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Biomedical subjects

L P Garetto

Publications and source records attributed to L P Garetto.

At least 19 recordsLinked to original sources

The use of small titanium screws for orthodontic anchorage.

The use of conventional dental implants for orthodontic anchorage is limited by their large size. The purpose of this study was to quantify the histomorphometric properties of the bone-implant interface to analyze the use of small titanium screws as an orthodontic anchorage and to establish an adequate healing period. Overall, successful rigid osseous fixation was achieved by 97% of the 96 implants placed in 8 dogs and 100% of the elastomeric chain-loaded implants. All of the loaded implants remained integrated. Mandibular implants had significantly higher bone-implant contact than maxillary implants. Within each arch, the significant histomorphometric indices noted for the "three-week unloaded" healing group were: increased labeling incidence, higher woven-to-lamellar-bone ratio, and increased osseous contact. Analysis of these data indicates that small titanium screws were able to function as rigid osseous anchorage against orthodontic load for 3 months with a minimal (under 3 weeks) healing period.

Analysis of Variance↗

Animal housing influences the response of bone to spaceflight in juvenile rats.

The rat has been used extensively as an animal model to study the effects of spaceflight on bone metabolism. The results of these studies have been inconsistent. On some missions, bone formation at the periosteal bone surface of weight-bearing bones is impaired and on others it is not, suggesting that experimental conditions may be an important determinant of bone responsiveness to spaceflight. To determine whether animal housing can affect the response of bone to spaceflight, we studied young growing (juvenile) rats group housed in the animal enclosure module and singly housed in the research animal holding facility under otherwise identical flight conditions (Spacelab Life Science 1). Spaceflight reduced periosteal bone formation by 30% (P < 0.001) and bone mass by 7% in single-housed animals but had little or no effect on formation (-6%) or mass (-3%) in group-housed animals. Group housing reduced the response of bone to spaceflight by as much as 80%. The data suggest that housing can dramatically affect the skeletal response of juvenile rats to spaceflight. These observations explain many of the discrepancies in previous flight studies and emphasize the need to study more closely the effects of housing (physical-social interaction) on the response of bone to the weightlessness of spaceflight.

Animals↗

Implant-borne suture expansion in rabbits: a histomorphometric study of the supporting bone.

Osseointegrated implants have a large potential for diverse clinical applications, including support for sutural expansion and facial prostheses. The objectives of this study were to evaluate: (1) the histomorphometric response of thin cortical bone to implant placement and (2) whether loading of the bone surrounding these implants affects osseointegration as evaluated by histomorphometry. Eighteen New Zealand White rabbits had two titanium implants placed bilaterally in the anterior surface of their nasal bones. The rabbits were divided into an unloaded control group, one experimental group loaded at 1 Newton (N), and another loaded at 3 N. Fluorescent labels were used to mark areas of active bone formation. All rabbits were euthanized after 12 weeks of loading. Stereological point-hit and line-intercept methods were used to measure bone volume, direct bone-implant contact, new bone volume, and bone turnover rate in the bone surrounding the implants. All the implants remained stable during the loading period. A factorial ANOVA with repeated measures was used to compare the variables. The only significant difference among the three groups was a higher bone volume in the lateral coronal far region in the control group (p < 0. 05). Within all groups, bone volume (p < 0.002), turnover rate (p < 0.001), and percent of new bone (p < 0.05) were higher within 1 mm of the implant compared to 1-3 mm away. This may be due to the increased stress and strain in the bone adjacent to the implant. This study indicates that there are no detrimental effects of loading on osseointegration when implants placed in the thin facial cortices are used as anchors for sutural expansion.

Animals↗

Development of a fluorescent light technique for evaluating microdamage in bone subjected to fatigue loading.

