Layer Of New Bone Research Articles

Retention of fish-like odontode overgrowth in Permian tetrapod dentition supports outside-in theory of tooth origins.

Teeth are often thought of as structures that line the margins of the mouth; however, tooth-like structures called odontodes are commonly found on the dermal bones of many Palaeozoic vertebrates including early jawless fishes. 'Odontode' is a generalized term for all tooth-like dentine structures that have homologous tissues and development. This definition includes true teeth and the odontodes of early 'fishes', which have been recently examined to gain new insights into the still unresolved origin of teeth. Two leading hypotheses are frequently referenced in this debate: the 'outside-in' hypothesis, which posits that dermal odontodes evolutionarily migrate into the oral cavity, and the 'inside-out' hypothesis, which posits that teeth originated in the oropharyngeal cavity and then moved outwards into the oral cavity. Here, we show that, unlike the well-known one-to-one replacement patterns of marginal dentition, the palatal dentition of the early Permian tetrapods, including the dissorophoid amphibian Cacops and the early reptile Captorhinus, is overgrown by a new layer of bone to which the newest teeth are then attached. This same overgrowth pattern has been well documented in dermal and oral odontodes (i.e. teeth) of early fishes. We propose that this pattern represents the primitive condition for vertebrates and may even predate the origin of jaws. Therefore, this pattern crosses the fish-tetrapod transition, and the retention of this ancestral pattern in the palatal dentition of early terrestrial tetrapods provides strong support for the 'outside-in' hypothesis of tooth origins.

Novel osteoconductive β-tricalcium phosphate/poly(L-lactide-co-e-caprolactone) scaffold for bone regeneration: a study in a rabbit calvarial defect.

The advantages of synthetic bone graft substitutes over autogenous bone grafts include abundant graft volume, lack of complications related to the graft harvesting, and shorter operation and recovery times for the patient. We studied a new synthetic supercritical CO2 -processed porous composite scaffold of β-tricalcium phosphate and poly(L-lactide-co-caprolactone) copolymer as a bone graft substitute in a rabbit calvarial defect. Bilateral 12 mm diameter critical size calvarial defects were successfully created in 18 rabbits. The right defect was filled with a scaffold moistened with bone marrow aspirate, and the other was an empty control. The material was assessed for applicability during surgery. The follow-up times were 4, 12, and 24 weeks. Radiographic and micro-CT studies and histopathological analysis were used to evaluate new bone formation, tissue ingrowth, and biocompatibility. The scaffold was easy to shape and handle during the surgery, and the bone-scaffold contact was tight when visually evaluated after the implantation. The material showed good biocompatibility and its porosity enabled rapid invasion of vasculature and full thickness mesenchymal tissue ingrowth already at four weeks. By 24 weeks, full thickness bone ingrowth within the scaffold and along the dura was generally seen. In contrast, the empty defect had only a thin layer of new bone at 24 weeks. The radiodensity of the material was similar to the density of the intact bone. In conclusion, the new porous scaffold material, composed of microgranular β-TCP bound into the polymer matrix, proved to be a promising osteoconductive bone graft substitute with excellent handling properties.

Influence of biphasic calcium phosphate surfaces coated with Enamel Matrix Derivative on vertical bone growth in an extra-oral rabbit model.

