UV Light-Generated Superhydrophilicity of a Titanium Surface Enhances the Transfer, Diffusion and Adsorption of Osteogenic Factors from a Collagen Sponge.

It is a significant challenge for a titanium implant, which is a bio-inert material, to recruit osteogenic factors, such as osteoblasts, proteins and blood effectively when these are contained in a biomaterial. The objective of this study was to examine the effect of ultraviolet (UV)-treatment of titanium on surface wettability and the recruitment of osteogenic factors when they are contained in an atelocollagen sponge. UV treatment of a dental implant made of commercially pure titanium was performed with UV-light for 12 min immediately prior to the experiments. Superhydrophilicity on dental implant surfaces was generated with UV-treatment. The collagen sponge containing blood, osteoblasts, or albumin was directly placed on the dental implant. Untreated implants absorbed only a little blood from the collagen sponge, while the UV-treated implants absorbed blood rapidly and allowed it to spread widely, almost over the entire implant surface. Blood coverage was 3.5 times greater for the UV-treated implants (p < 0.001). Only 6% of the osteoblasts transferred from the collagen sponge to the untreated implants, whereas 16% of the osteoblasts transferred to the UV-treated implants (p < 0.001). In addition, a weight ratio between transferred albumin on the implant and measured albumin adsorbed on the implant was 17.3% in untreated implants and 38.5% in UV-treated implants (p < 0.05). These results indicated that UV treatment converts a titanium surface into a superhydrophilic and bio-active material, which could recruite osteogenic factors even when they were contained in a collagen sponge. The transfer and subsequent diffusion and adsorption efficacy of UV-treated titanium surfaces could be useful for bone formation when titanium surfaces and osteogenic factors are intervened with a biomaterial.

Does local injection of reveromycin A inhibit tooth movement without causing systemic side effects?

OBJECTIVE: To determine the feasibility of local inhibition of osteoclast activity and control of tooth movement with local intraoral reveromycin A (RMA) injection in model mice for experimental tooth movement. MATERIALS AND METHODS: Eight-week-old wild-type mice (n = 6 per group) were divided into four groups consisting of two non-RMA groups that received normal saline for 14 (14-day non-RMA group) or 21 consecutive days (21-day non-RMA group) and 2 RMA groups that received RMA (1.0 mg/kg of weight) for 14 (14-day RMA group) or 21 consecutive days (21-day RMA group). RMA was injected locally into the buccal mucosa of the left first maxillary molar twice daily starting 3 days before placement of the 10-gf Ni-Ti closed coil spring. Tooth movement distance was analysed using micro-computed tomography. The effects on surrounding alveolar bone were evaluated by measuring the ratio of bone surface area to tissue surface area with haematoxylin-eosin-stained sections and counting the number of osteoclasts in periodontal tissue with TRAP-stained sections. Blood tests were performed and bone volume and trabecular separation at the tibial neck were measured to analyse systemic side effects. RESULTS: Local RMA injection inhibited tooth movement by 40.6 per cent, promoted alveolar bone volume maintenance by 37.4 per cent, and inhibited osteoclast activity around the tooth root at 21 days by 40.8 per cent. Systemic effects on osteoclasts or osteoblasts were not observed. CONCLUSION: Local injection of RMA enabled control of tooth movement without systemic side effects in a mouse model.

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.

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.

DOCK8 mutation diagnosed using whole-exome sequencing of the dried blood spot-derived DNA: a case report of an Iraqi girl diagnosed in Japan.