A new method using fluorescent light microscopy has been developed to visualize and evaluate bone microdamage. We report the findings of two different experiments with a common aim of comparing the fluorescent light technique to the brightfield method for quantifying microdamage in bone. In Experiment 1, 36 canine femurs were tested in four-point cyclic bending until they had lost between 5 and 43% of their stiffness. The loaded portion of the bone was stained en bloc with basic fuchsin for the presence of damage. Standard point counting techniques were used to calculate fractional damaged area (Dm.Ar = Cr.Ar/B.Ar, mm2/mm2) under brightfield and fluorescent microscopy. In Experiment 2, bone microdamage adjacent to endosseous implants, subjected to fatigue loading (150,000 cycles, 2 Hz and 37 degrees C) ex vivo was examined. The bone around the implant was either allowed to heal (adapted specimen) for 12 weeks after placement in dog mid-femoral diaphyses prior to testing or was loaded immediately to simulate non-healed bone surrounding endosseous implants (non-adapted). Crack numerical density (Cr.Dn = Cr.N/B.Ar, #/mm2), crack surface density (Cr.S.Dn = Tt.Cr.Le/B.Ar, mm/mm2) and fractional damaged area were calculated separately by both techniques in the adapted and non-adapted specimens. In both Experiments 1 and 2, significantly more microdamage was detected by the fluorescent technique than by the brightfield method. Also, there was a trend towards higher intraobserver repeatability when using the fluorescent method. These results suggest that the brightfield technique underestimates microdamage accumulation and that the fluorescent technique better represents the actual amounts of microdamage present. The results demonstrate that the fluorescent method provides an accurate and precise approach for bone microdamage evaluation, and that it improves the prediction of stiffness loss from damage accumulation.

Animals↗

Microdamage adjacent to endosseous implants.

Intense remodeling occurs in lamellar bone adjacent to osseointegrated endosseous implants. The purpose of this study was to compare microdamage accumulation subsequent to ex vivo fatigue loading of bone that surrounds an endosseous implant, (a) immediately after placement (nonadapted bone) and (b) following a 12 week healing period after placement (adapted bone). We hypothesize that there is less microdamage in the more compliant adapted bone than in the older nonadapted bone. Nonthreaded titanium plasma sprayed (TPS)-coated endosseous implants were placed into dog mid-femoral diaphyses and allowed to heal for 12 weeks. Block sections of bone, each containing one implant, were cut anteroposteriorly, resulting in an implant containing lateral cortex, and a medial cortex that was used for testing the nonadapted specimens. Control specimens (n = 14 each for adapted and nonadapted) were loaded at 0 N. Experimental specimens (n = 13, adapted; n = 14, nonadapted) were loaded at 100 N in cantilever bending for 150,000 cycles at 2 Hz, at 37 degrees C on a Bionix 858 testing machine. Specimens were bulk stained with basic fuchsin and 120-140 microm sections were obtained. Crack numerical density (Cr.Dn = Cr.N/ B.Ar, #/mm2), crack surface density (Cr.S.Dn = Tt.Cr.Le/ B.Ar, mm/mm2), and percent damage area (Dm.Ar = Cr.Ar x 100/B.Ar, mm2/mm2) were measured at x 250. Statistically significant differences (p < 0.0001) were seen for Cr.Dn, Cr.S.Dn, and Dm.Ar on the compressed cortices suggesting that adapted bone near the implant accumulated significantly less microdamage than nonadapted bone. Also, the adapted nonloaded control specimens had approximately 20-fold less damage than the respective nonadapted specimens. This study suggests that the compliant adapted bone adjacent to endosseous implants is relatively resistant to fatigue loads. The high success rates of endosseous implants may be due to the presence of a rapidly remodeling region that maintains tissue compliance and limits microdamage initiation.

Animals↗

Fluoride treatment increased serum IGF-1, bone turnover, and bone mass, but not bone strength, in rabbits.

We hypothesized that fluoride partly acts by changing the levels of circulating calcium-regulating hormones and skeletal growth factors. The effects of oral fluoride on 24 female, Dutch-Belted, young adult rabbits were studied. The rabbits were divided into two study groups, one control and the other receiving about 16 mg fluoride/rabbit/day in their drinking water. After 6 months of fluoride dosing, all rabbits were euthanized and bone and blood samples were taken for analyses. Fluoride treatment increased serum and bone fluoride levels by over an order of magnitude (P < 0.001), but did not affect body weight or the following serum biochemical variables: urea, creatinine, phosphorus, total protein, albumin, bilirubin, SGOT, or total alkaline phosphatase. No skeletal fluorosis or osteomalacia was observed histologically, nor did fluoride affect serum PTH or Vitamin D metabolites (P > 0.4). BAP was increased 37% (P < 0.05) by fluoride; serum TRAP was increased 42% (P < 0.05); serum IGF-1 was increased 40% (P < 0.05). Fluoride increased the vertebral BV/TV by 35% (P < 0.05) and tibial ash weight by 10% (P < 0.05). However, the increases in bone mass and bone formation were not reflected in improved bone strength. Fluoride decreased bone strength by about 19% in the L5 vertebra (P < 0.01) and 25% in the femoral neck (P < 0. 05). X-ray diffraction showed altered mineral crystal thickness in fluoride-treated bones (P < 0.001), and there was a negative association between crystal width and fracture stress of the femur (P < 0.02). In conclusion, fluoride's effects on bone mass and bone turnover were not mediated by PTH. IGF-1 was increased by fluoride and was associated with increased bone turnover, but was not correlated with bone formation markers. High-dose fluoride treatment did not improve, but decreased, bone strength in rabbits, even in the absence of impaired mineralization.