The aim of this study was to investigate the ability of Enamel Matrix Derivative (EMD) on vertical bone regeneration around dental implants placed in an extra-oral rabbit model. A total of 30 Straumann BL implants were partially embedded in transverse orientation into the posterior mandibles of 15 rabbits. Macro-structuring BiPhasicCaPST (BCPT1), micro-structuring BiPhasicCaPST (BCPT2), and deproteinized bovine bone mineral (DBBM) were placed around the implant and covered with a scaffold stabilizing "umbrella." EMD was incorporated within the scaffold for test sites, but not control sites. Histological analysis was performed on retrieved specimens after 10weeks of healing to assess new bone formation. All treatment groups displayed new supracrestal bone formation as determined by histomorphometric measurements, with mean values of new bone height ranging between 0.62 and 1.13mm. Histological analysis revealed a higher mean bone formation (%) around the test sites where EMD (34.7, 95%CI: 27.1-39.4) was released from the scaffold, whereas the control group without EMD release (26.4, 95%CI: 16.3-31.9) (P=0.069). The mean fBIC (%) in the BCPT2 group increased by the addition of EMD relative to no EMD (67.2, 95%CI: 48.6-84.1) and (54.7, 95%CI: 32.3-68.9), respectively). The BCPT2/EMD and DBBM/EMD interventions showed the greatest mean bone density (BA/TA), respectively, (12.8, 95%CI: 8.9-36.5) and (11.2, 6.3-16.4) in ROI 1. Values in ROI 2 were, similarly, (24.9, 95%CI: 17.2-31.7) and (27.7, 19.2-35.3). BA/TA in ROI 2 differences between the BCPT2 groups with and without EMD was statistically significant (P=0.026), as well as the DBBM groups with and without EMD (P=0.038). A layer of new bone was formed in both test and controls. The release of EMD from BCPT2 and DBBM adjacent to a bone-level implant with an SLActive surface and scaffold retention umbrella consistently regenerated the greater fBIC and bone density along the length of the implant.

Biomineralization stimulated peri-titanium implants prepared by selective laser melting

Titanium implants prepared by selective laser melting (SLM), a method of additive manufacturing, were subjected to implantation in beagle dogs for two and four weeks. Argon ion beam-polished cross sections of the implants after in vivo tests were characterized by scanning electron microscope (SEM) to evaluate the bone–implant interface and the early peri-implant biomineralization with sufficiently improved resolution. Two bone mineralization mechanisms were disclosed. As early as two weeks after implantation, a layer of new bone was found to form directly on the implant surface and bone in-growth was also observed. Osseointegration was found to establish partly at the tip of the implants. After healing for four weeks it was found that osseointegration was established around the entire tip of the implants, whereas only partly at the third thread region of the implants. The experimental evidences observed reveal that an inherent highly porous surface of the titanium implants generated by selective laser melting is favorable for new bone apposition.

Alendronate-eluting polyglucose-lignol composite (POGLICO)

Background and purpose — Due to the known drawbacks of metal implants, new biomaterials for internal fracture fixation are attracting increasing interest, among them poly(lactic-co-glucolic) acids (PLGAs) and the recently developed silk-tenoin derived materials (STDMs). In accordance with the new philosophy of bio-derived biomaterials (BIODERIBIOs), I describe a novel innovative technology for use in fracture fixation.Patients and methods — Screws (2 mm dia.) were manufactured from cylindrical bars of polyglucose-lignol composite (POGLICO) in the form of birch toothpicks from the hospital canteen, dip-coated with alendronate (1 mg/mL, n = 6) or saline (n = 6), and inserted in the proximal tibias of rats for 4 weeks. Fixation was evaluated by mechanical pullout testing. POGLICO nails were inserted in the contralateral tibia for microCT and histology.Results — All POGLICO implants remained fixed in the bone (p < 0.001) with a mean pullout force of 37 (SD 5.5) N. MicroCT showed that the control nails were surrounded by a thin layer of new bone, while all bisphosphonate-treated implants were surrounded by a thick layer of cancellous bone. Bisphosphonates more than doubled the bone density around the nails (p = 0.004).Interpretation — POGLICO is biocompatible, remains in situ, and appears to provide a higher resistance to pullout forces than bulk silk protein. The material is light, strong, and bio-derived. BIODERIBIO-POGLICO can be sterilized by autoclaving, and has a porous surface that can serve for slow release of drugs applied by simple dip-coating, as demonstrated by the effect of the alendronate treatment. As the raw material for the screws is readily available from the toothpick industry, I believe that the possibilities for commercial development of the material for fracture fixation are promising.

