DNA Tetrahedron Delivering miR-21-5p Promotes Senescent Bone Defects Repair through Synergistic Regulation of Osteogenesis and Angiogenesis.

Compromised osteogenesis and angiogenesis is the character of stem cell senescence, which brought difficulties for bone defects repairing in senescent microenvironment. As the most abundant bone-related miRNA, miRNA-21-5p plays a crucial role in inducing osteogenic and angiogenic differentiation. However, highly efficient miR-21-5p delivery still confronts challenges including poor cellular uptake and easy degradation. Herein, TDN-miR-21-5p nanocomplex is constructed based on DNA tetrahedral (TDN) and has great potential in promoting osteogenesis and alleviating senescence of senescent bone marrow stem cells (O-BMSCs), simultaneously enhancing angiogenic capacity of senescent endothelial progenitor cells (O-EPCs). Of note, the activation of AKT and Erk signaling pathway may direct regulatory mechanism of TDN-miR-21-5p mediated osteogenesis and senescence of O-BMSCs. Also, TDN-miR-21-5p can indirectly mediate osteogenesis and senescence of O-BMSCs through pro-angiogenic growth factors secreted from O-EPCs. In addition, gelatin methacryloyl (GelMA) hydrogels are mixed with TDN and TDN-miR-21-5p to fabricate delivery scaffolds. TDN-miR-21-5p@GelMA scaffold exhibits greater bone repair with increased expression of osteogenic- and angiogenic-related markers in senescent critical-size cranial defects in vivo. Collectively, TDN-miR-21-5p can alleviate senescence and induce osteogenesis and angiogenesis in senescent microenvironment, which provides a novel candidate strategy for senescent bone repair and widen clinical application of TDNs-based gene therapy.

ScRNA-seq links dental fibroblasts heterogeneity with mechanoresponsiveness.

BACKGROUND AND OBJECTIVE: Periodontal ligament (PDL) and dental pulp (DP) share a common origin but have distinct biological and mechanical functions. To what extent the mechanoresponsive property of PDL can be attributed to its unique transcriptional profiles of cellular heterogeneity is unclear. This study aims to decipher cellular heterogeneity and distinct mechanoresponsive characteristics of odontogenic soft tissues and their underlying molecular mechanisms. MATERIALS AND METHODS: A single-cell comparison of digested human periodontal ligament (PDL) and dental pulp (DP) was performed using scRNA-seq. An in vitro loading model was constructed to measure mechanoresponsive ability. Dual-luciferase assay, overexpression, and shRNA knockdown were used to investigate the molecular mechanism. RESULTS: Our results demonstrate striking fibroblast heterogeneity across and within human PDL and DP. We demonstrated that a tissue-specific subset of fibroblasts existed in PDL exhibiting high expression of mechanoresponsive extracellular matrix (ECM) genes, which was verified by an in vitro loading model. ScRNA-seq analysis indicated a particularly enriched regulator in PDL-specific fibroblast subtype, Jun Dimerization Protein 2 (JDP2). Overexpression and knockdown of JDP2 extensively regulated the downstream mechanoresponsive ECM genes in human PDL cells. The force loading model demonstrated that JDP2 responded to tension and that knockdown of JDP2 effectively inhibited the mechanical force-induced ECM remodeling. CONCLUSIONS: Our study constructed the PDL and DP ScRNA-seq atlas to demonstrate PDL and DP fibroblast cellular heterogeneity and identify a PDL-specific mechanoresponsive fibroblast subtype and its underlying mechanism.

Mandibular stability and condylar changes following orthognathic surgery in mandibular hypoplasia patients associated with preoperative condylar resorption.

