Tooth defect and tooth loss are common clinical diseases in stomatology. Compared with the traditional oral restoration treatment, tooth regeneration has unique advantages and is currently the focus of oral biomedical research. It is known that dozens of cytokines/growth factors and other bioactive factors are expressed in a spatial-temporal pattern during tooth development. On the other hand, the technology for spatial-temporal control of drug release has been intensively studied and well developed recently, making control release of these bioactive factors mimicking spatial-temporal pattern more feasible than ever for the purpose of tooth regeneration. This article reviews the research progress on the tooth development and discusses the future of tooth regeneration in the context of spatial-temporal release of developmental factors.

Mesenchymal stem cells (MSCs) are considered the most promising seed cells for regenerative medicine because of their considerable therapeutic properties and accessibility. Fine-tuning of cell biological processes, including differentiation and senescence, is essential for achievement of the expected regenerative efficacy. Researchers have recently made great advances in understanding the spatiotemporal gene expression dynamics that occur during osteogenic, adipogenic and chondrogenic differentiation of MSCs and the intrinsic and environmental factors that affect these processes. In this context, histone modifications have been intensively studied in recent years and have already been indicated to play significant and universal roles in MSC fate determination and differentiation. In this review, we summarize recent discoveries regarding the effects of histone modifications on MSC biology. Moreover, we also provide our insights and perspectives for future applications.

Rationale: Spatial-temporal control of cell fate in vivo is of great importance for regenerative medicine. Currently, there remain no practical strategies to tune cell-fate spatial-temporally. Optogenetics is a biological technique that widely used to control cell activity in genetically defined neurons in a spatiotemporal-specific manner by light. In this study, optogenetics was repurposed for precise bone tissue regeneration.Methods: Lhx8 and BMP2 genes, which are considered as the master genes for mesenchymal stem cell proliferation and differentiation respectively, were recombined into a customized optogenetic control system. In the system, Lhx8 was constitutively expressed, while BMP2 together with shLhx8 expression was driven by blue light.Results: As expected, blue light induced BMP2 expression and inactivated Lhx8 expression in cells infected with the optogenetic control system. Optogenetic control of BMP2 and Lhx8 expression inversely regulates MSC fate in vitro. By animal study, we found that blue light could fine-tune the regeneration in vivo. Blue light illumination significantly promotes bone regeneration when the scaffold was loaded with MSCs infected with adeno-Lhx8, GI-Gal4DBD, LOV-VP16, and BMP2-shLhx8.Conclusions: Together, our study revealed that optogenetic control of the master genes for mesenchymal stem cell proliferation and differentiation would be such a candidate strategy for precise regenerative medicine.

Mechanical forces play important roles in many biological processes and there is increasing interest and understanding of these roles. Mechanotransduction is the process by which mechanical stimuli are converted to biochemical signals through specific mechanisms, and this results in the activation of downstream signaling pathways with specific effects on cell behaviors. This review systematically summarizes the current understanding of the mechanosensitive Piezo1 and Piezo2 ion channels in craniofacial bone, tooth, and periodontal tissue, presenting the latest relevant evidence with implications for potential treatments and managements of dental and orofacial diseases and deformities. The mechanosensitive ion channels Piezo1 and Piezo2 are widely expressed in various cells and tissues and have essential functions in mechanosensation and mechanotransduction. These channels play an active role in many physiological and pathological processes, such as growth and development, mechano-stimulated bone homeostasis and the mediation of inflammatory responses. Emerging evidence indicates the expression of Piezo1 and Piezo2 in bone, dental tissues and dental tissue-derived stem cells and suggests that they function in dental sensation transduction, dentin mineralization and periodontal bone remodeling and modulate orthodontic tooth movement.

Background The spatiotemporal regulation of essential genes is crucial for controlling the growth and differentiation of cells in a precise manner during regeneration. Recently, optogenetics was considered as a potent technology for sophisticated regulation of target genes, which might be a promising tool for regenerative medicine. In this study, we used an optogenetic control system to precisely regulate the expression of Lhx8 to promote efficient bone regeneration. Methods Quantitative real-time PCR and western blotting were used to detect the expression of Lhx8 and osteogenic marker genes. Alkaline phosphatase staining and alizarin red staining were used to detect alkaline phosphatase activity and calcium nodules. A customized optogenetic expression system was constructed to regulate Lhx8, of which the expression was activated in blue light but not in dark. We also used a critical calvarial defect model for the analysis of bone regeneration in vivo. Moreover, micro-computed tomography (micro-CT), three-dimensional reconstruction, quantitative bone measurement, and histological and immunohistochemistry analysis were performed to investigate the formation of new bone in vivo. Results During the osteogenic differentiation of BMSCs, the expression levels of Lhx8 increased initially but then decreased thereafter. Lhx8 promoted the early proliferation of BMSCs but inhibited subsequent osteogenic differentiation. The optogenetic activation of Lhx8 in BMSCs in the early stages of differentiation by blue light stimulation led to a significant increase in cell proliferation, thus allowing a sufficient number of differentiating BMSCs to enter the later osteogenic differentiation stage. Analysis of the critical calvarial defect model revealed that the pulsed optogenetic activation of Lhx8 in transplanted BMSCs over a 5-day period led to a significant increase in the generation of bone in vivo. Conclusions Lhx8 plays a critical role in balancing proliferation and osteogenic differentiation in BMSCs. The optogenetic activation of Lhx8 expression at early stage of BMSCs differentiation led to better osteogenesis, which would be a promising strategy for precise bone regeneration.