Influence of bone affinity on the skeletal distribution of fluorescently labeled bisphosphonates in vivo

Bisphosphonates are widely used antiresorptive drugs that bind to calcium. It has become evident that these drugs have differing affinities for bone mineral; however, it is unclear whether such differences affect their distribution on mineral surfaces. In this study, fluorescent conjugates of risedronate, and its lower-affinity analogues deoxy-risedronate and 3-PEHPC, were used to compare the localization of compounds with differing mineral affinities in vivo. Binding to dentine in vitro confirmed differences in mineral binding between compounds, which was influenced predominantly by the characteristics of the parent compound but also by the choice of fluorescent tag. In growing rats, all compounds preferentially bound to forming endocortical as opposed to resorbing periosteal surfaces in cortical bone, 1 day after administration. At resorbing surfaces, lower-affinity compounds showed preferential binding to resorption lacunae, whereas the highest-affinity compound showed more uniform labeling. At forming surfaces, penetration into the mineralizing osteoid was found to inversely correlate with mineral affinity. These differences in distribution at resorbing and forming surfaces were not observed at quiescent surfaces. Lower-affinity compounds also showed a relatively higher degree of labeling of osteocyte lacunar walls and labeled lacunae deeper within cortical bone, indicating increased penetration of the osteocyte canalicular network. Similar differences in mineralizing surface and osteocyte network penetration between high- and low-affinity compounds were evident 7 days after administration, with fluorescent conjugates at forming surfaces buried under a new layer of bone. Fluorescent compounds were incorporated into these areas of newly formed bone, indicating that "recycling" had occurred, albeit at very low levels. Taken together, these findings indicate that the bone mineral affinity of bisphosphonates is likely to influence their distribution within the skeleton.

Implant system for guiding a new layer of bone. Computed microtomography and histomorphometric analysis in the rabbit mandible

To prove the concept that an implant system with osteoconductive surface characteristics and an osteoinductive scaffold material has the capacity to guide vertical supracrestal bone growth in a rabbit mandible onlay model. Thirteen adult white New Zealand rabbits each received custom-designed dental implants. All implants had sandblasted, acid-etched (SLA) surfaces, with the coronal aspect (3 mm) of each implant was left outside the lateral aspect of posterior mandibular bone, but covered by periosteum, muscle, subcutaneous tissue, and skin. Bone formation around implants placed adjacent to osteoinductive demineralized bone matrix (DBM) scaffolds were compared with contralateral implants without scaffolds in six rabbits using micro-CT imaging. Bone formation around implants with scaffolds from seven additional rabbits was measured using both micro-CT imaging and quantitative histology. At 8 weeks, new supracrestal bone was seen adjacent to all implants placed with DBM and two implants without DBM. The mean supracrestal bone heights achieved for implants with and without DBM scaffolds as measured by micro-CT was 2.1+/-0.9 and 0.8+/-0.9 mm, respectively (P=0.008). Histomorphometric analysis illustrated that supracrestal bone-to-implant contact for implants with DBM scaffolds was 58.1+/-14% and that mean supracrestal bone height was 2.4+/-0.6 mm. Successful implant-guided supracrestal osteogenesis has been demonstrated in a rabbit model with the combined use of osteoconductive implant surfaces, an osteoinductive scaffold, and a device that prevents soft tissue downgrowth and provides scaffold stabilization.

