3D bioprinting of DPSCs with GelMA hydrogel of various concentrations for bone regeneration.

Bioprinting technology promotes innovation of fabricating tissue engineered constructs. Dental pulp stem cells (DPSCs) have significant advantages over classical bone mesenchymal stem cells (BMSCs) and are a promising seed cell candidate for bone engineering bioprinting. However, current reports about bioprinted DPSCs for bone regeneration are incomprehensive. The objective of this study was to investigate the osteogenic potential of DPSCs in methacrylate gelatin (GelMA) hydrogels bioprinted scaffolds in vitro and in vivo. Firstly, we successfully bioprinted GelMA with different concentrations embedded with or without DPSCs. Printability, physical features and biological properties of the bioprinted constructs were evaluated. Then, osteogenic differentiation levels of DPSCs in bioprinted constructs with various concentrated GelMA were compared. Finally, effects of bioprinted constructs on cranial bone regeneration were evaluated in vivo. The results of our study demonstrated that 10% GelMA had higher compression modulus, smaller pores, lower swelling and degradation rate than 3% GelMA. Twenty-eight days after printing, DPSCs in three groups of bioprinted structures still maintained high cell activities (>90%). Moreover, DPSCs in 10% GelMA showed an upregulated expression of osteogenic markers and a highly activated ephrinB2/EphB4 signaling, a signaling involved in bone homeostasis. In vivo experiments showed that DPSCs survived at a higher rate in 10% GelMA, and more new bones were observed in DPSC-laden 10% GelMA group, compared with GelMA of other concentrations. In conclusion, bioprinted DPSC-laden 10% GelMA might be more appropriate for bone regeneration application, in contrast to GelMA with other concentrations.

An Injectable Hydrogel Loaded with GMSCs-Derived Neural Lineage Cells Promotes Recovery after Stroke.

Ischemic stroke is a devastating medical condition with poor prognosis due to the lack of effective treatment modalities. Transplantation of human neural stem cells or primary neural cells is a promising treatment approach, but this is hindered by limited suitable cell sources and low in vitro expansion capacity. This study aimed (1) use small molecules (SM) to reprogram gingival mesenchymal stem cells (GMSCs) commitment to the neural lineage cells in vitro, and (2) use hyaluronic acid (HA) hydrogel scaffolds seeded with GMSCs-derived neural lineage cells to treat ischemic stroke in vivo. Neural induction was carried out with a SM cocktail-based one-step culture protocol over a period of 24 h. The induced cells were analyzed for expression of neural markers with immunocytochemistry and quantitative real-time polymerase chain reaction (qRT-PCR). The Sprague-Dawley (SD) rats (n = 100) were subjected to the middle cerebral artery occlusion (MCAO) reperfusion ischemic stroke model. Then, after 8 days post-MCAO, the modeled rats were randomly assigned to six study groups (n = 12 per group): (1) GMSCs, (2) GMSCs-derived neural lineage cells, (3) HA and GMSCs-derived neural lineage cells, (4) HA, (5) PBS, and (6) sham transplantation control, and received their respective transplantation. Evaluation of post-stroke recovery were performed by behavioral tests and histological assessments. The morphologically altered nature of neural lineages has been observed of the GMSCs treated with SMs compared to the untreated controls. As shown by the qRT-PCR and immunocytochemistry, SMs further significantly enhanced the expression level of neural markers of GMSCs as compared with the untreated controls (all p < 0.05). Intracerebral injection of self-assembling HA hydrogel carrying GMSCs-derived neural lineage cells promoted the recovery of neural function and reduced ischemic damage in rats with ischemic stroke, as demonstrated by histological examination and behavioral assessments (all p < 0.05). In conclusion, the SM cocktail significantly enhanced the differentiation of GMSCs into neural lineage cells. The HA hydrogel was found to facilitate the proliferation and differentiation of GMSCs-derived neural lineage cells. Furthermore, HA hydrogel seeded with GMSCs-derived neural lineage cells could promote tissue repair and functional recovery in rats with ischemic stroke and may be a promising alternative treatment modality for stroke.

Strategies of functionalized GelMA-based bioinks for bone regeneration: Recent advances and future perspectives.

