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Background/purpose: While there are numerous reports on surgical techniques and materials for bone grafting, limited methods are available to enhance the body's inherent capacity to heal bones. Here we investigated microRNA-199a (miR-199a), a molecular that promotes osteoblast differentiation and bone healing. Materials and methods: To construct a miR-199a delivery complex, miR-199a-5p mimics were coated with mesoporous silica nanoparticles (MSNs) following modified with polyethyleneimine (PEI) and peptide WEAKLAKALAKALAKHLAKALAKALKACEA (KALA) to obtain 199a-5p-loaded MSN-PEI-KALA. Nanoparticle complexes are assessed for particle size and zeta potential using transmission electron microscopy and dynamic light scattering. Then MC3T3-E1 cells are exposed to MSN_miR-199a-5p @PEI-KALA. The impact of MSN_miR-199a-5p@PEI-KALA at varying concentrations on cell viability is assessed using Cell Counting Kit-8. Cell uptake and distribution were analyzed by double fluorescent staining with fluorescein amidite-labeled MSN_miR-199a@PEI-KALA and lysosome labeling. On day 7 after osteogenic induction, alkaline phosphatase (ALP) staining was conducted. Results: The findings indicated that the nanoparticle complexes encapsulating PEI and peptide exhibited an augmentation in both particle size and zeta potential. At a dosage of 10 μg/mL, MSN_miR-199a@PEI-KALA displayed the lowest cytotoxicity compared to the control group. MC3T3-E1 cells treated with MSN_miR-199a-5p@PEI-KALA exhibited intensified ALP staining and elevated mRNA expression levels of ALP, runt-related transcription factor 2, and osteopontin, suggesting the involvement of miR-199a-5p-loaded MSN-PEI-KALA in osteogenic differentiation. Conclusion: The successful construction of the delivering complex MSN_miR-199a@PEI-KALA in present research highlights the promise of this biomaterial carrier for the application of miRNAs in treating bone defects.

Background/purpose: Porcine collagen is widely used in regenerative therapies to generate membranes for bone augmentation. However, porcine or bovine gelatin or collagen is often not appropriate for patients with creed and religious beliefs or for allergic reasons. In this study, we evaluated the potential of fish gelatin to generate membranes. Materials and methods: Fish gelatin and hydroxyapatite (HAp) were used at three different ratios (2:0, 2:1, 2:1.5, and 2:2) to prepare gelatin-hydroxyapatite (G-HAp) membranes via freeze-drying and heat-crosslinking. The surface morphology and cell attachment of G-HAp membranes were observed using scanning electron microscopy and confocal laser microscopy. G-HAp membrane was placed at the bottom of a well plate, and MC3T3-E1 cells were seeded on it. Cell viability and cytotoxicity were tested after 1 and 3 days of culture. Alkaline phosphatase (ALP) and alizarin red staining was performed at 10 and 21 days, respectively. Results: Viability of cells on G-HAp membrane with the gelatin:HAp ratio of 2:1.5 was significantly higher than that on membranes with other gelatin:HAp ratios. ALP and alizarin red staining showed that ALP-positive areas and calcium deposition were the highest on G-HAp membrane with the gelatin:HAp ratio of 2:1. These membranes showed negligible cytotoxicity. Conclusion: Fish-derived G-HAp membranes have the potential to promote osteogenic differentiation of MC3T3-E1 cells with negligible cytotoxicity.

Abstract Background Salivary alpha-amylase (sAA) activity level is thought to be an indicator of mental stress. However, the relationship between sAA activity levels and mental stress in patients during dental implant treatment has not been studied. In the present study, we aimed to examine the correlation between sAA activity levels and changes in patients’ vital signs during dental implant surgery. Results Levels of sAA activity were higher after surgery when compared to before-surgery measurements. A significant positive correlation was found between sAA activity and heart rate (HR) (r s =0.434, p=0.007) as well as a positive correlation with oxygen level (r s =0.392, p=0.016). Conclusion Levels of sAA activity tended to increase after the surgical procedures, as did patients’ stress levels. SpO2 and sAA activity levels were inversely correlated. There was a positive significant correlation between HR and sAA activity, though there was no correlation between blood pressure and sAA activity levels. Salivary alpha-amylase may be a valuable indicator of stress and anxiety in dental patients undergoing dental implant surgery.

