Periodontal Phenotype Modification Using Subepithelial Connective Tissue Graft and Bone Graft in the Mandibular Anterior Teeth with Mucogingival Problems Following Orthodontic Treatment.

Among the complications of orthodontic treatment, mucogingival problems with gingival recession in the mandibular anterior teeth are challenging for clinicians. Mucogingival problems can lead to esthetic deficits, thermal hypersensitivity, tooth brushing pain, and complicated plaque control. Herein, we present a case of a 16-year-old female with gingival recession in the left mandibular central incisor after orthodontic treatment. The preoperative clinical findings showed a thin soft tissue biotype with root prominence in the mandibular anterior area. The interdental area was relatively depressed. After reflection of the full-thickness flap, root coverage using a bone graft substitute and subepithelial connective tissue graft obtained from the palatal mucosa was performed. The 6-month and 5-year postoperative clinical findings showed improved soft tissue phenotype. The cross-sectional CBCT scans 5 years after surgery showed a well-maintained labial bone plate in the mandibular incisors. Within the limitations of this case report, for patients with gingival recession in the mandibular incisors after orthodontic treatment, a successful biotype modification can be achieved with a combined procedure using subepithelial connective tissue graft with bone graft substitutes.

Use of a Lateral Sinus Bony Window as an Intraoral Donor Site for Guided Bone Regeneration in Wide Post-Extraction Defects.

Maxillary sinus augmentation (MSA) and guided bone regeneration (GBR) have shown successful clinical, radiological, and histological outcomes for implant-related bone reconstruction and have been used to augment bony defects of various shapes and sizes. This study demonstrated that the lateral sinus bony window obtained during MSA can be used as an autogenous block bone graft for the augmentation of wide post-extraction defects. During the uncovering procedure performed 6 months after surgery, the grafted lateral bony window was well integrated with the adjacent native bone, and complete bone filling was observed in all bony defects around the implants. All of the implants survived. Within the limitations of this study, autogenous block bone obtained from lateral window sites can be used as novel donors for the resolution of wide bony defects around implants.

Novel Osteogenic Behaviors around Hydrophilic and Radical-Free 4-META/MMA-TBB: Implications of an Osseointegrating Bone Cement.

Poly(methyl methacrylate) (PMMA)-based bone cement, which is widely used to affix orthopedic metallic implants, is considered bio-tolerant but lacks osteoconductivity and is cytotoxic. Implant loosening and toxic complications are significant and recognized problems. Here we devised two strategies to improve PMMA-based bone cement: (1) adding 4-methacryloyloxylethyl trimellitate anhydride (4-META) to MMA monomer to render it hydrophilic; and (2) using tri-n-butyl borane (TBB) as a polymerization initiator instead of benzoyl peroxide (BPO) to reduce free radical production. Rat bone marrow-derived osteoblasts were cultured on PMMA-BPO, common bone cement ingredients, and 4-META/MMA-TBB, newly formulated ingredients. After 24 h of incubation, more cells survived on 4-META/MMA-TBB than on PMMA-BPO. The mineralized area was 20-times greater on 4-META/MMA-TBB than PMMA-BPO at the later culture stage and was accompanied by upregulated osteogenic gene expression. The strength of bone-to-cement integration in rat femurs was 4- and 7-times greater for 4-META/MMA-TBB than PMMA-BPO during early- and late-stage healing, respectively. MicroCT and histomorphometric analyses revealed contact osteogenesis exclusively around 4-META/MMA-TBB, with minimal soft tissue interposition. Hydrophilicity of 4-META/MMA-TBB was sustained for 24 h, particularly under wet conditions, whereas PMMA-BPO was hydrophobic immediately after mixing and was unaffected by time or condition. Electron spin resonance (ESR) spectroscopy revealed that the free radical production for 4-META/MMA-TBB was 1/10 to 1/20 that of PMMA-BPO within 24 h, and the substantial difference persisted for at least 10 days. The compromised ability of PMMA-BPO in recruiting cells was substantially alleviated by adding free radical-scavenging amino-acid N-acetyl cysteine (NAC) into the material, whereas adding NAC did not affect the ability of 4-META/MMA-TBB. These results suggest that 4-META/MMA-TBB shows significantly reduced cytotoxicity compared to PMMA-BPO and induces osteoconductivity due to uniquely created hydrophilic and radical-free interface. Further pre-clinical and clinical validations are warranted.

Tuning of Titanium Microfiber Scaffold with UV-Photofunctionalization for Enhanced Osteoblast Affinity and Function.