Gelatin methacryloyl (GelMA) hydrogels is a widely used bioink because of its good biological properties and tunable physicochemical properties, which has been widely used in a variety of tissue engineering and tissue regeneration. However, pure GelMA is limited by the weak mechanical strength and the lack of continuous osteogenic induction environment, which is difficult to meet the needs of bone repair. Moreover, GelMA hydrogels are unable to respond to complex stimuli and therefore are unable to adapt to physiological and pathological microenvironments. This review focused on the functionalization strategies of GelMA hydrogel based bioinks for bone regeneration. The synthesis process of GelMA hydrogel was described in details, and various functional methods to meet the requirements of bone regeneration, including mechanical strength, porosity, vascularization, osteogenic differentiation, and immunoregulation for patient specific repair, etc. In addition, the response strategies of smart GelMA-based bioinks to external physical stimulation and internal pathological microenvironment stimulation, as well as the functionalization strategies of GelMA hydrogel to achieve both disease treatment and bone regeneration in the presence of various common diseases (such as inflammation, infection, tumor) are also briefly reviewed. Finally, we emphasized the current challenges and possible exploration directions of GelMA-based bioinks for bone regeneration.

The Effect of Differentiation of Human Dental Pulp Stem Cells to Glial Cells on the Sensory Nerves of the Dental Pulp.

Regeneration of sensory nerves is challenging in dental pulp regeneration. Schwann cells (SCs) are essential glial cells conducive to regenerating sensory nerve, but their source is scarce. The aim of the protocol was to investigate the regenerative potential of Schwann-like cells derived from dental pulp stem cells (SC-DPSCs) for sensory nerve regrowth. SC-DPSCs were generated from dental pulp stem cells using a three-step protocol. The expression of key markers, including myelin basic protein, S-100, and p75 neurotrophin receptor, was analyzed. Primary trigeminal neurons were cultured, and the expression of neurofilament 200, ß-tubulin III, and microtubule-associated protein 2 was assessed. Simultaneous culture experiments were conducted to evaluate trigeminal neuron growth in the presence of SC-DPSCs. In addition, mRNA sequencing was performed to identify key genes involved in the differentiation process, highlighting prostaglandin-endoperoxide synthase 2 (PTGS2) as a potential candidate. The results demonstrated that SC-DPSCs expressed characteristic SCs markers and facilitated axonal growth in rat trigeminal nerves. Differentiated SC-DPSCs secreted elevated levels of nerve growth factors, including brain-derived neurotrophic factor and neurotrophin-3, promoting the growth of trigeminal nerve axons. These findings suggest the regenerative potential of SC-DPSCs in dentin-dental pulp complex; PTGS2 is considered a crucial gene in this differentiation process.

Lin28 affects the proliferation and osteogenic differentiation of human dental pulp stem cells by directly inhibiting let-7b maturation.

OBJECTIVE: Activation of Lin28 gene under certain conditions promotes tissue damage repair. However, it remains unknown whether conditional expression of Lin28 facilitates the recovery of damaged pulp tissue. In the study, we focus on exploring the effects and possible regulatory mechanisms of Lin28 on the proliferation and differentiation of human dental pulp stem cells (hDPSCs). MATERIALS AND METHODS: We adopted techniques such as the ethynyl-2'-deoxyuridine (EdU) incorporation assay, RNA-protein immunoprecipitation (RIP) analysis, and luciferase assays to study the regulation of hDPSCs by Lin28. Furthermore, gain-of-function and loss-of-function analyses were also used in explored factors regulating hDPSCs activation. RESULTS: The results show that Lin28 inhibited osteogenic differentiation by directly targets pre-let-7b. Through bioinformatics sequencing and dual luciferase experiments we learned that let-7b directly targets the IGF2BP2 3'UTR. Silencing of IGF2BP2 showed a similar biological effect as overexpression of let-7b. Overexpression of IGF2BP2 counteracted the differentiation-promoting effects produced by let-7b overexpression. DISCUSSION/CONCLUSIONS: In conclusion, the RNA-binding protein Lin28 regulates osteogenic differentiation of hDPSCs by inhibiting let-7 miRNA maturation. And mature let-7b directly regulated the expression of IGF2BP2 by targeting the 3'UTR region of IGF2BP2 mRNA thus further inhibiting the differentiation of hDPSCs.

Scientific mapping of hotspots and trends of post and core research based on the Web of Science: A bibliometric analysis.