Abstract Background The application of dental implants is often restricted by bone volume. In such cases, bone grafts are required, although bone graft materials have some disadvantages. Therefore, other effective approaches are needed. Our previous study showed that the autologous micrograft, a dissociated cell suspension made out of palatal connective tissue grafts, promoted bone-marrow cell proliferation and differentiation under osteogenic conditions. In this study, we aimed to evaluate the effects of dissociated soft-tissue suspensions relevant to bone regeneration in animal model. Material and methods Twelve-week-old male Wistar rats were used in the study. Defects were created in rat calvaria, and were filled with hydrogel containing either dissociated soft-tissue suspension (test) or sucrose (control). The new bone formation was evaluated at 1 and 2 weeks after surgery (n = 16) by radiological and histological analysis. Results The conducted radiological analysis showed that the new bone volume was significantly greater in the dissociated soft-tissue suspension group. This finding was further confirmed by the conducted histological analysis. Conclusions The dissociated mucosa tissue suspension enhanced bone regeneration in vivo; thus, it is a promising potential method to aid the successful application for bone augmentation in the implant practice.

Abstract Background To evaluate the effects of electrolysis on cleaning the contaminated healing abutment surface and to detect the optimal condition for cleaning the contaminated healing abutment. Methods Ninety healing abutments removed from patients were placed in 1% sodium dodecyl sulfate solution and randomly divided for electrolysis with 7.5% sodium bicarbonate in the following three different apparatuses (N = 30): two stainless steel electrodes (group I), a copper electrode and a carbon electrode (group II), and two carbon electrodes (group III). The samples were placed on cathode or anode with different electric current (0.5, 1, and 1.5 A) under constant 10 V for 5 min. Electrolyte pH before and after electrolysis were measured. Then, the samples were stained with phloxine B and photographed. The proportion of stained areas was calculated. The surface was examined with a scanning electron microscope (SEM) and energy-dispersive X-ray spectroscopy (EDS). Results Electrolyte pH decreased after electrolysis at 1 A and 1.5 A in group I and II. Applying cathode at 1 A in group III, the amount of residual contamination was the lowest in all the conditions examined in the present study. SEM images revealed that applying cathode at 1.5 A in group I induced a rough surface from the smooth surface before the treatment. EDS analysis confirmed that the surfaces treated on cathode at 1 A in group III revealed no signs of organic contamination. Conclusion Electrolysis of using carbon as electrodes, placing the contaminated healing abutments on cathode, and applying the electric current of 1 A at constant 10 V in 7.5% sodium bicarbonate could completely remove organic contaminants from the surfaces. This optimized electrochemical cleaning method seems to be well worth investigation for the clinical management of peri-implant infections.

Abstract Background We performed maxillary sinus floor augmentation using the crestal approach and beta-tricalcium phosphate (β-TCP) and a long-term evaluation using cone-beam computed tomography (CBCT). Methods Twenty-three patients (30 implants) underwent sinus floor augmentation using the osteotome technique. Subjects underwent CBCT imaging before surgery, immediately after surgery, and at follow-up (≥ 3 years after surgery). We measured the changes in height of the augmented sinus floor (SL), the augmented bone above apex of the implant (BH), and the implant length that projected into the sinus (IL). Results The mean SL decreased from 6.54 ± 1.51 (immediately after surgery) to 3.11 ± 1.35 mm (follow-up). The mean BH decreased from 3.17 ± 0.97 to − 0.25 ± 1.19 mm; the maxillary sinus floor in many implants was near the apex at follow-up. The SL at follow-up showed a strong correlation with the IL (p = 0.0057). Conclusions Osteotome sinus floor elevation with beta-tricalcium phosphate was clinically effective. Cone-beam computed tomography analysis revealed that ≥ 3 years after surgery, the maxillary sinus floor was near the apex of the implant.