Acid Phosphatase↗

Angiogenesis and osteogenesis in an orthopedically expanded suture.

The purpose of this study was to examine the angiogenic and the subsequent osteogenic responses during a 96-hour time-course after sutural expansion. Fifty rats were divided into: (1) a control group that received only angiogenic induction through injection of 5 ng/gm recombinant human endothelial cell growth factor (rhECGF); (2) an experimental group that received orthopedic expansion and rhECGF; (3) a sham group that received expansion and sodium chloride (NaCl) injection; and (4) a baseline group that received no expansion or injection. All rats were injected with 3H-thymidine (1.0 microCi/gm) 1 hour before death to label the DNA of S-phase cells. Demineralized sections (4 microm thick) were stained with hematoxylin and eosin. Angiogenesis and cell migration were analyzed with a previously established cell kinetics model. Analysis of variance was used to test the hypothesis that enhancement of angiogenesis stimulates reestablishment of osteogenic capability. Blood vessel number, area, and endothelial cell-labeled index significantly increased in experimental groups, but no difference was found between control and baseline groups. Labeled-pericyte index and activated pericyte numbers in the experimental group were also higher than in the sham groups. These results show that supplemental rhECGF enhances angiogenesis in expanded sutures but not in nonexpanded sutures. Data also suggest that pericytes are the source of osteoblasts in an orthopedically expanded suture.

Analysis of Variance↗

Sutural expansion using rigidly integrated endosseous implants: an experimental study in rabbits.

Rigidly integrated implants offer great promise for orthodontic and orthopedic anchorage in the oral and midfacial regions. Rigid anchorage can be used to control unwanted tooth movement, provide abutments in edentulous arches, and open the vertical dimension of occlusion. To evaluate the use of endosseous implants in the midface region, two flanged titanium implants were placed on either side of the midnasal suture of 18 New Zealand White rabbits. The rabbits were divided into an unloaded control and two experimental groups. One experimental group was loaded at 1 Newton (N) and the other at 3 N. All rabbits were euthanized after 12 weeks of loading. Stereologic point-hit and line-intercept methods were used to analyze microradiographic and multiple fluorochrome histology of the suture. All implants remained stable during the loading period. The distance between the implants increased significantly in the loaded groups compared with the control, and was significantly higher in the 3 N group than in the 1 N group. Percent bone volume was significantly decreased, while the percent suture volume tended to be increased in the loaded groups. Mineral apposition and bone formation rates at the sutural surfaces were increased in the loaded groups (P < 0.05), but did not differ between loaded groups. These results indicate that relatively low loads (1 or 3 N) applied to rigidly integrated endosseous implants across an unfused suture are satisfactory for achieving expansion under the conditions of this study. The 3 N load resulted in slightly more expansion, but did not affect the rate of bone formation at the suture.

Animals↗

Osteoporosis risk factors in female dental patients. A preliminary report.