Growth of new bone guided by implants in a murine calvarial model

New methods to increase vertical bone growth are needed to permit dental implant placement in patients with low alveolar ridge height after extended periods of tooth loss. While ectopic rodent models are typically used to evaluate new osteogenic implant surface coatings, a more relevant intramembraneous rodent model was needed to address the particular clinical need to grow a new layer of bone above an existing layer of bone. In this study we report on a novel murine calvaria model in which successful vertical bone growth around miniaturized dental implants was achieved when using non-glycosylated bone morphogenetic protein-2 (ng/rhBMP-2). Twenty CD-1 mice received two Ti implants each consisting of a Ti ring implant stabilized by a Ti screw into the occipital calvarial bone. Four groups were evaluated: control Ti, Ti + 20 μg ng/rhBMP-2, hydroxyapatite (HA)-coated Ti, and HA + 20 μg ng/rhBMP-2. The mice were sacrificed 21 days following implant placement. MicroCT analysis showed no new bone formation around the untreated Ti or the HA-coated implants, but demonstrated new bone growth in every dimension around and above the Ti + ng/rhBMP-2 and the HA + ng/rhBMP-2 treated implants. Histopathologic analysis showed that a thin fibrous capsule covered the untreated Ti implants. Limited bone-to-implant contact (BIC) was observed for the HA-coated implants, while in contrast both ng/rhBMP-2 treated groups exhibited extensive new supracalvarial woven bone that covered the implant and merged with the calvarial plate. Histomorphometrically, supracalvarial bone heights and bone widths and BIC were not statistically different from one another for the two ng/rhBMP-2 treated groups. However, the total supracalvarial bone surface area was significantly greater ( p < 0.05) for the Ti + ng/rhBMP-2 implants (7.2 mm 2) than the HA + ng/rhBMP-2 (4.0 mm 2) treated implants. The bone density within 1 mm around the implant was also significantly greater ( p < 0.05) for the Ti + ng/rhBMP-2 implants (9.9%) than the HA + ng/rhBMP-2 (4.0%) implants, indicating that HA coatings may not be required for sustained release when non-glycosylated BMP-2 is used. This new murine model is capable of discriminating between various bone augmentation strategies and may represent a clinically more relevant model for alveolar bone augmentation than the commonly used ectopic muscle pouch or long bone models.

In vivo bone response to 3D periodic hydroxyapatite scaffolds assembled by direct ink writing

The in vivo bone response of 3D periodic hydroxyapatite (HA) scaffolds is investigated. Two groups of HA scaffolds (11 mm diameter x 3.5 mm thick) are fabricated by direct-write assembly of a concentrated HA ink. The scaffolds consist of cylindrical rods periodically arranged into four quadrants with varying separation distances between rods. In the first group, HA rods (250 microm in diameter) are patterned to create pore channels, whose areal dimensions are 250 x 250 microm(2) in quadrant 1, 250 x 500 microm(2) in quadrants 2 and 4, and 500 x 500 microm(2) in quadrant 3. In the second group, HA rods (400 microm in diameter) are patterned to create pore channels, whose areal dimensions of 500 x 500 microm(2) in quadrant 1, 500 x 750 microm(2) in quadrants 2 and 4, and 750 x 750 microm(2) in quadrant 3. Each group of scaffolds is partially densified by sintering at 1200 degrees C prior to being implanted bilaterally in trephine defects of skeletally mature New Zealand White rabbits. Their tissue response is evaluated at 8 and 16 weeks using micro-computed tomography, histology, and scanning electron microscopy. New trabecular bone is conducted rapidly and efficiently across substantial distances within these patterned 3D HA scaffolds. Our observations suggest that HA rods are first coated with a layer of new bone followed by subsequent scaffold infilling via outward and inward radial growth of the coated regions. Direct-write assembly of 3D periodic scaffolds composed of micro-porous HA rods arrayed to produce macro-pores that are size-matched to trabecular bone may represent an optimal strategy for bone repair and replacement structures.

The preparation, structure evaluation and preliminary application of biomimetic biphasic calcium phosphate scaffold

