis, education, current smoking status, diabetes, and the presence of A. actinomycetemcomitans and P. gingivalis remained significant. The adjusted odds ratios for having chronic periodontitis were 2.5 and 3.4 in subjects positive for A. actinomycetemcomitans and P. gingivalis, respectively. However, no significant association was observed between the presence of T. forsythia and periodontal status. This study assesses the prevalence of periodontal pathogens and chronic periodontitis and the associations with sociodemographic characteristics among this group of Chinese adults. These findings also suggest that PCR should be considered for field oral epidemiologic studies and may be necessary in investigations presenting major logistic challenges.

Upregulation of dentin matrix protein 1 promoter activities by core binding factor alpha1 in human dental pulp stem cells.

An osteoblast-specific transcription factor, core binding factor alpha1 (Cbfalpha1), is critical for osteoblast and odontoblast differentiation. In this study, the role of Cbfalpha1 in human dentin matrix protein1 (DMP1) gene expression in human dental pulp stem cells (HDPSCs) was investigated. The desired promoter fragments were obtained and cloned into the pGL3-basic vector. It was found that Cbfalpha1 isoforms were predominantly expressed in the cytoplasm of the HDPSCs and reached to the maximum after transfection for 48h. Furthermore, forced overexpression of Cbfalpha1 induced the increase of the luciferase activities of pGL3-P1-6, especially those of pGL3-P(-505to+86) (p<0.05) were the most significant. Then the site-directed mutagenesis of Cbfalpha1 binding sites in the promoter region of nt -505 to +86 resulted in a marked decline of luciferase activities. Thus, our results suggest that Cbfalpha1 upregulates DMP1 gene expression differentially that may contribute to the spatial-temporal expression pattern of DMP1 during odontoblast differentiation.

Protective immunity in rats by intranasal immunization with Streptococcus mutans glucan-binding protein D encapsulated into chitosan-coated poly(lactic-co-glycolic acid) microspheres.

Recombinant Streptococcus mutans glucan-binding protein D (rGbpD) was incorporated into poly(lactic-co-glycolic acid) (PLGA) microspheres which then were surface-coated with chitosan. The microspheres, with a mean diameter of ca. 1.8 microm, were intranasally administered in rats. There were elevated salivary immunoglobulin A and serum immunoglobulin G antibody responses to rGbpD, as well as lower molar caries scores in immunized animals as compared to sham immunized ones. The chitosan-coated PLGA microspheres are thus potentially useful for antigen delivery in dental caries vaccination.

[Comparison and analysis of human dentin matrix protein 1 promoter activity in three different cells].

OBJECTIVE: To observe and compare the luciferase activities of different length segments of human dentin matrix protein 1 promoter in human dental pulp stem cells (HDPSC), osteoblasts (OC) and Hela cells. METHODS: The differentlength desired DNA segments were obtained from 2 195 bp Dmp1 promoter cloned by PCR method. The amplified promoter segments with different length were cloned into luciferase report gene vector pGL3-Basic, the correct orientation of those inserts was verified by cutting with two different restrict enzymes. The luciferase activity was observed after different pGL3-PDmp1 vectors were transfected transiently into those three different-type cells. RESULTS: 6 Dmp1 promoter segments with different-length were obtained successfully, and luciferase report gene vectors with different promoter segments were successfully constructed after identified by restriction enzymes cutting. They had different luciferase activities when they were transfected transiently into HDPSC, and the region of -505(-)-193 bp and -935(-)-505 bp could be regarded as the specific promoters of Dmp1 promoter for HDPSC and OC respectively, which could include the basic regulatory elements. CONCLUSION: The correct clone of the upstream of human Dmp1 promoter segments with different length had been obtained, and they had strong luciferase activities in HDPSC and OC, but very low in Hela cell. These results will make an important basis for studying mineralized tissue-specific transcriptional regulation mechanisms of Dmp1.