Statement of problem: Post and core (PC) restoration has been widely used to restore endodontically treated teeth. Nevertheless, bibliometric studies focusing on PC research are lacking. Purpose: The purpose of this bibliometric analysis was to review the general situation and determine the research direction of PC restoration by means of visualization, including the time development, countries, institutions, authors, journals, research categories, and the information of references with the strongest citation burst. Material and methods: The search was carried out within the topic field of the Web of Science (WoS) database. CiteSpace, VOSviewer, and R language were used to analyze the literature. Bibliometric indicators in terms of title, keyword, reference, publication time, institution, country and citation information were analyzed. Results: Between 1966 and 2022, dental research on PC trended upward, especially in 2011 and up to 2022. Scholars in Brazil, the United States, and Italy actively participated in PC research. The country that published the most was Brazil. Italy both ranked first in the H-index and average citations per item and had cooperative relations with several countries. The United States ranked first in the world in total citation count. The University of Siena published the most articles. The Journal of Prosthetic Dentistry (JPD) was the major contributing journal. The top three authors in this field were Ferrari M, Goracci C and Naumann M. The largest cooperative network of authors consisted of four Italian scholars. There were six main clusters of research topics on PC through CiteSpace co-citations. In addition, CiteSpace co-citations, CiteSpace Burst detection and VOSviewer keyword analysis showed the most important research interests, such as bond strength, fiber post, fracture resistance, and finite element analysis. Photodynamic therapy is the latest research hotspot. Finally, there were the top 25 references with the strongest citation bursts, which mainly focused on four aspects, namely, studies related to fiber posts, PC bonding strength, finite element analysis, and residual teeth and the ferrule effect. Conclusions: This bibliometric analysis provides a comprehensive overview of PC research, including the research time, country, institution, author, journal, article clustering, article keywords, and important literature analysis, so that researchers can have a clear understanding of the research situation in this field.

Effects of overtreatment with different attachment positions on maxillary anchorage enhancement with clear aligners: a finite element analysis study.

BACKGROUND: The effect of attachment positions on anchorage has not been fully explored. The aim of the present study is to analyze the effect of overtreatment with different anchorage positions on maxillary anchorage enhancement with clear aligners in extraction cases. METHODS: Models of the maxilla and maxillary dentition were constructed and imported into SOLIDWORKS software to create periodontal ligament (PDL), clear aligners, and attachments. Attachment positions on second premolars included: without attachment (WOA), buccal attachment (BA), and bucco-palatal attachment (BPA). Overtreatment degrees were divided into five groups (0°, 1°, 2°, 3°, 4°) and added on the second premolars. The calculation and analysis of the displacement trends and stress were performed using ANSYS software. RESULTS: Distal tipping and extrusion of the canines, and mesial tipping and intrusion of the posterior teeth occurred during retraction. A strong anchorage was achieved in cases of overtreatment of 2.8° with BA and 2.4° with BPA. Moreover, the BPA showed the best in achieving bodily control of the second premolars. When the overtreatment was performed, the canines and first molars also showed reduced tipping trends with second premolars attachments. And the stress on the PDL and the alveolar bone was significantly relieved and more evenly distributed in the BPA group. CONCLUSIONS: Overtreatment is an effective means for anchorage enhancement. However, the biomechanical effect of overtreatment differs across attachment positions. The BPA design performs at its best for stronger overtreatment effects with fewer adverse effects.

Sema3A Modified PDLSCs Exhibited Enhanced Osteogenic Capabilities and Stimulated Differentiation of Pre-Osteoblasts.

Genetically engineered stem cells, not only acting as vector delivering growth factors or cytokines but also exhibiting improved cell properties, are promising cells for periodontal tissue regeneration. Sema3A is a power secretory osteoprotective factor. In this study, we aimed to construct Sema3A modified periodontal ligament stem cells (PDLSCs) and evaluated their osteogenic capability and crosstalk with pre-osteoblasts MC3T3-E1. First, Sema3A modified PDLSCs was constructed using lentivirus infection system carrying Sema3A gene and the transduction efficiency was analyzed. The osteogenic differentiation and proliferation of Sema3A-PDLSCs was evaluated. Then, MC3T3-E1 was directly co-cultured with Sema3A-PDLSCs or cultured in condition medium of Sema3A-PDLSCs and the osteogenic ability of MC3T3-E1 was assessed. The results showed that Sema3A-PDLSCs expressed and secreted upregulated Sema3A protein, which confirmed successful construction of Sema3A modified PDLSCs. After osteogenic induction, Sema3A-PDLSCs expressed upregulated ALP, OCN, RUNX2, and SP7 mRNA, expressed higher ALP activity, and produced more mineralization nodes, compared with Vector-PDLSCs. Whereas, there was no obvious differences in proliferation between Sema3A-PDLSCs and Vector-PDLSCs. MC3T3-E1 expressed upregulated mRNA of ALP, OCN, RUNX2, and SP7 when directly co-cultured with Sema3A-PDLSCs than Vector-PDLSCs. MC3T3-E1 also expressed upregulated osteogenic markers, showed higher ALP activity, and produced more mineralization nodes when cultured using condition medium of Sema3A-PDLSCs instead of Vector-PDLSCs. In conclusion, our results indicated that Sema3A modified PDLSCs showed enhanced osteogenic capability, and also facilitated differentiation of pre-osteoblasts.

