Wetting of Cell Aggregates on Microdisk Topography Structures Achieved by Maskless Optical Projection Lithography.

Cell aggregates as a 3D culture model can effectively mimic the physiological processes such as embryonic development, immune response, and tissue renewal in vivo. Researches show that the topography of biomaterials plays an important role in regulating cell proliferation, adhesion, and differentiation. It is of great significance to understand how cell aggregates respond to surface topography. Herein, microdisk array structures with the optimized size are used to investigate the wetting of cell aggregates. Cell aggregates exhibit complete wetting with distinct wetting velocities on the microdisk array structures of different diameters. The wetting velocity of cell aggregates reaches a maximum of 293 µm h-1 on microdisk structures with a diameter of 2 µm and is a minimum of 247 µm h-1 on microdisk structures of 20 µm diameter, which suggests that the cell-substrates adhesion energy on the latter is smaller. Actin stress fibers, focal adhesions (FAs), and cell morphology are analyzed to reveal the mechanisms of variation of wetting velocity. Furthermore, it is demonstrated that cell aggregates adopt climb and detour wetting modes on small and large-sized microdisk structures, respectively. This work reveals the response of cell aggregates to micro-scale topography, providing guidance for better understanding of tissue infiltration.

Exosomes Derived from Adipose Tissue-Derived Mesenchymal Stromal Cells Prevent Medication-Related Osteonecrosis of the Jaw through IL-1RA.

Medication-related osteonecrosis of the jaw (MRONJ) is a severe disease with unclear pathogenesis. Adipose tissue-derived mesenchymal stromal cells (MSC(AT)s) serve as a special source for cell therapy. Herein, we explored whether exosomes (Exo) derived from MSC(AT)s promote primary gingival wound healing and prevent MRONJ. An MRONJ mice model was constructed using zoledronate (Zol) administration and tooth extraction. Exosomes were collected from the conditioned medium (CM) of MSC(AT)s (MSC(AT)s-Exo) and locally administered into the tooth sockets. Interleukin-1 receptor antagonist (IL-1RA)-siRNA was used to knock down the expression of IL-1RA in MSC(AT)s-Exo. Clinical observations, micro-computed tomography (microCT), and histological analysis were used to evaluate the therapeutic effects in vivo. In addition, the effect of exosomes on the biological behavior of human gingival fibroblasts (HGFs) was evaluated in vitro. MSC(AT)s-Exo accelerated primary gingival wound healing and bone regeneration in tooth sockets and prevented MRONJ. Moreover, MSC(AT)s-Exo increased IL-1RA expression and decreased interleukin-1 beta (IL-1ß) and tumor necrosis factor-α (TNF-α) expression in the gingival tissue. The sequent rescue assay showed that the effects of preventing MRONJ in vivo and improving the migration and collagen synthesis abilities of zoledronate-affected HGFs in vitro were partially impaired in the IL-1RA-deficient exosome group. Our results indicated that MSC(AT)s-Exo might prevent the onset of MRONJ via an IL-1RA-mediated anti-inflammatory effect in the gingiva wound and improve the migration and collagen synthesis abilities of HGFs.

Low-level laser therapy prevents medication-related osteonecrosis of the jaw-like lesions via IL-1RA-mediated primary gingival wound healing.

BACKGROUND: Medication-related osteonecrosis of the jaw (MRONJ) is a serious debilitating disease caused by anti-resorption and anti-angiogenesis drugs, significantly affecting patients' quality of life. Recent studies suggested that primary gingival wound healing may effectively prevent the development of MRONJ. This study aimed to evaluate the effects of low-level light therapy (LLLT) on promoting gingival wound healing in extraction sockets of MRONJ-like mice and preventing the occurrence of MRONJ. Furthermore, we explored underlying mechanisms. METHODS: Mice were randomly divided into the Ctrl, Zol, and Zol + LLLT groups. Administration of zoledronate and tooth extraction of bilateral maxillary second molars were used to build the MRONJ model, and LLLT was locally administered into the tooth sockets to examine the effect of LLLT. Next, to explore the function of IL-1RA, we performed LLLT with interleukin-1 receptor antagonist (IL-1RA) neutralizing antibody (named Zol + LLLT + IL-1RA NAb group) or negative control antibodies for tooth extraction in subsequent rescue animal experiments. Stereoscope observations, micro-computed tomography, and histological examination were conducted to evaluate gingival wound healing and bone regeneration in tooth sockets. The effects of LLLT on the migration capacities of zoledronate-treated epithelial cells were assessed in vitro. RESULTS: LLLT promoted primary gingival wound healing without exposed necrotic bone. Micro-computed tomography results showed higher bone volume and mineral density of the tooth sockets after LLLT. Histology analysis showed complete gingival coverage, obvious bone regeneration, and reduced soft tissue inflammation, with down-regulated pro-inflammation cytokines, like interleukin-1 beta (IL-1ß) and tumor necrosis factor-α (TNF-α), and up-regulated IL-1RA expression in the gingival tissue in the LLLT group. The rescue assay further showed that the effects of LLLT promoting gingival wound healing and preventing MRONJ might be partially abolished by IL-1RA neutralizing antibodies. In vitro studies demonstrated that LLLT accelerated zoledronate-treated epithelial cell migration. CONCLUSIONS: LLLT might promote primary gingival wound healing and contribute to subsequent bone regeneration of the tooth extractions in MRONJ-like lesions via IL-1RA-mediated pro-inflammation signaling suppression.