Titanium (Ti) is an osteoconductive material that is routinely used as a bulk implant to fix and restore bones and teeth. This study explored the effective use of Ti as a bone engineering scaffold. Challenges to overcome were: (1) difficult liquid/cell infiltration into Ti microfiber scaffolds due to the hydrophobic nature of Ti; and (2) difficult cell attachment on thin and curved Ti microfibers. A recent discovery of UV-photofunctionalization of Ti prompted us to examine its effect on Ti microfiber scaffolds. Scaffolds in disk form were made by weaving grade 4 pure Ti microfibers (125 µm diameter) and half of them were acid-etched to roughen the surface. Some of the scaffolds with original or acid-etched surfaces were further treated by UV light before cell culture. Ti microfiber scaffolds, regardless of the surface type, were hydrophobic and did not allow glycerol/water liquid to infiltrate, whereas, after UV treatment, the scaffolds became hydrophilic and immediately absorbed the liquid. Osteogenic cells from two different origins, derived from the femoral and mandibular bone marrow of rats, were cultured on the scaffolds. The number of cells attached to scaffolds during the early stage of culture within 24 h was 3-10 times greater when the scaffolds were treated with UV. The development of cytoplasmic projections and cytoskeletal, as well as the expression of focal adhesion protein, were exclusively observed on UV-treated scaffolds. Osteoblastic functional phenotypes, such as alkaline phosphatase activity and calcium mineralization, were 2-15 times greater on UV-treated scaffolds, with more pronounced enhancement on acid-etched scaffolds compared to that on the original scaffolds. These effects of UV treatment were associated with a significant reduction in atomic carbon on the Ti microfiber surfaces. In conclusion, UV treatment of Ti microfiber scaffolds tunes their physicochemical properties and effectively enhances the attachment and function of osteoblasts, proposing a new strategy for bone engineering.

Effect of UV Photofunctionalization on Osseointegration in Aged Rats.

OBJECTIVES: This study evaluated the effect of photofunctionalization on osseointegration under the biologically adverse conditions of aging. MATERIALS: First of all, bone marrow-derived osteoblastic cells from young (8 weeks old) and aged (15 months old) rats were biologically characterized. Then, the osteoblasts from aged rats were seeded on titanium discs with and without photofunctionalization, and assessed for initial cell attachment and osteoblastic functions. Titanium mini-implants, with and without photofunctionalization, were placed in the femur of aged rats, and the strength of osseointegration was measured at week 2 of healing. Periimplant tissue was examined morphologically and chemically using scanning electron microscopy and energy dispersive x-ray spectroscopy, respectively. RESULTS: Cells from the aged rats showed substantially reduced biological capabilities compared with those derived from young rats. The cells from aged rats showed significantly increased cell attachment and the expression of osteoblastic function on photofunctionalized titanium than on untreated titanium. In addition, the strength of osseointegration was increased by 40% in aged rats carrying the photofunctionalized implants. Robust bone formation was observed around the photofunctionalized implants with strong elemental peaks of calcium and phosphorus, whereas the tissue around untreated implants showed weaker calcium and phosphate signals than titanium ones. CONCLUSION: These in vivo and in vitro results corroboratively demonstrate that photofunctionalization is effective for enhancing osseointegration in aged rats.

Bone Generation Profiling Around Photofunctionalized Titanium Mesh.

PURPOSE: The aim of this study was to evaluate whether photofunctionalization of titanium mesh enhances its osteoconductive capability. MATERIALS AND METHODS: The titanium mesh (0.2 mm thickness) used in this study was made of commercially pure grade-2 titanium and had hexagonal apertures (2 mm width). Photofunctionalization was performed by treating titanium mesh with UV light for 12 minutes using a photo device immediately before use. Untreated or photofunctionalized titanium mesh was placed into rat femurs, and bone generation around titanium mesh was profiled using three-dimensional (3D) microcomputed tomography (micro-CT). A set of in vitro experiments was conducted using bone marrow-derived osteoblasts. RESULTS: Photofunctionalized titanium mesh surfaces were characterized by the regenerated hydrophilicity and significantly reduced surface carbon. Bone generation profiling at week 3 of healing showed that the hexagonal apertures in photofunctionalized mesh were 95% filled, but they were only 57% filled in untreated mesh, particularly with the center zone remaining as a gap. Bone profiling in slices parallel to the titanium surface showed that photofunctionalized titanium mesh achieved 90% bone occupancy 0 to 400 µm from the surface, compared with only 35% for untreated mesh. Bone occupancy remained as high as 55% 800 to 1,200 µm from photofunctionalized titanium mesh surfaces, compared with less than 20% for untreated mesh. In vitro, photofunctionalized titanium mesh expedited and enhanced attachment and spread of osteoblasts, and increased ALP activity and the rate of mineralization. CONCLUSION: This study may provide novel and advanced metrics describing the osteoconductive property of photofunctionalized titanium mesh. Specifically, photofunctionalization not only increased the breadth, but also the 3D range, of osteoconductivity of titanium mesh, enabling space-filling and far-reaching osteoconductivity. Further translational and clinical studies are warranted to establish photofunctionalized titanium mesh as a novel clinical tool for better bone regeneration and augmentation.