BACKGROUND: Dedicator of cytokinesis 8 (DOCK8) deficiency (MIM #243700) is a rare disease, leads to a combined primary immunodeficiency (PID), and accounts for the autosomal recessive-hyper immunoglobulin E syndrome (AR-HIES). DOCK8 deficiency status characterizes by recurrent infections, atopy, and risk of cancer. Lymphoproliferative disease complicating PID, is difficult to diagnose. Our aim is to present a rare case of PID, and to the best of our knowledge, she is the first case of DOCK8 deficiency from Iraq. The genetic diagnosis was carried out in Japan using dried blood spot-based DNA transfer and whole-exome sequencing. CASE PRESENTATION: An 11-year-old Iraqi girl, of double first-cousin-parents, had a history of severe eczema, food allergy, and repeated infections. She presented with a jaw mass, bilateral cervical and axillary lymphadenopathy, and immunoglobulin (Ig) assays of 20, 3.3 and 1.7-fold above maximum normal level for age of IgE, IgA and IgG, respectively, along with a low IgM, eosinophilia and lymphopenia. Based on the jaw mass biopsy, non-Hodgkin lymphoma was suggested in Iraq, whereas histopathological re-evaluation in Japan revealed the diagnosis of a polyclonal reactive proliferation spectrum of lymphoproliferative disorders/plasmacytic hyperplasia, complicating PID. Whole-exome sequencing supported the diagnosis of PID by identifying a homozygous DOCK8 mutation with previously reported pathogenicity (NM_203447:c.3332delT, p.Phe1113Leufs*2), that may be attributed to consanguinity. CONCLUSIONS: International collaboration using an effective DNA transportation technique and next-generation sequencing was the key to pinpoint the diagnosis of DOCK8 deficiency. Our case asserted that careful pathogenetic evaluation, in an advanced setting, was crucial for ruling out the neoplastic process. Pediatricians in areas with a high prevalence of consanguinity marriage should have a high index of suspicion of DOCK8 deficiency in patients with recalcitrant eczema, and frequent respiratory and skin infectious episodes.

Effect of tablets containing lactoferrin and lactoperoxidase on gingival health in adults: A randomized, double-blind, placebo-controlled clinical trial.

OBJECTIVE: To evaluate the effect of tablets containing lactoferrin (LF) and lactoperoxidase (LPO) on gingival health and oral health-related quality of life in healthy adults. BACKGROUND: Lactoferrin and LPO are host defense factors found in saliva that may contribute to oral health. MATERIALS AND METHODS: One hundred and fifty adults were randomly assigned to the administration of high-dose tablets (LF 60 mg/d, LPO 7.8 mg/d), low-dose tablets (LF 20 mg/d, LPO 2.6 mg/d), or placebo tablets for 12 weeks. The gingival index (GI) and plaque index (PlI) were measured at baseline and after 12 weeks. Oral health-related quality of life was assessed by the Oral Health Impact Profile (OHIP) at baseline and at 4, 8, and 12 weeks. RESULTS: One hundred and nine healthy subjects were included in the efficacy analysis. In the high-dose group, the GI was significantly reduced after 12 weeks of treatment, and the reduction in GI in the high-dose group was significant compared with the placebo group. In both the high-dose group and the low-dose group, PlI showed a significant decrease at 12 weeks compared with baseline. The total OHIP score was significantly reduced at 12 weeks in the high-dose group. In addition, the OHIP functional limitation subscale displayed significant improvement in the high-dose groups compared with the placebo group at 12 weeks. No adverse reactions or serious adverse events related to the test tablets were observed in any of participants during the study, and the incidence of adverse events unrelated to the tablets did not differ significantly among the groups. CONCLUSION: These results suggest that intake of tablets containing LF (60 mg/d) and LPO (7.8 mg/d) can potentially improve gingival inflammation and oral health-related quality of life in healthy adults.

Increasing Risk of Disturbed Root Development in Permanent Teeth in Childhood Cancer Survivors Undergoing Cancer Treatment at Older Age.

BACKGROUND: Structural anomalies of teeth are observed at high rates in childhood cancer survivors (CCS). Several therapeutic exposures have been shown to be associated with dental developmental disturbances. This study was conducted to analyze the risk factors for dental developmental abnormality (DDA) and investigate the association between age at the time of cancer treatment and DDA in CCS. PATIENTS AND METHODS: Fifty-six CCS were enrolled. Orthopantomography and dental examination were performed in all the patients. We evaluated the prevalence of DDA and analyzed the risk factors for each type of DDA. RESULTS: DDAs were observed in 46.4% of CCS, including hypodontia in 9 (16.1%), abnormal roots in nine (16.1%), enamel defects/hypoplasia in 6 (10.7%), and microdontia in 12 (21.4%) patients. The number of patients with abnormal roots was significantly higher in the group treated with stem cell transplantation or at an age older than 4 years. We observed that the formation period of abnormal teeth coincided with the treatment period in the majority of CCS with DDA. CONCLUSIONS: Particularly regarding the root abnormality, treatment at elder age may be a risk factor for root developmental disturbances. Risk evaluation, appropriate follow-up, and early detection of dental issues are required for all CCS.