Abstract Background Previous studies have shown that porous composite blocks containing uncalcined hydroxyapatite (u-HA; 70 wt%) with a scaffold of poly-DL-lactide (PDLLA, 30 wt%) are biodegradable, encourage appropriate bone formation, and are suitable for use as a bone substitute in vertical ridge augmentation. The present study aimed to accelerate osteogenesis in vertical ridge formation by adding types 1 and 3 collagen to the u-HA/PDLLA blocks and assessing the effect. Material and methods The bone substitute in the present study comprised porous composite blocks of u-HA (70 wt%) with a PDLLA (27–29 wt%) scaffold and enriched with types 1 and 3 collagen (1.7 ~ 3.4 wt%). The control blocks were composed of u-HA (70 wt%) and PDLLA (30 wt%). The materials were formed into 8-mm diameter, 2-mm high discs and implanted onto the cranial bones of six rabbits. The animals were sacrificed 4 weeks after implantation, and histological and histomorphometrical analyses were performed to quantitatively evaluate newly formed bone. Results New bone formation occurred with both block types, showing direct contact with the original bone. Mean ± standard deviation bone formation was significantly greater in the experimental blocks (25.6% ± 4.8%) than in the control blocks (17.0% ± 4.7%). Conclusions Histological and histomorphometrical observations indicated that new bone was formed with both block types. The u-HA/PDLLA block with types 1 and 3 collagen is a more promising candidate for vertical ridge augmentation than the u-HA/PDLLA alone block.

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Despite numerous treatment methods, there is no gold standard for the treatment of peri-implantitis—an infectious peri-implant disease. Here, we examined selenium nanoparticles (SeNPs) at a wide range of concentrations to investigate their cytotoxicity, regulation of osteoblastic differentiation, and assessed the antibacterial effect against Porphyromonas gingivalis. SeNPs (mean size: 70 nm; shape: near-spherical; concentration: 0–2048 ppm) were tested against the MC3T3-E1 osteoblast precursor cell line and P. gingivalis red complex pathogen. Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) analysis was used to evaluate the bone morphogenetic protein 2 (BMP-2) signaling pathway. SeNPs at concentrations of 2–16 ppm showed no obvious cytotoxicity and promoted good mineralization and calcification. SeNPs at concentrations 64 ppm and below influenced gene expression promoting osteoblastic differentiation, whereas at high concentrations inhibited the expression of Runt-related transcription factor 2 (Runx2). The growth of P. gingivalis was significantly inhibited at SeNP concentrations of more than 4 ppm. SeNPs at low concentrations promoted osteoblastic differentiation while strongly inhibiting peri-implantitis pathogen growth. This study represents one of the few in vitro assessments of SeNPs against a red complex pathogen and the regulatory effect on osteoblastic differentiation. The findings demonstrate SeNPs could potentially be used for future application on implant coating.

Peri-implantitis and periodontitis are both polymicrobial diseases induced by subgingival plaque accumulation, with some differing clinical features. Studies on the microbial and gene transcription activity of peri-implantitis microbiota are limited. This study aimed to verify the hypothesis that disease-specific microbial and gene transcription activity lead to disease-specific clinical features, using an integrated metagenomic, metatranscriptomic, and network analysis. Metagenomic data in peri-implantitis and periodontitis were obtained from the same 21 subjects and metatranscriptomic data from 12 subjects were obtained from a database. The microbial co-occurrence network based on metagenomic analysis had more diverse species taxa and correlations than the network based on the metatranscriptomic analysis. Solobacterium moorei and Prevotella denticola had high activity and were core species taxa specific to peri-implantitis in the co-occurrence network. Moreover, the activity of plasmin receptor/glyceraldehyde-3-phosphate dehydrogenase genes was higher in peri-implantitis. These activity differences may increase complexity in the peri-implantitis microbiome and distinguish clinical symptoms of the two diseases. These findings should help in exploring a novel biomarker that assist in the diagnosis and preventive treatment design of peri-implantitis.

Background and Objectives: Oral implant restorations are an excellent treatment option for edentulous patients; however, periodontopathogenic bacteria have been found in the microgaps between implant−abutment junctions. Implant designs to limit the microgaps have been extensively studied. However, studies have shown microgaps continue to exist, allowing for the leakage of bacteria into the implant system. Screw access hole materials are used to fill the access hole void. The use of materials with beneficial properties could provide bacterial leakage prevention. The aim of this study was to examine the surface free energy, cytotoxicity, and bacterial adhesion of selected screw access hole materials such as cotton, polytetrafluoroethylene (PTFE) tape, paraffin wax−polyolefin thermoplastic (PF), paraffin wax (Wax), gutta-percha (GP), and caviton EX (CE). Materials and Methods: A sessile drop test was performed to observe the contact angle and calculate the surface free energy of each material in order to determine the level of hydrophobicity. Cytotoxicity was examined in a mouse gingival epithelial cell line for day 1 and day 3. Bacterial adhesion was tested with Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans. Results: PTFE, PF, and wax presented low surface free energies of 19.34, 23.041, and 24.883 mN.m-1, respectively. No cytotoxicity was observed, except for GP and CE. Concurrently, the bacterial adhesion was also the lowest in PTFE and PF. Conclusions: Within the limits of this study, PTFE and PF showed an excellent biocompatibility with few bacterial adhesions. These materials could be potential screw access hole materials in clinical settings.