Engineering bone-implant integration with photofunctionalized titanium microfibers.

There are significant challenges in regenerating large volumes of bone tissue, and titanium implant therapy is extremely difficult or contraindicated when there is no supporting bone. Surface conditioning of titanium implants with UV light immediately prior to use, or photofunctionalization, improves the speed and degree of bone-implant integration. Here, we hypothesized that photofunctionalized titanium microfibers are capable of promoting bone ingrowth into the microfiber scaffold to improve bone-implant integration in bone defects. Titanium implants (1 mm in diameter, 2 mm in length) enfolded with 0.7 mm-thick titanium microfibers were placed into 2.4-mm diameter osteotomy in rat femurs. Titanium microfibers and implants were photofunctionalized by treatment with UV light for 12 min using a photo device immediately prior to surgery. Photofunctionalized microfibers and implants were hydrophilic, while as-made microfiber-enfolded implants were hydrophobic. Implant anchorage strength was 2.5 times and 2.2 times greater for photofunctionalized microfiber-enfolded implants than as-made ones at weeks 2 and 4 of healing, respectively. Robust bone formation was only seen at the implant surface of photofunctionalized microfiber-enfolded implants. Bone formation as measured by the Ca/Ti ratio was 5 to over 20 times greater for photofunctionalized than as-made microfiber scaffolds. The Ca/P ratio was 1.55-1.65 in the tissue produced in photofunctionalized microfibers and 1.1-1.3 in tissue in as-made microfibers. In vitro, the number of attached osteoblasts and their alkaline phosphatase activity, both near zero on as-received microfibers, were significantly increased on photofunctionalized microfibers. In conclusion, bone ingrowth occurred in photofunctionalized titanium microfiber scaffolds, enabling successful bone-implant integration when the microfiber-enfolded implants were placed in a site without primary bone support. The combined use of titanium microfibers and photofunctionalization may provide a novel and effective strategy to regenerate and integrate bone in a wider range of applications.

Effect of Photofunctionalization on Ti6Al4V Screw Stability Placed in Segmental Bone Defects in Rat Femurs.

PURPOSE: Ultraviolet-mediated photofunctionalization is a new technology to improve bone and titanium integration. We hypothesized that photofunctionalization would enhance the stability of titanium screws used for segmental bone defects. MATERIALS AND METHODS: Disks were prepared of a titanium alloy (Ti6Al4V) for an in vitro study to evaluate the attachment, proliferation, and differentiation of osteoblasts. Commercially available Ti6Al4V screws were used in vivo. Segmental bone defects were created in rat femurs as an immediate loading reconstruction model. The defects were reconstructed with commercially available titanium plates and Ti6Al4V screws, with or without photofunctionalization. The screw survival rates and mechanical stability were evaluated at 2 and 4 weeks, and the bone formation around the screws was analyzed. RESULTS: Osteoblasts showed greater attachment, proliferation, and differentiation on the photofunctionalized Ti6Al4V disks. Photofunctionalized screws had significantly greater survival rates and mechanical stability at 2 and 4 weeks. The bone formation around the photofunctionalized screws was significantly greater than that around the untreated screws at 4 weeks. CONCLUSIONS: The results of the present study have demonstrated the efficacy of photofunctionalization on enhancing the survival and stability of Ti6Al4V screws under a loaded condition in the reconstruction of segmental defects. This was associated with increased bioactivity and bone formation around the photofunctionalized Ti6Al4V material.

Effect of UV Photofunctionalization on Biologic and Anchoring Capability of Orthodontic Miniscrews.