Osteoporosis (OP) is a systemic condition that has dental implications. The purpose of this study was to compare OP risk among various dental specialty subpopulations at Indiana University School of Dentistry (IUSD). A survey was administered to 220 adult female dental patients assessing menstrual status and risk behaviors associated with low bone mass. The subjects' mean age was 48.2 +/- 1.1 years (mean +/- SEM). Overall, 38% of the surveyed patients exhibited high risk for OP. The orthodontic subpopulation (a dentate group with routine developmental malocclusions) was the youngest group and contained the lowest percentage at high-risk (6%). Conversely, the complete denture subpopulation was the oldest and contained the highest percentage of patients at high risk (53%). Postmenopausal women who had inadequate hormone replacement therapy exhibited a strong negative correlation for number of teeth retained with increasing years postmenopause (r = 0.6). Patients in the implant therapy group (many of which had adjunctive orthodontic care) had a mean age similar to the complete denture group, but a much lower risk for OP. This appears to be due to the extensive counseling these patients receive prior to treatment. It is concluded that risk factor analysis and patient counseling may be effective measures for reducing the osteoporosis risk of dental patients.

Adolescent↗

Histomorphometrical analysis of the bone-implant interface: comparison of microradiography and brightfield microscopy.

Section thickness has been shown to affect the histomorphometrical measurement of bone-implant contact when analysed under brightfield microscopy. This study investigated whether microradiography of the bone-implant interface eliminated the errors associated with thick section analysis. Seven implant containing sections were utilized. Microadiographs of the thick (approximately 100 microns) sections were taken and the sections were subsequently ground to thicknesses of 50 microns and 25 microns. Photomicrographs were taken of the microradiographs and of the sections at each thickness (100, 50 and 25 microns) under brightfield microscopy. The photomicrographs were analysed for direct bone-implant contact in the cortical passage region and along the total length of the implant. The effect of section thickness on multiple fluorochrome labelling in 10 rabbit femur specimens was also examined. Centre-to-centre interlabel distance was measured for each label pair at a thickness of 100 microns and then again after the sections were ground to 50 microns and 25 microns. The thick (100 microns) sections showed a significantly greater amount of bone-implant contact than either the thin sections or the microradiographs. There was no difference in direct bone-implant contact measured by microradiography or thin sections. However, the microradiographic analysis showed a much lower variability of the bone-implant contact than the sections evaluated under brightfield microscopy. In addition, they have the added benefit of providing information on bone mineral density. Centre-to-centre interlabel distance was not significantly different for any label pair owing to section thickness. Data from this study provides evidence that the use of microradiographs for histomorphometrical analysis of the bone-implant interface is superior to brightfield analysis of thin sections owing to the lower variability of microradiographical data and the ability to obtain bone mineral density measures. Additionally, given that interlabel distance was not significantly affected by section thickness, the use of 100 microns thick sections for analysis of fluorochrome labels in cortical bone is supported.

Analysis of Variance↗

Angiogenic induction and cell migration in an orthopaedically expanded maxillary suture in the rat.

The purpose was to examine the effect of an angiogenic factor on cell migration patterns and osteoblast histogenesis during the 96 h following orthopaedic expansion of the anterior maxillary suture. Fifty rats were divided into four groups: (1) a control group that received only angiogenic induction via injection of 5 ng/g body wt recombinant human endothelial-cell growth factor; (2) an experimental group that received orthopaedic expansion and angiogenic induction; (3) a sham group that received orthopaedic expansion and normal saline injection; and (4) a baseline group that received no expansion or injection. The experimental and sham groups were subdivided to conduct experiments over 1, 2, 3 or 4 days. The anterior portion of each maxilla was dissected free and demineralized. Sections (4 microns thick) were cut from every block and stained with Mayer's haematoxylin and eosin. Cell migration was analysed using a previously established cell-kinetics model. The osteoprogenitor cells were divided into four categories according to nuclear volume: A cells (40-79 microns3), B cells (80-119 microns3), C cells (120-169 microns3) and D cells (> or = 169 microns3 A' cells are the portion of the A cell population that responds to osteogenic stimulus. As previously defined in periodontal ligament, the reciprocal association of a decreasing number of less differentiated (A + A) cells and an increasing number of C + D cells, as a function of distance from the nearest major blood vessel, was consistently found in all groups. This suggests a vascularly oriented gradient of progressively more differentiated osteoprogenitor cells. Also, A + A' cells were predominately located within 20 microns of the nearest major blood vessel whereas the C + D cells were found at a distance > 30 microns from the nearest major blood vessel. These results suggest that the A'-->C shift occurs 20-30 microns from the nearest major blood vessel. In the angiogenic induction groups, the numbers of committed osteoprogenitors (A + A') were significantly higher than in the sham group at day 1. At day 3, the numbers of preosteoblasts (C + D) in angiogenic sutures were significantly higher than in the sham groups. This enhancement of preosteoblast population strongly suggests the possible role of activated pericytes in expanded sutures as a source of osteoprogenitor cells.