Exosomes from Adipose-Derived Stem Cells Can Prevent Medication-Related Osteonecrosis of the Jaw.

The treatment measures of medication-related osteonecrosis of the jaw (MRONJ) is a worldwide challenge in oral and maxillofacial surgery because of its unclear pathogenesis. Previous studies suggested that mesenchymal stem cells played important roles in promoting MRONJ lesion healing, but the detailed mechanisms were unknown. Increasing numbers of studies have demonstrated that exosomes derived from mesenchymal stem cells, especially adipose-derived stem cells, have key roles in stem cell-based therapies by accelerating bone remodeling, facilitating angiogenesis, and promoting wound healing. We hypothesized that exosomes derived from adipose-derived stem cells can prevent MRONJ by accelerating gingival healing and enhancing bone remodeling processes. Our results may provide a promising therapeutic option for MRONJ clinical therapy.

A systematic review of the accuracy of digital surgical guides for dental implantation.

PURPOSE: This review aimed to reveal the influence of implant guides on surgical accuracy with regard to supporting types, manufacturing methods and design (including fixation screws and sleeves). METHODS: A literature search related to accuracy of surgical guides for dental implantation was performed in Web of Science and PubMed. Studies with in vivo or in vitro deviation data published in recent 5 years (2018-2022) were included and assessed by Newcastle-Ottawa Scale with regard to risk of bias and reliability degree of clinical studies. Accuracy-related deviation data were summarized as forest plots and normal distributions. RESULTS: Forty-one articles were included with high degree of credibility. Data showed that implant surgery accuracy can be achieved with mean distance deviation < 2 mm (most < 1 mm) and angular deviation < 8° (most < 5°). CONCLUSIONS: Bilateral tooth-supported guides exhibited highest in vitro accuracy and similar in vivo accuracy to unilateral tooth-supported guides; mucosa-supported guides exhibit lowest in vivo accuracy, while its in vitro data showed low credibility due to mechanical complexity of living mucosa tissue. Milling exhibited higher in vivo accuracy of guides than 3d-printing, though further data support was needed. Design of fixation screws and sleeves of implant guides affected the surgical accuracy and might remain a research focus in near future. However, lack of universal evaluation standards for implantation accuracy remained a major problem in this field. The influence of implant guides on surgical accuracy revealed in this review might shed light on future development of dental implantology.

Increased apoptosis of gingival epithelium is associated with impaired autophagic flux in medication-related osteonecrosis of the jaw.

Macroautophagy/autophagy has both negative and positive aspects in the development of many diseases. Yet, its exact role and specific mechanism in the onset of medication-related osteonecrosis of the jaw (MRONJ) is still not fully understood. Retarded gingiva healing is the primary clinical manifestation in patients with MRONJ. In this study, we aimed to explore the relationship between autophagy and apoptosis in MRONJ gingival epithelium and search for a method to prevent this disease. First, we examined clinical samples from patients diagnosed with MRONJ and healthy controls, finding that autophagy-related markers MAP1LC3/LC3 and SQSTM1/p62 synchronously increased, thus suggesting that autophagic flux was suppressed in MRONJ. Moreover, mRNA sequencing analysis and TUNEL assay showed that the process of apoptosis was upregulated in patients and animals with MRONJ, indicating autophagy and apoptosis participate in the development of MRONJ. Furthermore, the level of autophagy and apoptosis in zoledronic acid (ZA)-treated human keratinocytes cell lines (HaCaT cells) was concentration dependent in vitro. In addition, we also found that RAB7 (RAB7, member RAS oncogene family) activator ML098 could rescue MRONJ gingival lesions in mice by activating the autophagic flux and downregulating apoptosis. To sum up, this study demonstrated that autophagic flux is impaired in the gingival epithelium during MRONJ, and the rescued autophagic flux could prevent the occurrence of MRONJ.Abbreviations: ACTB: actin beta; Baf-A1: bafilomycin A1; CASP3: caspase 3; CASP8: caspase 8; CT: computed tomography; DMSO: dimethyl sulfoxide; GFP: green fluorescent protein; HaCaT cells: human keratinocytes cell lines; H&E: hematoxylin and eosin; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MRONJ: medication-related osteonecrosis of the jaw; PARP: poly(ADP-ribose) polymerase; RAB7: RAB7, member RAS oncogene family; RFP: red fluorescent protein; SQSTM1/p62: sequestosome 1; TEM: transmission electron microscopy; ZA: zoledronic acid.