Bioprinted PDLSCs with high-concentration GelMA hydrogels exhibit enhanced osteogenic differentiation in vitro and promote bone regeneration in vivo.

OBJECTIVES: We aimed to explore the osteogenic potential of periodontal ligament stem cells (PDLSCs) in bioprinted methacrylate gelatine (GelMA) hydrogels in vitro and in vivo. MATERIALS AND METHODS: PDLSCs in GelMA hydrogels at various concentrations (3%, 5%, and 10%) were bioprinted. The mechanical properties (stiffness, nanostructure, swelling, and degradation properties) of bioprinted constructs and the biological properties (cell viability, proliferation, spreading, osteogenic differentiation, and cell survival in vivo) of PDLSCs in bioprinted constructs were evaluated. Then, the effect of bioprinted constructs on bone regeneration was investigated using a mouse cranial defect model. RESULTS: Ten percent GelMA printed constructs had a higher compression modulus, smaller porosity, lower swelling rate, and lower degradation rate than 3% GelMA. PDLSCs in bioprinted 10% GelMA bioprinted constructs showed lower cell viability, less cell spreading, upregulated osteogenic differentiation in vitro, and lower cell survival in vivo. Moreover, upregulated expression of ephrinB2 and EphB4 protein and their phosphorylated forms were found in PDLSCs in 10% GelMA bioprinted constructs, and inhibition of eprhinB2/EphB4 signalling reversed the enhanced osteogenic differentiation of PDLSCs in 10% GelMA. The in vivo experiment showed that 10% GelMA bioprinted constructs with PDLSCs contributed to more new bone formation than 10% GelMA constructs without PDLSCs and constructs with lower GelMA concentrations. CONCLUSIONS: Bioprinted PDLSCs with high-concentrated GelMA hydrogels exhibited enhanced osteogenic differentiation partially through upregulated ephrinB2/EphB4 signalling in vitro and promoted bone regeneration in vivo, which might be more appropriate for future bone regeneration applications. CLINICAL RELEVANCE: Bone defects are a common clinical oral problem. Our results provide a promising strategy for bone regeneration through bioprinting PDLSCs in GelMA hydrogels.

[Research progress of matrix stiffness in regulating endothelial cell sprouting].

Objective: To review the research progress on the role and mechanism of matrix stiffness in regulating endothelial cell sprouting. Methods: The related literature at home and abroad in recent years was extensively reviewed, and the behaviors of matrix stiffness related endothelial cell sprouting in different cell cultivation conditions were analyzed, and the specific molecular mechanism of matrix stiffness regulating related signal pathways in endothelial cell sprouting was elaborated. Results: In two-dimensional cell cultivation condition, increase of matrix stiffness stimulates endothelial cell sprouting within a certain range. However, in three-dimensional cell cultivation condition, the detailed function of matrix stiffness in regulating endothelial cell sprouting and angiogenesis are still unclear. At present, the research of the related molecular mechanism mainly focuses on YAP/TAZ, and roles of its upstream and downstream signal molecules. Matrix stiffness can regulate endothelial cell sprouting by activating or inhibiting signal pathways to participate in vascularization. Conclusion: Matrix stiffness plays a vital role in regulating endothelial cell sprouting, but its specific role and molecular mechanism in different environments remain ambiguous and need further study.

Bioprinting EphrinB2-Modified Dental Pulp Stem Cells with Enhanced Osteogenic Capacity for Alveolar Bone Engineering.

