Christine Hong

A Unifying Model for the Selective Regulation of Inducible Transcription by CpG Islands and Nucleosome Remodeling

We describe a broad mechanistic framework for the transcriptional induction of mammalian primary response genes by Toll-like receptors and other stimuli. One major class of primary response genes is characterized by CpG-island promoters, which facilitate promiscuous induction from constitutively active chromatin without a requirement for SWI/SNF nucleosome remodeling complexes. The low nucleosome occupancy at promoters in this class can be attributed to the assembly of CpG islands into unstable nucleosomes, which may lead to SWI/SNF independence. Another major class consists of non-CpG-island promoters that assemble into stable nucleosomes, resulting in SWI/SNF dependence and a requirement for transcription factors that promote selective nucleosome remodeling. Some stimuli, including serum and tumor necrosis factor-alpha, exhibit a strong bias toward activation of SWI/SNF-independent CpG-island genes. In contrast, interferon-beta is strongly biased toward SWI/SNF-dependent non-CpG-island genes. By activating a diverse set of transcription factors, Toll-like receptors induce both classes and others for an optimal response to microbial pathogens.

Histone methyltransferases and demethylases: regulators in balancing osteogenic and adipogenic differentiation of mesenchymal stem cells.

Mesenchymal stem cells (MSCs) are characterized by their self-renewing capacity and differentiation potential into multiple tissues. Thus, management of the differentiation capacities of MSCs is important for MSC-based regenerative medicine, such as craniofacial bone regeneration, and in new treatments for metabolic bone diseases, such as osteoporosis. In recent years, histone modification has been a growing topic in the field of MSC lineage specification, in which the Su(var)3–9, enhancer-of-zeste, trithorax (SET) domain-containing family and the Jumonji C (JmjC) domain-containing family represent the major histone lysine methyltransferases (KMTs) and histone lysine demethylases (KDMs), respectively. In this review, we summarize the current understanding of the epigenetic mechanisms by which SET domain-containing KMTs and JmjC domain-containing KDMs balance the osteogenic and adipogenic differentiation of MSCs.

Distalization of the entire maxillary arch in an adult

Many appliances are available to move maxillary molars distally. First molars have routinely been moved distally with nickel-titanium coil springs and nickel-titanium wire before the eruption of the second molars. However, when first molars are moved distally after the eruption of the second molars, they tend to move slowly, and anchorage loss increases. In adults, the midpalatal area is appropriate for placing titanium miniscrews for orthodontic anchorage. This case report demonstrates the ability of midpalatal miniscrews to control anchorage while distalizing the entire maxillary dentition in an adult, with improvements in lip profile resulting from the retraction of anterior teeth followed by a good response of the lips. This report suggests that absolute anchorage can be established by placing miniscrews in the palate and that miniscrew anchorage can serve as anchorage for the distal movement of an entire arch.

KDM6B epigenetically regulates odontogenic differentiation of dental mesenchymal stem cells

Mesenchymal stem cells (MSCs) have been identified and isolated from dental tissues, including stem cells from apical papilla, which demonstrated the ability to differentiate into dentin-forming odontoblasts. The histone demethylase KDM6B (also known as JMJD3) was shown to play a key role in promoting osteogenic commitment by removing epigenetic marks H3K27me3 from the promoters of osteogenic genes. Whether KDM6B is involved in odontogenic differentiation of dental MSCs, however, is not known. Here, we explored the role of KDM6B in dental MSC fate determination into the odontogenic lineage. Using shRNA-expressing lentivirus, we performed KDM6B knockdown in dental MSCs and observed that KDM6B depletion leads to a significant reduction in alkaline phosphate (ALP) activity and in formation of mineralized nodules assessed by Alizarin Red staining. Additionally, mRNA expression of odontogenic marker gene SP7 (osterix, OSX), as well as extracellular matrix genes BGLAP (osteoclacin, OCN) and SPP1 (osteopontin, OPN), was suppressed by KDM6B depletion. When KDM6B was overexpressed in KDM6B-knockdown MSCs, odontogenic differentiation was restored, further confirming the facilitating role of KDM6B in odontogenic commitment. Mechanistically, KDM6B was recruited to bone morphogenic protein 2 (BMP2) promoters and the subsequent removal of silencing H3K27me3 marks led to the activation of this odontogenic master transcription gene. Taken together, our results demonstrated the critical role of a histone demethylase in the epigenetic regulation of odontogenic differentiation of dental MSCs. KDM6B may present as a potential therapeutic target in the regeneration of tooth structures and the repair of craniofacial defects.

Stability comparison between commercially available mini-implants and a novel design: part 1.