[Consistency of the landmark identification on two types of cephalograms from cone-beam CT].

PURPOSE: To study the landmark consistency on two types of cephalograms reconstructed from the data of cone-beam CT(CBCT). METHODS: CBCT data of 56 patients without teeth missing were uploaded into Dolphin software and transferred to the cephalograms. In the control group, the right half projection was taken while in the experimental group, the images achieved after 6 individual projections were managed to achieve the synthesized cephalograms. One experienced orthodontists traced the 23 frequently-used landmarks and the consistency was compared by means of Hotelling T2 test with SPSS 13.0 software package. RESULTS: The tracing of anterior nasal ridge, posterior nasal ridge, upper incisor root tip, lower incisor edge, lower incisor root tip and condyle showed significant difference(P<0.05). CONCLUSIONS: The synthesized cephalograms in the experimental group show the anatomical structure more precisely which is more beneficial to landmark identification.

Adipose-Derived Stem Cells Promote Bone Coupling in Bisphosphonate-Related Osteonecrosis of the Jaw by TGF-ß1.

This study aimed to investigate molecularly targeted therapy to revive bone remodeling and prevent BRONJ by local adipose-derived stem cells (ADSCs) transplantation. Clinical samples of BRONJ and healthy jawbones were used to examine the bone coupling-related cells and TGF-ß1 expression. Bone coupling-related cells and TGF-ß1 expression were also assessed in BRONJ-like animal model to confirm the results in clinical samples. ADSCs were locally administered in vivo and the therapeutic effects were evaluated by gross observation, radiological imaging, and histological examination. Furthermore, ADSCs-conditioned medium (ADSCs-CM) and neutralizing antibody were applied to assess the effects of ADSCs-derived TGF-ß1 on restoring bone coupling in vivo. Osteoclast formation and resorption assays were performed to evaluate the effects of ADSCs-derived TGF-ß1 on ZA-treated pre-osteoclasts. Cell migration was performed to assess the effects of ADSCs-derived TGF-ß1 on patients' bone marrow stem cells (BMSCs). The number of osteoclasts, Runx2-positive bone-lining cells (BLCs) and TGF-ß1 expression were decreased in BRONJ and animal model jaw bone samples. These reductions were significantly rescued and necrotic jawbone healing was effectively promoted by local ADSCs administration in BRONJ-like animal models. Mechanistically, ADSCs-CM mainly contributed to promoting bone coupling, while TGF-ß1 neutralizing antibody in the conditioned medium inhibited these effects. Besides, osteoclastogenesis and patients' BMSCs migration were also rescued by ADSCs-derived TGF-ß1. Furthermore, bone resorption-released bone matrix TGF-ß1, together with ADSCs-derived TGF-ß1, synergistically contributed to rescuing BMSCs migration. Collectively, ADSCs promoted bone healing of BRONJ by TGF-ß1-activated osteoclastogenesis and BMSCs migration capacities.

Cucurbit[7]uril-Carbazole Two-Photon Photoinitiators for the Fabrication of Biocompatible Three-Dimensional Hydrogel Scaffolds by Laser Direct Writing in Aqueous Solutions.

We have introduced a novel water-soluble two-photon photoinitiator based on the host-guest interaction between 3,6-bis[2-(1-methyl-pyridinium)vinyl]-9-pentyl-carbazole diiodide (BMVPC) and cucurbit[7]uril (CB7) because most of the commercial photoinitiators have poor two-photon initiating efficiency in aqueous solutions. The binding ratio of BMVPC and CB7 was determined as 1:1 by isothermal titration calorimetry and quantum chemical calculation. The formation of the host-guest complex increases the two-photon absorption cross-section about five times, and improves the water solubility required as the photoinitiator for hydrogel fabrication. The BMVPC-CB7 inclusion complex was used as the one-component photoinitiator, and the polyethylene glycol diacrylate with promising biocompatibility was used as the hydrogel monomer to form the aqueous-phase photoresist system applied to two-photon polymerization microfabrication. A relatively low laser threshold of 4.5 mW, a high fabricating resolution of 180 nm, and the true three-dimensional (3D) fabricating capability in the aqueous solution have been obtained by using the as-prepared photoresist system. Finally, 3D engineering hydrogel scaffold microstructures with low toxicity and good biocompatibility have been fabricated and cocultured with living HeLa cells, which demonstrates the potential for further application in tissue engineering.