OBJECTIVES: To evaluate postoperative mandibular stability and condylar changes in patients with mandibular hypoplasia and preoperative condylar resorption (CR) undergoing orthognathic surgery. MATERIALS AND METHODS: Fifty-four patients were included in this retrospective study. Computed tomography (CT) scans were acquired preoperatively (T0), 2-7 days immediate postoperatively (T1), and at least 1 year postoperatively (T2). Three-dimensional (3D) cephalometric analysis and measurements of condylar angle, volume, and position (joint spaces) were performed. A 2-mm mandibular relapse was deemed clinically acceptable. We also analyzed the correlations between relapse and postoperative CR and susceptible factors using a multivariate logistic regression model. RESULTS: The results showed one year after the surgery, the average mandibular relapse was 1.0 mm (p < 0.05), and the average reduction of condylar volume was 152.4 mm3 (12.7%). Condyle-fossa relationships were improved immediately after the surgery, with a tendency of returning to their original state in the follow-up (p < 0.05). Anteroposterior advancement at point B (B-CP advancement) at T1 and superior joint space (SJS) at T0 were significantly correlated with mandibular relapse, and postoperative CR was mainly associated with vertical increasement at point B (B-AP increasement) at T1. The optimal cut-off values were as follows: 1.6 mm for SJS, 4.2 mm for B-CP advancement, and 1.8 mm for B-AP increasement. Concomitant advancement Genioplasty showed no significant correlation with relapse and postoperative CR. CONCLUSIONS: While patients with mandibular hypoplasia and preoperative CR were vulnerable to further condylar resorption after mandibular advancement, the treatment outcomes were generally clinically acceptable. Postoperative relapse was associated with a larger than 4.2 mm of mandibular advancement measured at B-CP and a larger than 1.6 mm of superior joint space measured at SJS, and postoperative CR was associated with a larger than 1.8 mm of mandibular vertical increasement measured at B-AP. CLINICAL RELEVANCE: The findings of this study suggested that the mandibular advancement might be limited to 5 mm for patients with preoperative CR. A concomitant advancement genioplasty might also be considered to achieve a better facial profile in these patients.

Reducing relapse and accelerating osteogenesis in rapid maxillary expansion using an injectable mesoporous bioactive glass/fibrin glue composite hydrogel.

Rapid maxillary expansion (RME), as a common treatment for craniomaxillofacial deformity, faces the challenge of high relapse rates and unsatisfactory therapeutic effects. In this study, a standardized Sprague-Dawley (SD) rat RME model was first established with a modified expander as well as retainer design and optimized anterior maxillary expanding force of 100 g which exerted the most synchronized mobility of mid-palatal suture and incisors. Via the standardized model, the high relapse rate was proven to be attributed to insufficient osteogenesis in expanded suture, requiring long-term retainer wearing in clinical situations. To reduce the relapse rate, mesoporous bioactive glass/fibrin glue (MBG/FG) composite hydrogels were developed for an in situ minimal invasive injection that enhance osteogenesis in the expanded palate. The component of 1 wt% MBG was adopted for enhanced mechanical strength, matched degradation rate and ion dissolution, excellent in vitro biocompatibility and osteoinductivity. Effects of 1%MBG/FG composite hydrogel on osteogenesis in expanded mid-palatal sutures with/without retention were evaluated in the standardized model. The results demonstrated that injection of 1%MBG/FG composite hydrogel significantly promoted bone formation within the expanded mid-palatal suture, inhibited osteoclastogenesis and benefited the balance of bone remodeling towards osteogenesis. Combination of retainer and injectable biomaterial was demonstrated as a promising treatment to reduce relapse rate and enhance osteogenesis after RME. The model establishment and the composite hydrogel development in this article might provide new insight to other craniomaxillofacial deformity treatment and design of bone-repairing biomaterials with higher regenerative efficiency.

Effect of Swell1 on regulating chondrocyte hypertrophy during the condylar osteochondral development process in mice.