OBJECTIVE To compare mechanical stability among five mini-implant designs--a newly invented design and four commercially available designs that vary by shape and threading; to calculate external surface area of each design using high-resolution micro-computed tomography; and to evaluate the relationship between surface area and stability results. MATERIALS AND METHODS The four commercially available mini-implants--single-threaded and cylindrical (SC), single-threaded and tapered (ST), double-threaded and cylindrical (DC), double-threaded and tapered (DT)--and a new implant that is designed to engage mostly in cortical bone with shorter and wider dimensions (N1) were inserted in simulated bone with cortical and trabecular bone layers. The mechanical study consisted of torque measurements and lateral displacement tests. External surface area was computed using a 25-µm micro-CT. RESULTS Maximum insertion torque, maximum removal torque, and force levels for displacements were the highest in N1, followed by DT, ST, DC, and SC (α  =  .05). The surface area was largest in DT, followed by N1, ST, DC, and SC. Surface area engaged in cortical bone, however, was the greatest in N1. The surface area of mini-implants had positive correlation with stability. CONCLUSION Among commercial designs, both added tapering and double threading improved stability. N1 was the most stable design within this research design. The new design has the potential to be clinically superior; it has enhanced stability and there is diminished risk of endangering nearby anatomic structures during placement and orthodontic treatment, but the design requires refinements to reduce insertion torque to avoid clinical difficulty and patient discomfort.

Nanodiamond–Gutta Percha Composite Biomaterials for Root Canal Therapy

Root canal therapy (RCT) represents a standard of treatment that addresses infected pulp tissue in teeth and protects against future infection. RCT involves removing dental pulp comprising blood vessels and nerve tissue, decontaminating residually infected tissue through biomechanical instrumentation, and root canal obturation using a filler material to replace the space that was previously composed of dental pulp. Gutta percha (GP) is typically used as the filler material, as it is malleable, inert, and biocompatible. While filling the root canal space with GP is the standard of care for endodontic therapies, it has exhibited limitations including leakage, root canal reinfection, and poor mechanical properties. To address these challenges, clinicians have explored the use of alternative root filling materials other than GP. Among the classes of materials that are being explored as novel endodontic therapy platforms, nanodiamonds (NDs) may offer unique advantages due to their favorable properties, particularly for dental applications. These include versatile faceted surface chemistry, biocompatibility, and their role in improving mechanical properties, among others. This study developed a ND-embedded GP (NDGP) that was functionalized with amoxicillin, a broad-spectrum antibiotic commonly used for endodontic infection. Comprehensive materials characterization confirmed improved mechanical properties of NDGP over unmodified GP. In addition, digital radiography and microcomputed tomography imaging demonstrated that obturation of root canals with NDGP could be achieved using clinically relevant techniques. Furthermore, bacterial growth inhibition assays confirmed drug functionality of NDGP functionalized with amoxicillin. This study demonstrates a promising path toward NDGP implementation in future endodontic therapy for improved treatment outcomes.

Characterization of the osteogenic potential of mesenchymal stem cells from human periodontal ligament based on cell surface markers.

Mesenchymal stem cell (MSC)-mediated therapy has been shown to be clinically effective in regenerating tissue defects. For improved regenerative therapy, it is critical to isolate homogenous populations of MSCs with high capacity to differentiate into appropriate tissues. The utilization of stem cell surface antigens provides a means to identify MSCs from various tissues. However, few surface markers that consistently isolate highly regenerative MSCs have been validated, making it challenging for routine clinical applications and making it all the more imperative to identify reliable surface markers. In this study, we used three surface marker combinations: CD51/CD140α, CD271, and STRO-1/CD146 for the isolation of homogenous populations of dental mesenchymal stem cells (DMSCs) from heterogeneous periodontal ligament cells (PDLCs). Fluorescence-activated cell sorting analysis revealed that 24% of PDLCs were CD51+/CD140α+, 0.8% were CD271+, and 2.4% were STRO-1+/CD146+. Sorted cell populations were further assessed for their multipotent properties by inducing osteogenic and chondrogenic differentiation. All three subsets of isolated DMSCs exhibited differentiation capacity into osteogenic and chondrogenic lineages but with varying degrees. CD271+ DMSCs demonstrated the greatest osteogenic potential with strong induction of osteogenic markers such as DLX5, RUNX2, and BGLAP. Our study provides evidence that surface marker combinations used in this study are sufficient markers for the isolation of DMSCs from PDLCs. These results provide important insight into using specific surface markers for identifying homogenous populations of DMSCs for their improved utilization in regenerative medicine.