Bioprinting, a technology that allows depositing living cells and biomaterials together into a complex tissue architecture with desired pattern, becomes a revolutionary technology for fabrication of engineered constructs. Previously, we have demonstrated that EphrinB2-modified dental pulp stem cells (DPSCs) are expected to be promising seed cells with enhanced osteogenic differentiation capability for alveolar bone regeneration. In this study, we aimed to bioprint EphrinB2-overexpressing DPSCs with low-concentrated Gelatin methacrylate (GelMA) hydrogels into three-dimensional (3D) constructs. The printability of GelMA (5% w/v) and the structural fidelity of bioprinted constructs were examined. Then, viability, proliferation, morphology, and osteogenic differentiation of DPSCs in bioprinted constructs were measured. Finally, the effect of EphrinB2 overexpression on osteogenic differentiation of DPSCs in bioprinted constructs was evaluated. Our results demonstrated that GelMA (5% w/v) in a physical gel form was successfully bioprinted into constructs with various shapes and patterns using optimized printing parameters. Embedded DPSCs showed round-like morphology, and had a high viability (91.93% ± 8.38%) and obvious proliferation (∼1.9-fold increase) 1 day after printing. They also showed excellent osteogenic potential in bioprinted constructs. In bioprinted 3D constructs, EphrinB2-overexpressing DPSCs expressed upregulated osteogenic markers, including ALP, BMP2, RUNX2, and SP7, and generated more mineralized nodules, as compared with Vector-DPSCs. Taken together, this study indicated that fabrication of bioprinted EphrinB2-DPSCs-laden constructs with enhanced osteogenic potential was possible, and 3D bioprinting strategy combined with EphrinB2 gene modification was a promising way to create bioengineered constructs for alveolar bone regeneration.

Efficacy of antibacterial agents combined with erbium laser and photodynamic therapy in reducing titanium biofilm vitality: an in vitro study.

BACKGROUND AND OBJECTIVE: The emergence of peri-implant diseases has prompted various methods for decontaminating the implant surface. This study compared the effectiveness of three different approaches, chlorhexidine digluconate (CHX) combined with erbium-doped yttrium-aluminum-garnet (Er:YAG) laser, photodynamic therapy (PDT), and CHX only, for reducing biofilm vitality from implant-like titanium surfaces. STUDY DESIGN/MATERIALS AND METHODS: The study involved eight volunteers, each receiving a custom mouth device containing eight titanium discs. The volunteers were requested to wear the device for 72 h for biofilm development. Fluorescence microscopy was used to evaluate the remaining biofilm with a two-component nucleic acid dye kit. The vital residual biofilm was quantified as a percentage of the surface area using image analysis software. Sixty-four titanium discs were assigned randomly to one of four treatment groups. RESULTS: The percentage of titanium disc area covered by vital residual biofilm was 43.9% (7.7%), 32.2% (7.0%), 56.6% (3.6%), and 73.2% (7.8%) in the PDT, Er:YAG, CHX, and control groups, respectively (mean (SD)). Compared to the control group, the treatment groups showed significant differences in the area covered by residual biofilm (P < 0.001). CHX combined with Er:YAG laser treatment was superior to CHX combined with PDT, and CHX only was better than the control. CONCLUSION: Within the current in vitro model's limitations, CHX combined with Er:YAG laser treatment is a valid method to reduce biofilm vitality on titanium discs.

Breakthrough of extracellular vesicles in pathogenesis, diagnosis and treatment of osteoarthritis.

Osteoarthritis (OA) is a highly prevalent whole-joint disease that causes disability and pain and affects a patient's quality of life. However, currently, there is a lack of effective early diagnosis and treatment. Although stem cells can promote cartilage repair and treat OA, problems such as immune rejection and tumorigenicity persist. Extracellular vesicles (EVs) can transmit genetic information from donor cells and mediate intercellular communication, which is considered a functional paracrine factor of stem cells. Increasing evidences suggest that EVs may play an essential and complex role in the pathogenesis, diagnosis, and treatment of OA. Here, we introduced the role of EVs in OA progression by influencing inflammation, metabolism, and aging. Next, we discussed EVs from the blood, synovial fluid, and joint-related cells for diagnosis. Moreover, we outlined the potential of modified and unmodified EVs and their combination with biomaterials for OA therapy. Finally, we discuss the deficiencies and put forward the prospects and challenges related to the application of EVs in the field of OA.

[Effects of let-7a on proliferation, osteogenic differentiation and apoptosis of human dental pulp stem cells].