Success rate and strength of osseointegration of immediately loaded UV-photofunctionalized implants in a rat model.

STATEMENT OF PROBLEM: Despite its clinical benefits, the immediate loading protocol might have a higher risk of implant failure than the regular protocol. Ultraviolet (UV) photofunctionalization is a novel surface enhancement technique for dental implants. However, the effect of photofunctionalization under loading conditions is unclear. PURPOSE: The purpose of this animal study was to evaluate the effect of photofunctionalization on the biomechanical quality and strength of osseointegration under loaded conditions in a rat model. MATERIAL AND METHODS: Untreated and photofunctionalized, acid-etched titanium implants were placed into rat femurs. The implants were immediately loaded with 0.46 N of constant lateral force. The implant positions were evaluated after 2 weeks of healing. The strength of osseointegration was evaluated by measuring the bone-implant interfacial breakdown point during biomechanical push-in testing. RESULTS: Photofunctionalization induced hydrophilic surfaces on the implants. Osseointegration was successful in 28.6% of untreated implants and 100% of photofunctionalized implants. The strength of osseointegration in successful implants was 2.4 times higher in photofunctionalized implants than in untreated implants. The degree of tilt of untreated implants toward the origin of force was twice that of photofunctionalized implants. CONCLUSIONS: Within the limit of an animal model, photofunctionalization significantly increased the success of osseointegration and prevented implant tilt. Even for the implants that underwent successful osseointegration, the strength of osseointegration was significantly higher for photofunctionalized implants than for untreated implants. Further experiments are warranted to determine the effectiveness of photofunctionalization on immediately loaded dental implants.

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 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.

Stabilization of tooth movement by administration of reveromycin A to osteoprotegerin-deficient knockout mice.

INTRODUCTION: In this study, mechanical stress in the form of tooth movement was applied to osteoprotegerin-deficient knockout mice, which served as an animal model for juvenile Paget's disease. To compare and evaluate bone turnover and response of the surrounding bony tissue, we administered reveromycin A. We also investigated the ability of reveromycin A to control osteoclastic activity in juvenile Paget's disease. METHODS: Eight-week-old male osteoprotegerin-deficient knockout and wild-type mice were injected with reveromycin A (15 mg/kg of body weight) intraperitoneally twice daily. An elastic module was inserted interproximally between the maxillary left first and second molars. RESULTS: Administration of reveromycin A to osteoprotegerin-deficient knockout mice reduced tooth movement distances, increased bone volumes at the interradicular septum, decreased osteoclast counts, and reduced serum alkaline phosphatase and tartrate resistant acid phosphatase. Reveromycin A administration also caused a temporal shift in peak Runx2 staining in osteoprotegerin-deficient knockout mice so that the overall staining time course was similar to that observed for wild-type mice. CONCLUSIONS: Reveromycin A administration in osteoprotegerin-deficient knockout mice inhibited bone resorption and normalized bone formation. As a result, normal bone turnover was obtained.

Bisphosphonate inhibits bone turnover in OPG(-/-) mice via a depressive effect on both osteoclasts and osteoblasts.

Osteoclast differentiation and functioning are strictly controlled by RANKL expressed on osteoblast membrane surfaces, but whether osteoclasts exert control over osteoblasts remains unclear. In the present study, we examined the effect of an osteoclast inhibitor, a bisphosphonate (BP), on the response of maxillary bone to mechanical stress in a high-turnover osteoporosis model (OPG(-/-) mice, a model of juvenile Paget disease). Mechanical stress was induced by use of orthodontic elastics to move the maxillary first molar. BP was administered once per day beginning 5 days before elastic insertion. Relative to wild type (WT), in the OPG(-/-) mice tooth movement distance was greater, resorption of the interradicular septum occurred to a greater extent, the osteoclast count was higher, and serum alkaline phosphatase (ALP) was higher. However, administration of BP to OPG(-/-) mice reduced tooth movement distance, increased bone volume at the interradicular septum, decreased the osteoclast count, and reduced serum ALP. BP administration also caused a temporal shift in peak Runx2 staining in OPG(-/-) mice, such that the overall staining time course was similar to that observed for WT mice. We conclude that BP administration not only inhibited osteoclast activity in OPG(-/-) mice but also systemically and locally inhibited osteoblast activity. It is possible that osteoclasts are able to exert some negative control over osteoblasts.