Mechanical stability assessment of novel orthodontic mini-implant designs: Part 2

OBJECTIVE To assess the mechanical stability of a newly revised orthodontic mini-implant design (N2) compared with a design introduced in Part 1 of the study (N1) and the most widely-used commercially-available design (CA). To evaluate the mean buccal bone thickness of maxillary and mandibular posterior teeth using cone-beam computed tomography (CBCT). MATERIALS AND METHODS From the CBCT scans of 20 patients, six tomographic cross-sections were generated for each tooth. Buccal bone thickness was measured from the most convex point on the bone to the root surface. CA (1.5 mm in diameter and 6 mm in length), N1, and N2 (shorter and narrower than N1) were inserted in simulated bone with cortical and trabecular bone layers. Mechanical stability was compared in vitro through torque and lateral displacement tests. RESULTS The bone thickness ranged from 2.26 to 3.88 mm. Maximum insertion torque was decreased significantly in N2 compared to N1. However, force levels for all displacement distances and torque ratio were the highest in N2, followed by N1 and CA (α = .05). CONCLUSIONS Both torque and lateral displacement tests highlighted the enhanced stability of N2 compared with CA. Design revisions to N1 effectively mitigated N1s high insertion torque and thus potentially reduced microdamage to the surrounding bone. The N2 design is promising as evidenced by enhanced stability and high mechanical efficiency. Moreover, N2 is not limited to placement in interradicular spaces and has the capacity to be placed in the buccal bone superficial to the root surface with diminished risk of endangering nearby anatomic structures during placement and treatment.

Single CD271 marker isolates mesenchymal stem cells from human dental pulp

Mesenchymal stem cells (MSCs) are a promising tool in regenerative medicine due to their capacity to differentiate into multiple lineages. In addition to MSCs isolated from bone marrow (BMSCs), adult MSCs are isolated from craniofacial tissues including dental pulp tissues (DPs) using various stem cell surface markers. However, there has been a lack of consensus on a set of surface makers that are reproducibly effective at isolating putative multipotent dental mesenchymal stem cells (DMSCs). In this study, we used different combinations of surface markers (CD51/CD140α, CD271, and STRO-1/CD146) to isolate homogeneous populations of DMSCs from heterogeneous dental pulp cells (DPCs) obtained from DP and compared their capacity to undergo multilineage differentiation. Fluorescence-activated cell sorting revealed that 27.3% of DPCs were CD51+/CD140α+, 10.6% were CD271+, and 0.3% were STRO-1+/CD146+. Under odontogenic conditions, all three subsets of isolated DMSCs exhibited differentiation capacity into odontogenic lineages. Among these isolated subsets of DMSCs, CD271+ DMSCs demonstrated the greatest odontogenic potential. While all three combinations of surface markers in this study successfully isolated DMSCs from DPCs, the single CD271 marker presents the most effective stem cell surface marker for identification of DMSCs with high odontogenic potential. Isolated CD271+ DMSCs could potentially be utilized for future clinical applications in dentistry and regenerative medicine.

Transforming Growth Factor-β-Induced KDM4B Promotes Chondrogenic Differentiation of Human Mesenchymal Stem Cells

The high prevalence of cartilage diseases and limited treatment options create a significant biomedical burden. Due to the inability of cartilage to regenerate itself, introducing chondrocyte progenitor cells to the affected site is of significant interest in cartilage regenerative therapies. Tissue engineering approaches using human mesenchymal stem cells (MSCs) are promising due to their chondrogenic potential, but a comprehensive understanding of the mechanisms governing the fate of MSCs is required for precise therapeutic applications in cartilage regeneration. TGF‐β is known to induce chondrogenesis by activating SMAD signaling pathway and upregulating chondrogenic genes such as SOX9; however, the epigenetic regulation of TGF‐β‐mediated chondrogenesis is not understood. In this report, we found that TGF‐β dramatically induced the expression of KDM4B in MSCs. When KDM4B was overexpressed, chondrogenic differentiation was significantly enhanced while KDM4B depletion by shRNA led to a significant reduction in chondrogenic potential. Mechanistically, upon TGF‐β stimulation, KDM4B was recruited to the SOX9 promoter, removed the silencing H3K9me3 marks, and activated the transcription of SOX9. Furthermore, KDM4B depletion reduced the occupancy of SMAD3 in the SOX9 promoter, suggesting that KDM4B is required for SMAD‐dependent coactivation of SOX9. Our results demonstrate the critical role of KDM4B in the epigenetic regulation of TGF‐β‐mediated chondrogenic differentiation of MSCs. Since histone demethylases are chemically modifiable, KDM4B may be a novel therapeutic target in cartilage regenerative therapy. Stem Cells 2016;34:711–719