[Stress distribution in abutment teeth and related tissues under different design of connector: three-dimensional finite element analysis].

PURPOSE: To compare the stress distribution in abutment teeth and related tissues under the same material and different loading between improved major connector design and traditional major connector design. METHODS: One 55-year-old male patient with unilateral maxillary first molar and second molar missing was chosen. The stress distribution in abutment teeth and related tissues were evaluated with spiral CT scanning, Mimics, Geomagic Studio software, a study model was built and finite element analysis was performed using ANSYS software. RESULTS: With the improved major connector design, the stress of abutment decreased significantly, the stress of periodontal decreased, the stress of edentulous mucosa increased significantly and became more balanced, the trend of stimulated absorption of alveolar bone decreased. CONCLUSIONS: For patients with distal free defect of dentition, the design of improved major connector has the effect of stress interruption, can protect the abutment better, detract the stress of the denture and has an good protective effect on the edentulous mucosa and alveolar bone.

The influence of obturators on the respiration of patients with maxillary defects: a clinical study.

The study evaluated the effects of obturators on respiratory function by analyzing the changes in nasal anatomic structures and physiologic function in maxillectomy patients with and without obturators. Twenty-six patients who underwent maxillectomy were chosen and rehabilitated with obturators by a single maxillofacial prosthodontist. The geometric shape of the nasal cavity, the nasal airway resistance, and the ratio of residual volume to total lung capacity (RV/TLC) were evaluated using acoustic rhinometry, rhinomanometry, and a pulmonary function test apparatus, respectively. All patients were tested twice, with and without their obturators. The results were statistically analyzed with a paired t-test. The nasal cavities (0-7 cm to the anterior nostril) of the patients with obturators had a significantly smaller volume ([-8.92, -0.60], P = 0.027), smaller effective nasal cross-sectional area MCA2 ([-3.80, -1,81], P < 0.0001), increased airflow in the nasal cavity ([17.76, 147.39], P = 0.015), reduced nasal airway resistance ([-0.11, -0.02], P = 0.009), and reduced RV/TLC ([-5.32, -1.30], P = 0.004) compared with the patients without obturators. According to the results of this study, obturators can improve respiratory function by effectively decreasing the volume of enlarged nasal cavities as well as the nasal air resistance and volume of anatomical dead space after maxillectomy.

Rehabilitation of maxillectomy defects with obturator prostheses fabricated using computer-aided design and rapid prototyping: a pilot study.

PURPOSE: To establish an alternative method to design and fabricate an obturator prosthesis within the maxillectomy defect using a computer-aided design (CAD) and rapid prototyping (RP) technique and to evaluate the functional results of this technique. MATERIALS AND METHODS: Eleven patients with acquired maxillary defects resulting from head and neck cancers were treated using a protocol based on three-dimensional (3D) reconstruction, CAD, and RP technologies to fabricate obturator prostheses. To evaluate the quality of the obturator prostheses and the patients' satisfaction, the Obturator Functioning Scale (OFS) of the Memorial Sloan-Kettering Cancer Center was applied. RESULTS: Each patient received an individualized obturator that exactly matched the static shape and fit of the defect. Clinical modifications were required to improve border contours. The patients showed good results in all fields of functional outcomes and social acceptance. The OFS scores were comparable with those reported in other studies using traditional maxillectomy impression methods. CONCLUSIONS: This study combined CAD with RP technology to explore an alternative and feasible method for manufacturing individualized obturators for patients after maxillary resection. It has shown significant clinical value, especially for use in developing countries.

A systematic review of visual and instrumental measurements for tooth shade matching.

OBJECTIVE: Common methods for matching tooth shade can be classified into two categories: visual and instrumental measurements. This systematic review evaluated these two methods in terms of precision and accuracy using the agreement percentage rate. METHOD AND MATERIALS: The following databases were searched for studies comparing different shade-matching methods: MEDLINE (PubMed), Embase, and Google Scholar. Reference lists of relevant articles were also searched. Screening, data abstraction, and quality assessment were conducted independently and in duplicate. RESULTS: Twenty-six studies were included. Seventeen studies provided data on the precision of the evaluated shade-matching methods, eight provided data on accuracy, and one provided data on both precision and accuracy. CONCLUSION: Instrumental measurements using a spectrophotometer may provide the most precise and accurate shade-matching outcomes. However, the limitations of the available articles suggest that high-quality studies are still needed verify this claim.