To fabricate biomimetic biphasic calcium phosphate BCP ceramic scaffolds using three-dimensional (3D) gel-lamination technology and evaluated their structure with 3D parameters and related method. Series two-dimensional images of femoral head's specimen of dogs were obtained by micro-computed tomography (Micro-CT). According to these images, porous biomimetic biphasic calcium phosphate (BCP) ceramic scaffolds with oriented trabecular structure were fabricated by three-dimensional (3D) gel-lamination technology. And then, the three-dimensional structure of the scaffolds were reconstructed by computer according to Micro-CT images of these scaffolds and evaluated by three-dimensional parameters. These parameters included bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular spacing (Tb.Sp) and structure model index (SMI). The biomechanical properties and biocompatibility of these scaffolds were also evaluated in the study. Six scaffolds, which were combined with BMCs (bone mesenchymal cells, BMCs), were planted into the bone defect of six dogs' femoral head respectively. There was no significant difference between trabecular samples and BCP scaffolds in BV/TV, Tb.Th, Tb.N, and Tb.Pf (P > 0.05). The trabecular system of the scaffold, which had some orientation, represented plate-like model. With a micro-porous porosity of 62%, the average compressive modulus and ultimate strength along the axis of the scaffolds reached (464.0 +/- 36.0) MPa and (5.6 +/- 0.8) MPa respectively. The results of animal test indicated that the trabeculae of these scaffolds were covered by a layer of new bone after 10 weeks of operation. Porous BCP scaffolds have been produced with oriented microarchitectural features designed to facilitate vascular invasion and cellular attachment and with initial mechanical properties comparable to those of trabecular bone.

Bone formation at titanium porous oxide (TiUnite™) oral implants in type IV bone

Several oral implant design advances have been suggested to overcome poor bone quality, an impediment for successful implant treatment. A novel titanium porous oxide (TPO) surface has been shown to offer favorable results in several settings. The objective of this study was to evaluate the local bone formation and osseointegration at TPO-modified implants in type IV bone. Three TPO surface-modified implants (TiUnite) were installed into the edentulated posterior maxilla in each of 8 Cynomolgus monkeys. The animals were injected with fluorescent bone labels at 2, 3, 4 and 16 weeks post-surgery and were euthanized at week 16 when block biopsies were collected for histologic analysis. The predominant observation of the TPO implant surface was a thin layer of new bone covering most of the implant threads. Mean (+/-SE) bone-implant contact for the whole study group was 74.1 +/- 4.8%. There was a significant variability in bone-implant contact between animals (P = 0.0003) and between sites of the same animal (P < 0.0001). The variance in bone-implant contact was 30% larger among sites of the same animal than between different animals (187.5 vs. 144.8, respectively). There was a small but significant difference in bone density immediately outside, compared to within the threaded area of the implants (37.1 +/- 3.2% vs. 32.1 +/- 3.2%, P < 0.0001). Bone density outside the implant threads was significantly correlated (beta = 0.682, P < 0.0001) with the bone density within the threaded area. Bone density within the threaded area was significantly correlated (beta = 0.493, P = 0.0002) with bone-implant contact, whereas bone density outside the implant threads did not have a significant effect (beta = 0.232, P = 0.1). The results suggest that the TPO surface possesses a considerable osteoconductive potential promoting a high level of implant osseointegration in type IV bone in the posterior maxilla.

Augmentation of tendon-bone healing by the use of calcium-phosphate cement.

The healing of a hamstring graft to bone is the weak link in the reconstruction of a cruciate ligament using this donor material. We therefore investigated the augmentation of healing at the tendon-bone interface using calcium-phosphate cement (CPC). We performed semitendinosus autograft reconstructions of the anterior cruciate ligament on both knees of 22 New Zealand white rabbits. The interface between the grafted tendon and the bone tunnel for one knee was filled with CPC. Six rabbits were killed at the end of the first and second post-operative weeks in order to evaluate the biomechanical changes. Two rabbits were then killed sequentially at the end of weeks 1, 3, 6, 12 and 24 after operation and tissue removed for serial histological observation. Histological examination showed that the use of CPC produced early, diffuse and massive bone ingrowth. By contrast, in the non-CPC group of rabbits only a thin layer of new bone was seen. Mechanical pull-out testing at one week showed that the mean maximal tensile strength was 6.505 +/- 1.333 N for the CPC group and 2.048 +/- 0.950 N for the non-CPC group. At two weeks the values were 11.491 +/- 2.865 N and 5.452 +/- 3.955 N, respectively. Our findings indicate that CPC is a potentially promising material in clinical practice as regards its ability to reinforce the fixation of the tendon attachment to bone and to augment the overall effectiveness of tendon healing to bone.