Periodontitis is the sixth most prevalent chronic disease globally and places significant burdens on societies and economies worldwide. Behavioral modification, risk factor control, coupled with cause-related therapy have been the “gold standard” treatment for managing periodontitis. Given that host inflammatory and immunological responses play critical roles in the pathogenesis of periodontitis and impact treatment responses, several adjunctive strategies aimed at modulating host responses and improving the results of periodontal therapy and maintenance have been proposed. Of the many pharmacological host modulators, we focused on non-steroidal anti-inflammatory drugs (NSAIDs), due to their long history and extensive use in relieving inflammation and pain and reducing platelet aggregation. NSAIDs have been routinely indicated for treating rheumatic fever and osteoarthritis and utilized for the prevention of cardiovascular events. Although several efforts have been made to incorporate NSAIDs into the treatment of periodontitis, their effects on periodontal health remain poorly characterized, and concerns over the risk–benefit ratio were also raised. Moreover, there is emerging evidence highlighting the potential of NSAIDs, especially aspirin, for use in periodontal regeneration. This review summarizes and discusses the use of NSAIDs in various aspects of periodontal therapy and regeneration, demonstrating that the benefits of NSAIDs as adjuncts to conventional periodontal therapy remain controversial. More recent evidence suggests a promising role for NSAIDs in periodontal tissue engineering and regeneration.

The homeobox gene, LIM‐homeobox 8 (Lhx8), has previously been identified as an essential transcription factor for dental mesenchymal development. However, how Lhx8 itself is regulated and regulates odontogenesis remains poorly understood. In this study, we employed an RNAscope assay to detect the co‐expression pattern of Lhx8 and Suv39h1 in the dental mesenchyme, which coincided with the dynamic expression profiles of the early epithelium signal of Fibroblast Growth Factor 8 (FGF8) and the later mesenchymal signal Bone Morphogenetic Protein 2 (BMP2). Moreover, FGF8 activated Lhx8, whereas BMP2 repressed Lhx8 expression at the transcriptional level. The high expression of Lhx8 in the early dental mesenchyme maintained the cell fate in an undifferentiated status by interacting with Suv39h1, a histone‐lysine N‐methyltransferase constitutively expressed in the dental mesenchyme. Further in the ex vivo organ culture model, the knockdown of Suv39h1 significantly blocked the function of Lhx8 and FGF8. Mechanistically, Lhx8/Suv39h1 recognized the odontoblast differentiation‐related genes and repressed gene expression via methylating H3K9 on their promoters. Taken together, our data here suggest that Lhx8/Suv39h1 complex is inversely regulated by epithelium‐mesenchymal signals, balancing the differentiation and proliferation of dental mesenchyme via H3K9 methylation.

Background and objective The effect of probiotics on oral health maintenance in orthodontic patients remains controversial. The aim of the study is to systematically review and assess the effects of probiotics on the oral health and microbiome of patients undergoing orthodontic treatment. Search methods and selection criteria Databases including PubMed, Web of Science, Cochrane Library, ClinicalTrials.gov, and ProQuest Dissertations & Theses Global databases were searched from their inception until June 2022. Randomised controlled trials that assessed the effects of probiotics on clinical and microbial outcomes in patients undergoing orthodontic treatment were included. Data collection and analysis Data screening and collection were performed, and the risk of bias (RoB) was assessed using the Cochrane RoB 2 tool. The meta-analysis evaluated the effects of probiotics on Streptococcus mutans (S. mutans) and Lactobacillus counts. The quality of the evidence from the meta-analyses was assessed with Grading of Recommendations Assessment, Development and Evaluation (GRADE). Results A total of 405 records were identified, of which 15 studies were included in the qualitative synthesis and 4 in the meta-analysis. The patients in all the included studies were treated with fixed orthodontic appliances. Results regarding clinical outcomes were controversial; four out of five studies reported no significant changes in plaque in the probiotic group (P > .05), and two out of three studies reported no significant changes in the gingival index (P > .05). Regarding microbial outcomes, the meta-analysis results revealed that probiotics significantly increased the likelihood of reducing the abundance of S. mutans to below 105 CFU/ml (risk ratio: 2.05 [1.54, 2.72], P < .001) and reduced the likelihood of increasing the abundance of S. mutans to beyond 106 CFU/ml (risk ratio: 0.48 [0.28, 0.83], P = .009). However, the quality of evidence according to the GRADE was moderate. Conclusions and implications There is insufficient evidence to determine the clinical benefits of probiotics as a supplement for the oral health of patients undergoing orthodontic treatment. However, probiotics may have benefits in reducing the salivary S. mutans counts in orthodontic patients. Registration PROSPERO (CRD42022366650).