(1) Background: When dental implants are placed at the esthetic zone, facial bone fenestration might be expected. This study aimed to evaluate the biomechanical effect of bone augmentation around implants with facial bone fenestration defects using the finite element method. (2) Methods: An anterior maxillary region model with facial concavity was constructed with a threaded implant inserted following the root direction, resulting in apical threads exposure to represent the fenestration model. Several bone coverage levels were simulated by gradually shifting the deepest concavity point buccally, mimicking bone augmentation surgeries with different bone fill results. Oblique forces were applied, and analysis was performed. (3) Results: Peak compressive stress magnitude and distribution varied according to the level of exposure and facial concavity depth. The fenestration model demonstrated a slightly lower peak peri-implant bone stress, smaller implant displacement, and smaller bone volume with strain levels above 200 µ strain. A gradual increase in compressive stress, implant displacement, and bone volume exhibited strain level above 200 µ strain was observed with the increased bone fill level of the facial bone fenestration. (4) Conclusions: Exposure of implants apical threads at the maxillary anterior region does not significantly affect the peri-implant stress and strain results. However, increasing the buccolingual width and eliminating the buccal concavity might increase the peri-implant bone volume exhibited favorable loading levels.

Purpose: To evaluate the performance of sealing materials used in the screw-access holes of screw-retained implant final superstructures in vivo and in vitro. Materials and Methods: Twenty-one screw-access holes in the final superstructures were randomly divided into three groups (each group, n = 7). Following disinfection and isolation, all access holes were initially filled with sterilised cotton pellets of the same weight. Depending on the group, the access holes were finally sealed with either provisional composite restorations (group A), self-curing resin for provisional sealing (group B), or acrylic resin (group C). After one month of the functional period, the inner cotton pellets were collected as bacterial reservoirs. Results: Total aerobic bacteria and total gram-negative anaerobic bacteria were measured after bacterial culture for 48 h and 72 h, respectively. In vitro evaluation of porosity using scanning electron microscopy (SEM) was also performed. Samples from superstructures sealed with provisional composite restorations showed fewer bacteria and less porosity than samples from superstructures sealed with self-curing resin for provisional sealing and acrylic resin. In this study, provisional composite restorations showed the best sealing properties. Provisional composite restorations may prevent bacterial invasion of the access holes of the final superstructures. Conclusion: In this study, provisional composite restorations showed the best sealing properties. Provisional composite restorations may prevent bacterial invasion of the access holes of the final superstructures. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the positive effects of a CPC-, GK2-, and TXA-based (CPC/GK2/TXA) mouthwash after implant placement. Materials and Methods: Twenty patients (n = 20) who underwent posterior implant-placement surgery were randomly and evenly allocated to the study or the placebo group. After the mouthwash was used 3x/day for 7 to 10 days postoperatively, sutures were analysed by counting the colony-forming units (CFU) for total aerobes, total G [-] anaerobes, total enterobacteria and total H. influenzae, followed by Real-Time PCR of bacterial-specific DNAs of A. actinomycetemcomitans, P. gingivalis, T. forsythia, T. denticola, P. intermedia, P. micra, F. nucleatum, C. rectus, and E. corrodens. In vitro resistance of P. gingivalis, S. aureus, and P. aeruginosa was analysed. The compatibility of the mouthwash with Straumann SLA implant surfaces was evaluated by scanning electron microscopy (SEM). Results: Sixteen patients (n = 16) completed the trial. A statistically significantly greater number of CFU was found in the placebo group for almost all species, especially for total G [-] anaerobes. No statistically significant in vitro resistance was found for P. gingivalis, S. aureus, and P. aeruginosa. SEM revealed no surface alteration after exposure to the mouthwash. Conclusion: The use of a CPC/GK2/TXA mouthwash inhibited propagation of the bacteria extracted from the postsurgical sutures after implant placement. [ABSTRACT FROM AUTHOR]