Chondrocyte hypertrophy is a significant factor in cartilage development, yet the molecular mechanism for cell volume expand during the process is remains unclear. In the present study, the relationship between Swell1, a cell volume regulated anion channel, and chondrocyte hypertrophy was explored. The results reveal that the spatiotemporal expression of Swell1 was similar with the development process of hypertrophic chondrocytes in condyles. Through Col10a1 mediated knock out of Swell1 in hypertrophy chondrocytes, we found that there are less obvious boundary between different condylar cartilage layers in which increased hypertrophic chondrocytes were scattered in all three cartilage layers. The cortical bone mass and bone mineral density in the subchondral bone significantly increased. Additionally, knock out of Swell1 could increase the expression of OCN in the femur condyle. Based on the aforementioned findings, a conclusion could be drawn that Swell1 is a significant factor in chondrocyte hypertrophy during the condylar osteochondral development process, and there was some difference between the mandibular and femur condyles, which will provide some new clues for understanding the development of cartilage and related diseases.

Optimized BMSC-derived osteoinductive exosomes immobilized in hierarchical scaffold via lyophilization for bone repair through Bmpr2/Acvr2b competitive receptor-activated Smad pathway.

Mesenchymal stem cell-derived exosomes (MSC-exos), with its inherent capacity to modulate cellular behavior, are emerging as a novel cell-free therapy for bone regeneration. Herein, focusing on practical applying problems, the osteoinductivity of MSC-exos produced by different stem cell sources (rBMSCs/rASCs) and culture conditions (osteoinductive/common) were systematically compared to screen out an optimized osteogenic exosome (BMSC-OI-exo). Via bioinformatic analyses by miRNA microarray and in vitro pathway verification by gene silencing and miRNA transfection, we first revealed that the osteoinductivity of BMSC-OI-exo was attributed to multi-component exosomal miRNAs (let-7a-5p, let-7c-5p, miR-328a-5p and miR-31a-5p). These miRNAs targeted Acvr2b/Acvr1 and regulated the competitive balance of Bmpr2/Acvr2b toward Bmpr-elicited Smad1/5/9 phosphorylation. On these bases, lyophilized delivery of BMSC-OI-exo on hierarchical mesoporous bioactive glass (MBG) scaffold was developed to realize bioactivity maintenance and sustained release by entrapment in the surface microporosity of the scaffold. In a rat cranial defect model, the loading of BMSC-OI-exo efficiently enhanced the bone forming capacity of the scaffold and induced rapid initiation of bone regeneration. This paper could provide empirical bases of MSC-exo-based therapy for bone regeneration and theoretical bases of MSC-exo-induced osteogenesis mechanism. The BMSC-OI-exo-loaded MBG scaffold developed here represented a promising bone repairing strategy for future clinical application.

An overview of polyester/hydroxyapatite composites for bone tissue repairing.

OBJECTIVES: The polyester/hydroxyapatite (polyester/HA) composites play an important role in bone tissue repairing, mostly because they mimic the composition and structure of naturally mineralized bone tissue. This review aimed to discuss commonly used geometries of polyester/HA composites, including microspheres, membranes, scaffolds and bulks, and their applications in bone tissue repairing and to discuss existed restrictions and developing trends of polyester/HA. METHODS: The current review was conducted by searching Web of Science, and Google Scholar for relevant studies published related with polyester/HA composites. Selected studies were analyzed with a focus on the fabrication techniques, properties (mechanical properties, biodegradable properties and biological properties) and applications of polyester/HA composites in bone repairing. RESULTS: A total of 111 articles were introduced to discuss the review. Different geometries of polyester/HA composites were discussed. In addition, properties and applications of polyester/HA composites were evaluated. The addition of HA into polyester can adjust the mechanical and biodegradability of composites. Besides, the addition of HA into polyester can improve its osteogenic abilities. The results showed that polyester/HA composites can ideal candidate for bone tissue repairing. CONCLUSION: Polyester/HA composites have many remarkable properties, such as appropriate mechanical strength, biodegradability, favorable biological properties. Diverse geometries of polyester/HA composites have been used in bone repairing, drug delivery and implant fixation. Further work needs to be done to investigate existed restrictions, including the controlled degradation rate, controlled drug release performance, well-matched mechanical properties, and novel fabrication techniques. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: The present review reveals the current state of the polyester/HA composites used in bone tissue repairing, contributing to future trends of polyester/HA composites in the forthcoming future.