Analysis of Variance↗

Statistical analysis of differential lissajous EMG from normal occlusion and Class III malocclusion.

The method of differential lissajous electromyography (DL-EMG) was applied to investigate the relationship among the integrated EMG activity, timing, and coordination of the bilateral superficial anterior temporal and masseter muscle activities in normal occlusion and Class III malocclusion subjects. In both Class III malocclusion and normal occlusion subjects, the working side muscles showed a higher mean cumulative voltage (MCV) and mean maximum peak voltage (MMPV) compared with the balancing side. In addition, a higher MCV and MMPV of the working side masseter was observed in the normal occlusion group compared with that seen in the Class III group during both right and left side chewing (p < 0.01). Discriminant analysis applied to examine the distribution, the size and the shape of DL-EMG pattern further indicated a statistical difference between subject groups (p < 0.01). Finally, there was a significantly higher percentage of clockwise DL-EMG pattern-generation in the normal group compared with that seen for Class III subjects (p < 0.01). These data indicate that, compared with normal subjects, patients with a Class III malocclusion have a demonstrably abnormal masticatory muscle balance which is well characterized by the DL-EMG method.

Adolescent↗

Remodeling dynamics of bone supporting rigidly fixed titanium implants: a histomorphometric comparison in four species including humans.

The long-term maintenance of a rigid bone-implant interface (osseointegration) is the clinical goal of most dental implant systems, although the biological mechanism for retaining a foreign object in living bone is unclear. Little data are available on the physiological turnover (remodeling) of the supporting osseous tissue. The objective of this study was to histomorphometrically assess bone remodeling surrounding rigidly integrated titanium implants in multiple species. Implants, in place from 6 months to 5 years, were recovered from human, monkey, dog, and rabbit subjects. With the use of stereological point-hit and linear-intercept methods, indices of bone formation and resorption were determined. Remarkably similar patterns emerged among all investigated species. Repeated-measures ANOVA showed a 3 to 9 fold increase in remodeling within 1 mm of the bone-implant interface (P<0.001; data expressed as percent turnover / month, mean +/- SEM for n = 3-11). All morphometric indices (percent new bone, percent fluorochrome-labeled bone, percent resorption space) showed similar trends. These data suggest that the physiological mechanism for maintaining rigid osseous integration (osseointegration) is a sustained elevation of remodeling adjacent to the bone-implant interface.

Alveolar Process↗

Mechanical response to functional and therapeutic loading of a retromolar endosseous implant used for orthodontic anchorage to mesially translate mandibular molars.

Three finite element models were created to investigate the potential of rigid osseous fixation (osseointegration) for orthodontic anchorage: a mandible without an implant; a mandible with an implant; and a mandible and implant with a superimposed orthodontic load. Force was applied to different locations and the stresses were computed. The mechanical stress distributions adjacent to the implant were not affected by different biting forces, hence only one case needed to be analyzed. The stresses adjacent to the bone-implant interface changed drastically due to implantation, with major changes occurring on the buccal and mesiobuccal sides. A strong, concordant gradient for intraosseous stress and bone remodeling rate was observed that reflects a mismatch in the moduli of elasticity between the implant and the supporting bone. These results suggest important clinical implications. Osseointegration of symmetrically threaded titanium implants appears to be maintained by a sustained elevation of the mechanical stresses that continuously stimulate the bone remodeling activity within 1 mm of the implant surface. It is unlikely that a rigidly fixed (osseointegrated) implant will lose integration due to an orthodontic load superimposed on normal function.

Alveolar Process↗

A histomorphometric study on the healing of class III furcations utilizing bone labelling in beagle dogs.