Objective: This study evaluated the cleaning potential of a protein-denaturing agent with or without anionic detergent by monitoring the residual contamination on healing abutments used for dental implant treatment. Method and materials: Forty contaminated healing abutments removed from patients were randomized and immediately treated with differing cleaning methods; either Method A (presoaking in 1% sodium dodecyl sulfate followed by ultrasonication with 4 mol/L guanidine hydrochloride), or Method B (soaking in distilled water followed by ultrasonication with 4 mol/L guanidine hydrochloride) was used. Samples were stained with phloxine B and photographed using a light microscope. The proportion of stained and contaminated areas on each healing abutment was then calculated using Image J. The surface was examined with a scanning electron microscope and energydispersive x-ray spectroscopy. Results: The percentages of contaminated surfaces of the screwdriver engagement region, upper body, and lower body for methods A and B were 50% and 38%, 10% and 80%, and 38% and 18%, respectively. There was a statistically significant difference (engagement region [P < .001], upper body [P = .043], and lower body [P = .017]; Mann-Whitney) regarding the residually stained areas between the two cleaning methods. No surface alterations were seen by scanning electron microscopy. Energy-dispersive x-ray spectroscopy confirmed that the cleaned surfaces of the healing abutments revealed no signs of organic contamination. Conclusion: Although the combination of a strong denaturing agent and detergent effectively cleaned contaminated healing abutments, perfect cleaning was not always possible, indicating that the reuse of healing abutments in different patients is not recommended. [ABSTRACT FROM AUTHOR]

This study evaluated the quantity and quality of newly formed vertical bone induced by sputtered hydroxyapatite-coated titanium implants compared with sandblasted acid-etched implants after dura mater elevation. Hydroxyapatite-coated and non-coated implants (n = 20/group) were used and divided equally into two groups. All implants were randomly placed into rabbit calvarial bone (four implants for each animal) emerging from the inferior cortical layer, displacing the dura mater 3 mm below the original bone. Animals were sacrificed at 4 (n = 5) and 8 (n = 5) weeks post-surgery. Vertical bone height and area were analyzed histologically and radiographically below the original bone. Vertical bone formation was observed in both groups. At 4 and 8 weeks, vertical bone height reached a significantly higher level in the hydroxyapatite compared with the non-coated group ( p

Purpose: The purpose of this in vitro study was to examine the effect of dissociated soft tissue on bone marrow cell proliferation and differentiation under osteogenic conditions. Materials and Methods: Rat bone marrow cells were cultured to assess the stimulation of cell proliferation and differentiation. Harvested palatal mucosa was dissociated using a device (Rigenera, Human Brain Wave), and the dissociated soft tissue was cultured with rat bone marrow cells. Cell proliferation, differentiation, and mineralized nodule formation were assessed after 2 or 5 days of culturing. Bone marrow cell proliferation was assessed by quantifying the absorbance of a water-soluble tetrazolium salt using a cell proliferation assay kit. Bone marrow cell differentiation was assessed by alkaline phosphatase staining and real-time polymerase chain reaction. Mineralized nodule formation was assessed by Alizarin red staining. Results: At day 2, cell proliferation, osteoblast-specific gene expression, and mineralized nodule formation were significantly higher in the experimental group than in the control group. Alkaline phosphatase staining was also higher in the experimental group on day 2. Mineralized nodule formation area and osteoblast-specific gene expression were also statistically higher in the experimental group on day 5. Conclusion: This study demonstrates that dissociated soft tissue elevates bone marrow cell proliferation and differentiation under osteogenic conditions. [ABSTRACT FROM AUTHOR]

Periosteum has been demonstrated to contain mesenchymal progenitor cells differentiating to osteoblasts, and both bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) may play important roles in cell-based approaches to bone regeneration. The purpose of this study was to evaluate the feasibility and efficacy of BMP-2 and/or VEGF on periosteal cell differentiation to osteoblasts in vitro and ectopic bone formation in vivo. Human periosteum-derived cells were transfected with BMP-2, VEGF, BMP-2 + VEGF, or vehicle as a control by non-viral gene transfer and then cultured and implanted to nude mice intramuscularly. Real-time polymerase chain reaction analysis of the culture revealed that transgenes for BMP-2 and BMP-2 + VEGF induced more mRNA expression of alkaline phosphatase, collagen type I, and osteocalcin than VEGF and vehicle treatments; additionally, alizarin red S staining, alkaline phosphatase staining, and alkaline phosphatase activity were significantly higher in the BMP-2 + VEGF transgene than in the other versions. After implantation, ectopic bone was observed at 4 weeks and greatly increased at 8 weeks in all groups. In particular, the combination of BMP-2 and VEGF formed significantly more bone at 4 weeks, and VEGF transfection resulted in more blood vessels relative to the conditions without VEGF. Thus, VEGF might enhance BMP2-induced bone formation through modulation of angiogenesis. Keywords:: tissue engineering, periosteal cell, bone morphogenetic protein-2 (BMP-2), vascular endothelial growth factor (VEGF), gene transfer