PURPOSE: To study the effect and possible mechanism of let-7a on proliferation, differentiation and apoptosis of human dental pulp stem cell (hDPSCs). METHODS: The cells were divided into four groups: overexpression control (let-7a control/let-7a agomir control), overexpression let-7a (let-7a mimics/let-7a agomir), knockdown let-7a control (let-7a inhibitor control) and knockdown let-7a (let-7a inhibitor). Cell counting kit-8 assay(CCK-8) was used to detect the proliferation of cells at 24 hours, 48 hours and 72 hours after transfection. Calcified nodules were detected by Alizarin red staining. The protein expression of alkaline phosphatase (ALP), osteopontin (OPN), 4E-binding protein 1 (4EBP1), p-4EBP1, mammalian target of rapamycin (mTOR) and p-mTOR were detected by Western blot. Annexin V-APC/7-AAD cell apoptosis detection kit was used to detect the level of apoptosis after transfection. Statistical analysis was performed using GraphPad Prism 5.0 software. RESULTS: Let-7a inhibited proliferation of hDPSCs and promoted odontoblast differentiation and apoptosis. Let-7a down-regulated the expression of 4EBP1, p-4EBP1, mTOR and p-mTOR. CONCLUSIONS: Let-7a may inhibit proliferation of hDPSCs and promote their differentiation and apoptosis by inhibiting mTOR-4EBP1 molecular pathway.

Effectiveness of the attachment position in molar intrusion with clear aligners: a finite element study.

OBJECTIVE: To evaluate the biomechanical effects of different attachments' position for maxillary molar intrusion with clear aligner treatment by finite element analysis. METHODS: Cone-beam computed tomography images of a patient with supra-eruption of the maxillary second molars were selected to construct three-dimensional models of the maxilla, periodontal ligaments, dentition, and clear aligner. The models were divided into four groups depending on the attachment location on the first molar: (1) no attachment (NA), (2) buccal attachment (BA), (3) palatal attachment (PA), and (4) bucco-palatal attachment (BPA). After applying an intrusion of 0.2 mm on the second molar, displacements and stress distributions of the teeth, aligner, and periodontal ligament were analyzed with the finite element software. RESULTS: All groups displayed equivalent movement patterns of aligners. The NA and BA groups showed buccal tipping of the second molar, while the PA group showed palatal tipping. The BPA group had the highest intruding value and the lowest buccal/palatal tipping value. All groups showed mesial tipping of the second molar. Stress distribution in the periodontal ligament strongly correlated with the attachment position. The BPA group showed the best stress distribution. CONCLUSION: Combined BA and PA could effectively prevent buccal and palatal tipping and showed the best efficiency in intruding the second molar. The second molar showed an unavoidable tendency to tip mesially, regardless of the attachment position.

Clinical applications of concentrated growth factors membrane for sealing the socket in alveolar ridge preservation: a randomized controlled trial.

The purpose of this study was to evaluate the efficacy of concentrated growth factor (CGF) membrane for the sealing of alveolar socket in alveolar ridge preservation (ARP). A total of 22 patients with 24 alveolar sockets were recruited and divided randomly into CGF group and Bio-Gide collagen membrane group. The soft tissue wound healing rate was calculated using intraoral scanner at 3, 7, and 14 days after ARP, and the bone resorption volume at 1, 3, and 5 mm below the alveolar ridge was measured by CBCT at 6 months postoperation. The keratinized gingival width was also measured before and 6 months after ridge preservation. In terms of soft tissue healing rate, the CGF group exhibited significant higher than that of Bio-Gide group at both 7 and 14 days after surgery (P < 0.05). However, there was no significantly different in bone resorption rate and the width of keratinized gingival after 6 months (P > 0.05). Therefore, the use of CGFs membranes for wound closure in ARP is a reliable method, but more clinical data are needed to prove it.

Strontium-doping promotes bone bonding of titanium implants in osteoporotic microenvironment.

Osteoporosis is a major challenge to oral implants, and this study focused on improving the osseointegration ability of titanium (Ti) implants in osteoporosis environment via surface modification, including doping of strontium ion and preparation of nanoscale surface feature. Our previous studies have shown that strontium (Sr) ions can enhance osteogenic activity. Therefore, we aimed to comprehensively evaluate the effect of hydrothermal treatment of Sr-doped titanium implant coating on bone-binding properties in the microenvironment of osteoporosis in this study. We fabricated Sr-doped nanocoating (AHT-Sr) onto the surface of titanium implants via hydrothermal reaction. The rough Sr-doping had good biological functions and could apparently promote osteogenic differentiation of osteoporotic bone marrow mesenchymal stem cells (OVX-BMSCs). Most importantly, AHT-Sr significantly promoted bone integration in the osteoporosis environment. This study provides an effective approach to implant surface modification for better osseointegration in an osteoporotic environment.