Experimental and clinical evaluation of BMP2-CPC graft versus deproteinized bovine bone graft for guided bone regeneration: A pilot study.

In this study, we proposed BMP2-incorporated calcium phosphate cement (BMP2-CPC), for application in guided bone regeneration (GBR) and compared the experimental bone restoration performance and clinical alveolar bone reconstruction outcome of BMP2-CPC with those of deproteinized bovine bone (DBB). The animal study indicated that, compared to DBB, which induced the slow ingrowth of new bone, BMP2-CPC induced numerous small growth centers for bone regeneration and facilitated a significant amount of bone regeneration in rabbit calvarial bone defects. Fewer residual graft particles remained in the BMP2-CPC-treated defects than in the DBB-treated defects. The clinical study indicated that BMP2-CPC was similar to DBB in remedying alveolar bone insufficiency and maintaining implant stability. In conclusion, the results of this present study indicate that compared to DBB, BMP2-CPC can significantly enhance in vivo bone regeneration and remodeling in rabbit calvarial bone defects and shows preliminary support on its clinical application in GBR surgeries.

A viscoelastic PEGylated poly(glycerol sebacate)-based bilayer scaffold for cartilage regeneration in full-thickness osteochondral defect.

Defects of either articular cartilage or subchondral bone would destroy the structural integrity and functionality of the joint. Reconstruction of osteochondral defects requires difunctional scaffolds that simultaneously induce cartilage and subchondral bone morphogenesis, however, high-performance cartilage reconstructive scaffolds remain a considerable challenge. In this study, a solvent-free urethane crosslinking and spontaneous pore-forming procedure under room temperature was proposed and optimized to produce PEGylated poly(glycerol sebacate) (PEGS) scaffolds with controllable crosslinking degrees and hierarchical macro-/micro-porosities. Based on the economical and feasible preparative approach, the viscoelastic PEGS-12h with low crosslinking degree was demonstrated to significantly stimulate chondrogenic differentiation, maintain chondrocyte phenotype and enhance cartilage matrix secretion compared to elastic polymer with high crosslinking degree, emphasizing the importance of matrix viscoelasticity in cartilage regeneration. On this basis, the viscoelastic low-crosslinked PEGS-12h was combined with the well-acknowledged osteoinductive mesoporous bioactive glass (MBG) to construct a difunctional PEGS/MBG bilayer scaffold, and evaluated in a full-thickness osteochondral defect model in vivo. The PEGS/MBG bilayer scaffold successfully reconstructed well-integrated articular hyaline cartilage and its subchondral bone in 12 weeks, exhibiting extraordinary regenerative efficiency. The results indicated that the viscoelastic PEGS scaffold and PEGS/MBG bilayer scaffold proposed in this study made an excellent candidate for cartilage and osteochondral regeneration, and was expected for clinical translation in the future.

Perinatal stem cells: A promising cell resource for tissue engineering of craniofacial bone.

In facing the mounting clinical challenge and suboptimal techniques of craniofacial bone defects resulting from various conditions, such as congenital malformations, osteomyelitis, trauma and tumor resection, the ongoing research of regenerative medicine using stem cells and concurrent advancement in biotechnology have shifted the focus from surgical reconstruction to a novel stem cell-based tissue engineering strategy for customized and functional craniofacial bone regeneration. Given the unique ontogenetical and cell biological properties of perinatal stem cells, emerging evidence has suggested these extraembryonic tissue-derived stem cells to be a promising cell source for extensive use in regenerative medicine and tissue engineering. In this review, we summarize the current achievements and obstacles in stem cell-based craniofacial bone regeneration and subsequently we address the characteristics of various types of perinatal stem cells and their novel application in tissue engineering of craniofacial bone. We propose the promising feasibility and scope of perinatal stem cell-based craniofacial bone tissue engineering for future clinical application.