In vivo gene transfer with plasmid vector has been applied experimentally and clinically; however, the low level of gene transfer efficiency with plasmid vector is a problem. We speculated that the combination of calcium phosphate precipitate (CaP) and plasmid vector could solve this problem because CaP stabilizes plasmid DNA. In the present study, we used a plasmid exression vector encoding enhanced green fluorescent protein and combined the vector with CaP. Then, this combination was mixed with bovine type I atelocollagen. After incubating this mixture in phosphate-buffered saline, the amount of the plasmid DNA in the supernatant was low when the plasmid DNA was combined with CaP. Furthermore, the plasmid DNA, which was combined with CaP, was stable in DNase digestion in vitro. The plasmid vector with or without CaP, together with the atelocollagen, was transplanted subcutaneously or injected in the bone marrow of the femurs of rats. Then, the fluorescence was observed under a confocal laser scanning microscope and the fluorescence intensity in the tissue homogenates was measured. In these animal experiments, the fluorescence was extensive when the plasmid DNA was combined with CaP. These results indicate that our formula, collagen/CaP/DNA, appeared efficient for in vivo gene transfer. Keywords:: calcium phosphate, plasmid, gene transfer, atelocollagen, enhanced green fluorescent protein

Simvastatin, a cholesterol synthesis inhibitor, enhances BMP2 expression in osteoblasts. The purpose of the present study was to examine whether simvastatin stimulates bone regeneration when combined with calcium sulfate as a carrier. Critical-sized bone defects in rat calvaria were treated with calcium sulfate or with combination of 1 mg simvastatin and calcium sulfate. In the combination group, although the least amount of bone formation with intense soft tissue inflammation was observed at 2 and 4 weeks, remarkable bone formation was evident at 8 weeks. Conclusively, the combination of simvastatin and calcium sulfate stimulated bone regeneration in spite of the inflammatory response. Keywords:: simvastatin, calcium sulfate, bone formation

Adipose tissue-derived stromal cells, termed ASCs, play an important role in regenerative applications. They resemble mesenchymal stem cells owing to their inexhaustibility, general differentiation potential, and plasticity and display a series of cell-specific and cluster-of-differentiation (CD) marker profiles similar to those of other somatic stem cells. Variations in phenotypes or differentiation are intimately associated with CD markers. The purpose of our study was to exhibit distinct populations of ASCs with differing characteristics for osteogenic differentiation. The primary cell batch of murine-derived ASCs was extracted from subcutaneous adipose tissue and the cells were sorted for the expression of the surface protein molecules CD90 and CD105 using flow cytometry. Each cell population sorted for CD90 and CD105 was analyzed for osteogenic potency after cell culture. The results suggested that ASCs exhibit distinct populations with differing characteristics for osteogenic differentiation: unsorted ASCs stimulated comparable mineralized nodule formation as bone marrow stromal cells (BMSCs) in osteogenic medium and viral transfection for BMP2 accelerated the formation of mineralized nodules in CD90 and/or CD105 positive ASCs with observation of decrease in CD105 expression after 14 days. Future studies assessing different immunophenotypes of ASCs should be undertaken to develop cell-based tissue engineering.