Stimulation of craniofacial and intramedullary bone formation by negatively charged beads

To test for their osteogenic stimulating capacity, charged beads were implanted into cranial or mandibular defects, used as an onlay on the nasal bone surface, or injected into femoral medullary cavities of young adult rats. One month later, negatively charged beads were found to have stimulated extensive bone formation resulting in closure of craniofacial defects, a new layer of bone on the nasal bone surface, and a bead-bone lattice within marrow cavities of long bones. Positively charged beads were nonosteogenic, but elicited a pronounced fibroblastic response in the craniofacial skeleton. Positively charged beads were found associated with multinucleated giant cells at all implantation sites. Uncharged beads failed to elicit formation of new bone and were associated with connective tissue that was less cellular and less organized than was seen with positively charged beads. It was concluded that beads that have been chemically treated to confer either a negative or positive surface charge, when placed in contact with bone, evoke osteogenesis or formation of dense connective tissue, the response depending on the surface charge of the bead. The mechanism(s) by which the charged beads foster the osteogenic or fibroblastic response is not clear. The use, however, of alloplastic materials with charged surfaces in repair and augmentation of bone, and in wound repair, warrants further investigation.

The effect of intermittent cortisone administration in the rabbit: induction of cycles of resorption and deposition of bone.

The intermittent administration of cortisone in both the young and the mature rabbit is associated with appositional bone growth on the periosteal surfaces of the cranium, premaxilla and middle of the shaft of the femur; each new layer of bone is separated from the next by a darkly haematoxylin-staining "reversal" line. The internal architecture of the bone also changes in consequence of the repeated waves of resorption and deposition of bone round vascular spaces. Cartilaginous growth at the epiphysis in the young rabbit is also affected. The long columns of metaphysial cartilage are replaced by a layer of new bone which partly seals the epiphysial cartilage from the marrow spaces.

家鶏下顎骨の発生学的研究

The development of the mandible was studied in the hatching embryo, the chicken and the adult domestic fowl, and the results obtained are summarized as follows. 1. The constituent bones of the mandible include Dental, Supraangular, Splenial, Gonial and Angular. The last mentioned bone, however, lacks independence because it fuses with the Articular (at the Processus angularis internus) in the adulthood of the animal. Of the anlages of the coverning bones, those of the Supraangular and Angular appear at seven and a half days after hatching, the Dental at 8 and a half days, the Splenial at 9 days and the Angular at 10 days after hatching. Completion of the covering hones : These bones are constructed into their mature forms within 20 days after hatching, but the time needed for total completion including that of the Articular bone is 3 months after birth. If, further, the formation of the mandibular canal and the fusion of each constituent bone are therein included, the time needed for is one year after birth. 2. The formation and completion of the Articular bone : The Articular bone is formed and completed by cooperation of enchondrous and intramembranous ossifications by 3 months after birth. 3. Formation of the mandibular canal : This is made primarily by intramembranous ossification at the posterior part of the Meckel's cartilage which is then gradually fused with the Supraangular. 4. Formation of the Angular bone : The anlage of the bone develops on the 10th embryonic day at the inner side of that portion corresponding to the root of Processus angularis internus, and by the 11th day a thin layer of new bone is deposited along the process, later to increase the bulk gradually. On the 15the embryonic day cartilage cells occur on the posterior dorsal part of the bone facing the process, and on the 20th day a part of the bone is already fused with the process. By 2 months after hatching the whole constituent bones become completely integrated so that no further discrimination is feasible. 5. Fusion of the constituent bones and how it occurs : Until 6 months after birth the constituent bones are packed together dorsoventrally and in-and outwardly, but by the 12 th month after birth they are almost fused. The fusion takes place as metaplasmatic bone formation especially with regard to the Gonial, Supraangular and the posterior part of the Angular fitting into the concavity on the ventral side of the Articular. 6. Later course of the Meckel's cartilage : After the formation of the Articular bone, the remaining cartilage gradually shorten its antero-posterior length by intramembranous ossification arising from its front and rear peripheries.