Reveromycin A Administration Prevents Alveolar Bone Loss in Osteoprotegerin Knockout Mice with Periodontal Disease.

Chronic periodontal disease is characterized by alveolar bone loss and inflammatory changes. Reveromycin A (RMA) was recently developed and is a unique agent for inhibiting osteoclast activity. This study analysed the effects of RMA in an experimental mouse model of periodontitis involving osteoprotegerin (OPG)-knockout mice, specifically, whether it could control osteoclasts and reduce inflammation in periodontal tissue. We examined wild-type (WT) and OPG knockout mice (OPG KO) ligated with wire around contact points on the left first and second molars. RMA was administered twice a day to half of the mice. Using micro-computed tomography, we measured the volume of alveolar bone loss between the first and second molars, and also performed histological analysis. The OPG KO RMA+ group had significantly decreased osteoclast counts, alveolar bone loss, attachment loss, and inflammatory cytokine expression 8 weeks after ligation. Thus, RMA may reduce alveolar bone loss and inflamed periodontal tissues in patients with periodontitis.

[Clinical availability of the herbal medicine, SYOUSAIKOTOU, as a gargling agent for prevention and treatment of chemotherapy-induced stomatitis].

The stomatitis accompanying chemotherapy reduces a patient's QOL. Many reports have suggested that some kinds of gargling agents for oral mucositis shorten the duration and severity of symptoms. This study tested the prevention and efficacy against stomatitis of a herbal medicine (Syousaikotou) as a gargling agent for patients receiving chemotherapy. Compared to gargling with providone-iodine and amphotericin B, the Syousaikotou gargle showed a significantly decreased incidence of stomatitis, and a painkilling effect. Stomatitis occurred in about 17.4% among 23 chemotherapy cycles with the Syousaikotou gargle, against about 40.8% among 71 chemotherapy cycles without the Syousaikotou gargle. Among the patients suffering stomatitis pain after 22 chemotherapy cycles, the painkilling effect was seen to be 76.2%, and continues for about 2 hours. Critical side effects were not seen, but in 4 cases there were complaints about foul smells, such as oil and grass smells. Syousaikotou gargle was considered to be one of the useful methods against the stomatitis prevention and sharp pain mitigation from the chemotherapy.

Effects of a chewing gum containing phosphoryl oligosaccharides of calcium (POs-Ca) and fluoride on remineralization and crystallization of enamel subsurface lesions in situ.

OBJECTIVES: Manufacturers are adding fluoride (F) to calcium-containing chewing gums to further promote enamel remineralization. The aim of this study was to assess the effect of a chewing gum containing phosphoryl oligosaccharides of calcium (POs-Ca) and fluoride on remineralization of enamel subsurface lesions, in a double-blind, randomized controlled in situ trial. METHODS: Thirty-six volunteer subjects wore removable buccal appliances with three different insets of bovine enamel with subsurface demineralized lesions. For 14 days the subjects chewed one of the three chewing gums (placebo, POs-Ca, POs-Ca+F), three times a day. After each treatment period, the insets were removed from the appliance, embedded, sectioned, polished and then subjected to laboratory tests; mineral level was determined by transverse microradiography (TMR; n=36), and hydroxyapatite (HAp) crystallites were assessed by synchrotron radiation wide-angle X-ray diffraction (WAXRD; n=13). Data were analysed by t-test or Wilcoxon rank-sum test with Bonferroni corrections at 0.05 significance level. RESULTS: Chewing POs-Ca and POs-Ca+F gums resulted in 21.9±10.6 and 26.3±9.4 (mean±SD) percentage mineral recovery, which was significantly higher than that of placebo gum (15.0±11.4) (p<0.05). Chewing POs-Ca+F gum resulted in 24.9±5.4 (mean±SD) percentage HAp crystallites recovery, which was significantly higher compared to POs-Ca (16.0±4.1%) or placebo (11.1±4.8%) gums (p<0.05). CONCLUSIONS: Addition of POs-Ca to the chewing gum resulted in significant remineralization of enamel subsurface lesions. Although POs-Ca+F gum was not superior in TMR recovery rate when compared with POs-Ca gum, WAXRD results highlighted the importance of fluoride ion bioavailability in the formation of HAp crystallites in enamel subsurface lesions in situ (NCT01377493).