Guided bone regeneration is a unique osteogenesis technique that requires a barrier membrane under periosteum to create space for bone regeneration. However, creating sizeable spaces is clinically not commonly feasible. A titanium plate and a thin silicone membrane were surgically layered on each calvaria of eight rabbits. Then, the periphery of the silicone membrane was fixed by a plastic ring to the underlying bone using titanium micro screws. After 1 week, a 5-mm-length titanium screw was used to elevate the titanium plate, which in turn elevated the silicone membrane together with overlying soft tissue in a rate of 1 mm/day for 5 days to create a secluded space. Animals were killed at 2 months (n = 4, group 1) and 4 months (n = 4, group 2) after the elevation. Histological and microradiographical analyses demonstrated creation of an amount of de novo bone formation (68.2 ± 22 mm 3 in group 1 and 70.3 ± 14 mm 3 in group 2) in the sizeable created spaces (207.1 ± 31 mm 3 in group 1 and 202 ± 21 mm 3 in group 2) without exposure of the device. This novel osteogenesis technique, “expansible guided bone regeneration,” created a substantial in vivo incubator without applying growth factors or osteoprogenitor cells. Creating a growing space over the secluded surface allowed the development of normal biological healing process occurring on the bone surface into a regenerative process, generating bone outside the genetically determined skeletal bone. This technique is a new tissue engineering approach stimulating endogenous tissue repair without applying cells or factors exogenously.

Donor organ transplantation is currently an essential therapeutic approach to the replacement of a dysfunctional organ as a result of disease, injury or aging in vivo. Recent progress in the area of regenerative therapy has the potential to lead to bioengineered mature organ replacement in the future. In this proof of concept study, we here report a further development in this regard in which a bioengineered tooth unit comprising mature tooth, periodontal ligament and alveolar bone, was successfully transplanted into a properly-sized bony hole in the alveolar bone through bone integration by recipient bone remodeling in a murine transplantation model system. The bioengineered tooth unit restored enough the alveolar bone in a vertical direction into an extensive bone defect of murine lower jaw. Engrafted bioengineered tooth displayed physiological tooth functions such as mastication, periodontal ligament function for bone remodeling and responsiveness to noxious stimulations. This study thus represents a substantial advance and demonstrates the real potential for bioengineered mature organ replacement as a next generation regenerative therapy.

Purpose: Among potential cell-based therapies, adipose-derived stem cells (ASCs) have been proposed as a promising source of stem cells for tissue regeneration. Although many recent clinical trials have investigated the use of adipose tissue or ASCs in transplantation, analysis of the microstructures of outgrowing macrosized spheroids (macrospheroids) or three-dimensional coculture of ASC spheroids and monocyte/macrophage lineages has not been performed. The aim of this study was to analyze the microstructures of murine-derived ASC macrospheroids and the growth and osteogenic potential of these macrospheroids in a three-dimensional environment and after calcification induction by coculture with monocytes. Materials and Methods: The histologic structures of murine-derived ASC macrospheroids and the expression of marker genes for multipotency within these macrospheroids were analyzed by hematoxylin and eosin staining and in situ hybridization. ASC macrospheroid microstructures were observed by transmission electron microscopy, and cell proliferation in the spheroids was analyzed. Additionally, the growth and osteogenic potential of these macrospheroids were assessed in two-dimensional and three-dimensional environments and after calcification induction by coculture with monocytes. Results: The expression of Oct3/4, Nanog, and Sox2 was detected even in the deep zone of spheroids, although higher expression was observed at the surface. Cell proliferation was detected within the spheroid centers. Observation of spheroid microstructure revealed extracellular matrix production within the spheroid architecture. Transplantation of a spheroid on the hydroxyapatite disc resulted in three-dimensional cell growth, filling the disc. Coculture of the spheroids with monocytes led to the formation of many osteoclast-like, multinucleated cells, and calcification was observed after 3 weeks of coculture. Conclusion: ASC spheroids exhibited high capacity for dynamic three-dimensional growth and osteogenic differentiation. Furthermore, ASC spheroids promoted monocyte differentiation into osteoclast-like cells, which may enhance the osteogenic potential of ASC spheroids. [ABSTRACT FROM AUTHOR]

Members of the bone morphogenetic protein (BMP) family have diverse physiological roles. For instance, BMP-2 stimulates osteogenesis, while BMP-12 induces the formation of tendon/ligament-like tissues. Here, we designed a study to determine whether BMP-12 has bone and/or cartilage regeneration abilities similar to those of BMP-2. We implanted plasmid vectors encoding either BMP-2 or BMP-12 in rats with femur defects, and monitored the bone healing process for 8-weeks. The BMP-12 transgene induced prominent fibrogenesis by 2 weeks, with bone substitution occurring by 8 weeks. BMP-2, however, was associated predominantly with osteogenesis throughout the 8 week period. Thus, we conclude that BMP-12 does not function similarly to BMP-2 during bone healing. Further work is needed to better understand the mechanisms by which it stimulates bony growths to replace the connective tissues formed during the first stages of bone healing.