The potential therapeutic role of extracellular vesicles in osteoarthritis.

Osteoarthritis (OA) is a worldwide and disabling disease, which cause severe pain and heavy socioeconomic burden. However, pharmacologic or surgical therapies cannot mitigate OA progression. Mesenchymal stem cells (MSCs) therapy has emerged as potential approach for OA treatment, while the immunogenicity and ethical audit of cell therapy are unavoidable. Compared with stem cell strategy, EVs induce less immunological rejection, and they are more stable for storage and in vivo application. MSC-EVs-based therapy possesses great potential in regulating inflammation and promoting cartilage matrix reconstruction in OA treatment. To enhance the therapeutic effect, delivery efficiency, tissue specificity and safety, EVs can be engineered via different modification strategies. Here, the application of MSC-EVs in OA treatment and the potential underlying mechanism were summarized. Moreover, EV modification strategies including indirect MSC modification and direct EV modification were reviewed.

Modifying a 3D-Printed Ti6Al4V Implant with Polydopamine Coating to Improve BMSCs Growth, Osteogenic Differentiation, and In Situ Osseointegration In Vivo.

Nowadays, 3D printing technology has been applied in dentistry to fabricate customized implants. However, the biological performance is unsatisfactory. Polydopamine (PDA) has been used to immobilize bioactive agents on implant surfaces to endow them with multiple properties, such as anti-infection and pro-osteogenesis, benefiting rapid osseointegration. Herein, we fabricated a PDA coating on a 3D-printed implant surface (3D-PDA) via the in situ polymerization method. Then the 3D-PDA implants' pro-osteogenesis capacity and the osseointegration performance were evaluated in comparison with the 3D group. The in vitro results revealed that the PDA coating modification increased the hydrophilicity of the implants, promoting the improvement of the adhesion, propagation, and osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) in vitro. Additionally, the 3D-PDA implant improved osteointegration performance in vivo. The present study suggested that PDA coating might be a feasible strategy to optimize 3D-printed implant surfaces, making a preliminary research basis for the subsequent work to immobilize bioactive factors on the 3D-printed implant surface.

Small molecules efficiently reprogram apical papilla stem cells into neuron-like cells.

Stem cell-based therapy may provide a novel approach for neural tissue regeneration. A small molecule cocktail-based culture protocol was previously shown to enhance neurogenic differentiation of stem cells from dental tissues. The present study aimed to investigate the early phase of small molecule-induced neurogenic differentiation of stem cells from the apical papilla (SCAP). SCAP were cultured in neural-induction medium or neural-induction medium with small molecules (NIMS-SCAP) and examined for their cell morphologies. Expression levels of neural progenitor cell-related markers, including Nestin, paired-box gene 6 (Pax6) and Sry-related HMG box 2 (Sox2), were examined using western blotting and immunocytofluorescence. Expression of differentiated neuron-related markers, including neurofilament protein (NFM), neuron-specific nuclear protein (NeuN) and microtubule-associated protein (MAP)-2, were also examined using western blotting, while NFM and MAP2 gene expression and cell proliferation were assessed using reverse transcription-quantitative (RT-q)PCR and Cell Counting Kit (CCK)-8 assays, respectively. SCAP morphology was affected by small molecules after as little as 30 min. Specifically, Nestin, Pax6 and Sox2 expression detected using western blotting was increased by day 3 but then decreased over the course of 7 days with neural induction, while immunocytofluorescence revealed expression of all three markers in NIMS-SCAP. The protein levels of NFM, NeuN and MAP2 on day 7 were significantly upregulated in NIMS-SCAP, as detected using western blotting, while NFM and MAP2 gene expression levels detected using RT-qPCR were significantly increased on days 5 and 7. Proliferation of NIMS-SCAP ceased after 5 days. Electrophysiological analysis showed that only SCAP cultured in NIMS had the functional activity of neuronal cells. Thus, small molecules reprogrammed SCAP into neural progenitor cells within the first 3 days, followed by further differentiation into neuron-like cells.