PURPOSE: Treatment of titanium with UV light immediately before use, or photofunctionalization, is gaining traction as a simple method to improve the biologic capability and clinical performance of dental implants. The objective of this study was to examine the effect of photofunctionalization on the biologic capability and mechanical anchorage of orthodontic miniscrews. MATERIALS AND METHODS: Untreated and photofunctionalized Ti-6Al-4V orthodontic miniscrews were placed into rat femurs. Photofunctionalization was performed by treating miniscrews with UV light for 12 minutes using a photo device immediately before placement. After 3 weeks of healing, miniscrews were pushed laterally to measure the resistance against the tipping force. The miniscrews were also evaluated for morphology and chemistry of tissue formed around them using scanning electron microscopy and energy dispersive spectroscopy. Rat bone marrow-derived osteoblasts were cultured on Ti-6Al-4V disks with and without photofunctionalization. The number of osteoblasts attached to the disks and the behaviors, alkaline phosphatase activity, and mineralization capability of osteoblasts were evaluated. RESULTS: Photofunctionalization converted both disk and screw surfaces from hydrophobic to superhydrophilic. In vivo biomechanical testing showed that the displacement of untreated screws was 1.5 to 1.7 times greater than that of photofunctionalized screws when subjected to lateral tipping force. Robust bone formation was observed around photofunctionalized miniscrews with strong elemental peaks of calcium and phosphorus, whereas the tissue around untreated miniscrews appeared thin and showed no clear peak of calcium. The attachment, initial spreading, adhesion, and expression of functional phenotypes of osteoblasts were significantly increased on photofunctionalized Ti-6Al-4V disks. CONCLUSION: These in vivo and in vitro results comprehensively and consistently demonstrate that photofunctionalization increases the bioactivity of Ti-6Al-4V and improves the anchoring capability of orthodontic miniscrews.

Ultraviolet photofunctionalization increases removal torque values and horizontal stability of orthodontic miniscrews.

INTRODUCTION: The objective of this study was to examine the effects of ultraviolet-mediated photofunctionalization of miniscrews and the in-vivo potential of bone-miniscrew integration. METHODS: Self-drilling orthodontic miniscrews made from a titanium alloy were placed in rat femurs. Photofunctionalization was performed by treating the miniscrews with ultraviolet light for 12 minutes with a photo device immediately before implantation. Maximum insertion torque (week 0), removal torque (weeks 0 and 3), and resistance to lateral tipping force (week 3) were examined. RESULTS: The removal torque at 3 weeks of healing was higher for the photofunctionalized screws than for the untreated screws. The regenerated bone tissue was more intact and contiguous around the photofunctionalized miniscrews than around the untreated ones. The miniscrew-bone complex seemed to produce interface failure, not cohesive fracture, in both groups. The displacement of untreated screws under a lateral tipping force was greater than that of photofunctionalized miniscrews. CONCLUSIONS: These results suggest that photofunctionalization increases the bioactivity of titanium-alloy miniscrews and improves the anchoring capability of orthodontic miniscrews, even without modification of the surface topography.

Evidence for novel structures (primo vessels and primo nodes) floating in the venous sinuses of rat brains.

We for the first time report evidence for existence of novel structures, primo vessels (PVs) and primo nodes (PNs) floating inside the venous sinuses of rat brains. For this purpose, we applied a chromium-hematoxylin (Cr-Hx) solution to stain the PVs and the PNs floating inside the venous sinuses (superior sagittal sinus, strait sinus, and transverse sinus) of seven rats' brains preferentially compared to the blood clots that easily form during surgery. Cr-Hx-stained PVs and PNs were examined by light and transmission electron microscopies: (1) we were consistently able to visualize the PVs and the PNs in the venous sinuses of all seven rats' brains. (2) The PVs and PNs consisted of rod-shaped and some round-shaped cells, respectively, as demonstrated by using 4',6-diamidino-2-phenylindole (DAPI). (3) Cross sections of the PVs showed that the sinuses contained loose fibrous materials and clusters of nano-sized granules enveloped by the cortex. The above data imply that thrombus of the venous sinuses may be related with these novel floating structures. However, the functions of the PVs and PNs floating in the venous sinuses remain to be investigated in terms of normal or thrombus-provoked diseases.

4 Hz magnetic field decreases oxidative stress in mouse brain: a chemiluminescence study.

We investigated the effects of delta and theta brain wave frequency magnetic fields (3, 4, and 5) on mouse brain by detecting photonic oxidative stress makers; spontaneous photon emission (SPE) and lucigenin and tert-butyl hydroperoxide (TBHP) induced chemiluminescences (CL). For this purpose, Balb/C mice were exposed to 3, 4, and 5 Hz magnetic fields (MF) at 0.7 mT for 3 h, respectively. After that we monitored SPE and lucigenin and TBHP-induced CL of the homogenates of mice brains. There was a significant decrease in SPE in the 4 Hz MF-exposed group. Lucigenin-induced CL was also significantly decreased only in the 4 Hz MF-exposed group. TBHP-induced CL was also distinctively decreased by all frequencies, 3, 4, and 5 Hz MF exposures. These results showed that oxidative stress in a mouse brain was decreased by 4 Hz MF. We suggest that the application of 4 Hz MF will contribute to magnetic field therapy.