The dynamics of bone turnover in the furcations of teeth treated with expanded polytetrafluoroethylene (ePTFE) membranes were evaluated using multiple fluorochrome labels in 6 male beagle dogs. Loss of attachment involving the furcation area was induced in the second, third, and fourth premolar teeth using silk ligatures. The resulting defects were treated with the use of mucoperiosteal flaps for access, debridement of the defects, and placement of ePTFE membranes covering the furcations of the second and fourth premolars (experimental teeth) while the third premolar received only debridement without membrane placement (control tooth). Five fluorochrome labels were administered intravenously at timed intervals to act as markers of the osseous response. Membranes were removed at 4 weeks and all animals were terminated at 12 weeks post-membrane placement. One side of the mandible was decalcified, sectioned at 7 microns, and stained with either hematoxylin and eosin or Gomori's tri-chrome. The opposite side provided non-decalcified tissue processed as 100 microns ground sections. Using fluorescent light and point-hit evaluation, tissue in the coronal half of each specimen was classified as either labelled bone, unlabelled bone, or resorption space. In addition, microradiographs were prepared of each ground section and specimens classified as either woven bone, old lamellar bone, or new lamellar bone. No significant differences in attachment levels, or level of junctional epithelium, were observed in decalcified sections although greater remodeling activity was noted in the experimental specimens. Comparison of ground sections revealed significant differences (P < 0.05) in all categories with both methods of evaluation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Bone physiology and metabolism in dental implantology: risk factors for osteoporosis and other metabolic bone diseases.

Placing a dental implant elicits a time-dependent bone response controlled by wound-healing factors (cytokines, bioelectrical signals), biomechanics (gravitational, functional, and therapeutic loads), and mineral metabolism (hormones, diet, excretion). The osseous response to an implant involves four physiological stages: (1) endosteal and periosteal callus formation; (2) compaction and remodeling of the callus; (3) remodeling (turnover) of the nonvital interface and adjacent bone; and (4) maturation (secondary mineralization) of new bone. Long-term maintenance of a rigid implant interface is related to continual bone remodeling. Common metabolic bone disorders affecting potential implant patients are osteopenia ("osteoporosis"), renal osteodystrophy, osteomalacia, and Paget's disease. The most prevalent problem is a long-term negative calcium balance leading to a compromise in bone strength. Symptomatic osteoporosis (usually wrist, hip, and/or spine fractures) affects 4 to 50 percent of the population depending on age, race, sex, endocrine status, and life-style. Postmenopausal white and Asian females present the greatest risk. The jaws of "osteoporotic" adults are variably affected because of the moderating influence of mechanical function. Management of metabolic bone disorders is an important consideration in diagnosis, treatment planning, and long-term monitoring of dental implants. Bone metabolic counseling, a natural extension of preventative dentistry, is an unexpected benefit readily appreciated by patients and their families.

Bone Diseases, Metabolic↗

Preosteoblast production in COSMOS 2044 rats: short-term recovery of osteogenic potential.

The influence of a 13.8-day spaceflight and approximately 8.5-11 h of recovery at 1 g on fibroblast-like osteoblast precursor cells was assessed in the periodontal ligament of rat maxillary first molars. Preosteoblasts (C + D cells), less differentiated progenitor cells (A + A' cells), and nonosteogenic fibroblast-like cells (B cells) were identified by nuclear volume analysis (i.e., A + A' = 40-79 microns 3; B = 80-119 microns 3; C + D greater than or equal to 120 microns 3). No differences were observed among flight (F), synchronous (SC), vivarium, and basal control groups in the A + A' (F: 28.0 +/- 3.7 vs. SC: 27.4 +/- 2.2), B (F: 33.1 +/- 1.4 vs. SC: 32.4 +/- 2.4), or C + D (F: 38.4 +/- 4.5 vs. SC: 39.2 +/- 1.6) cell compartments (mean +/- SE, n = 5). Compared with previous spaceflight experiments, the present data are consistent with a postflight response to replenish preosteoblasts and restore periodontal ligament osteogenic potential. These data emphasize the need to 1) unequivocally determine the flight effect by killing the animals in-flight and 2) further assess the postflight recovery phenomenon.

Animals↗

Physiology of osseous and fibrous integration.

The adaptation of bone to dental implants by rigid osseous integration is believed by many to be the hallmark of clinical success. However, many implants that are slightly mobile with a fibrous interface also function well in clinical application. Physiologically, why does bone respond by rigidly integrating one implant and producing a fibrous interface with another? The healing mechanism for rigid osseous integration is well known and accepted. Conversely, the development of a fibrous interface is incompletely understood. This article suggests a unified concept of the interrelationship between fibrous and osseous integration.

Alveolar Bone Loss↗