Non-decalcified Sections Research Articles

Novel Histopathological Findings of Micro Bone Fragments and Epithelial Response in the Oral Mucosa in Bisphosphonate-Related Osteonecrosis of the Jaw.

Bisphosphonate-related osteonecrosis of the jaw (BRONJ) occurs in the jawbone and interfacing oral mucosa of patients treated with bisphosphonates. Herein, we report novel histopathological findings in the oral mucosa of a surgical specimen obtained from a 61-year-old man with BRONJ. The resected jawbone and adjacent oral mucosa were separated for histological examination. The mucosal tissue was examined using Von Kossa staining and immunohistochemical (CK5/6, p63) staining of non-decalcified paraffin sections. Pseudoepitheliomatous hyperplasia (PEH), a microscopic feature of the mucosal epithelium in BRONJ, was observed in soft tissue specimens, concomitant with inflammatory cell infiltration. Von Kossa staining revealed small fragments of necrotic bone, tens to hundreds of micrometers in size, scattered within the connective tissues; the PEH forefront contacted some of the bone fragments. Immunohistochemical staining demonstrated that occasionally, the PEH not only contacted but also encompassed the bone fragments. To our knowledge, this is the first report of presence of micro bone fragments and their association with PEH in the oral mucosa in BRONJ.

Additional Insights Into the Role of Osteocalcin in Osteoblast Differentiation and in the Early Steps of Developing Alveolar Process of Rat Molars.

This study investigated whether osteocalcin (OCN) is present in osteoblast precursors and its relationship with initial phases of alveolar process formation. Samples of maxillae of 16-, 18-, and 20-day-old rat embryos (E16, E18, and E20, respectively), and 05-, 10-, and 15-day-old postnatal rats (P05, P10, and P15, respectively) were fixed and embedded in paraffin or araldite. Immunohistochemistry for osterix (Osx), alkaline phosphatase (ALP), and OCN detection was performed and the number of immunolabelled cells was computed. Non-decalcified sections were subjected to the von Kossa method combined with immunohistochemistry for Osx or OCN detection. For OCN immunolocalization, samples were fixed in 0.5% glutaraldehyde/2% formaldehyde and embedded in LR White resin. The highest number of ALP- and OCN-immunolabelled cells was observed in dental follicle of E16 specimens, mainly in basal portions of dental alveolus. In corresponding regions, osteoblasts in differentiation adjacent to von Kossa-positive bone matrix exhibited Osx and OCN immunoreactivity. Ultrastructural analysis revealed OCN immunoreactive particles inside osteoblast in differentiation, and in bone matrix associated with collagen fibrils and within matrix vesicles, at early stages of alveolar process formation. Our results indicate that OCN plays a role in osteoblast differentiation and may regulate calcium/phosphate precipitation during early mineralization of the alveolar process.

Fluoride-modified implant surfaces improves osseointegration in the tibias of rats with induced diabetes.

This study evaluated the influence of a fluoride-modified titanium surface on osseointegration in rats with induced diabetes. One hundred and eighty rats were randomly allocated into 3 groups with 60 animals each: Control group (C): Animals without diabetes; Diabetes Group (D): Animals with uncontrolled induced diabetes; Controlled Diabetes Group (CD): Animals with diabetes induced controlled by the insulin administration. Diabetes was induced by streptozotocin injection. Each animal received 2 implants in the proximal tibial metaphysis, one with the machined surface (M) and the other one with a fluoride-modified titanium surface (F), after 4 weeks of induction of diabetes. The animals were submitted to euthanasia 2, 4, and 6 weeks after the implant placement (n = 20 animals/group). The osseointegration was evaluated by the implant removal torque test and the histometric analysis of the non-decalcified histological sections: 1) Contact bone/implant (%BIC); 2) Bone tissue area between implant threads (%BBT). Implants with F surface showed a higher removal torque than implants with surface M in all groups. There was no difference in %BIC between the groups regardless of the surface used. The F surface showed a tendency to present higher %BBT values for the 3 evaluation periods in the D group. The fluoride-modified implant surface has no impact on the %BIC and %BBT. However, the fluoride-modified implant surface increases the locking of the implants with the bone. The hyperglycemia was associated with lower removal torque values despite the surfaces of the implant used.

Evaluation of hydrophilic surface osseointegration in low-density bone: Preclinical study in rabbits.

The aim of this study was to evaluate the osseointegration of a hydrophilic surface (blasting + acid etching + immersion in isotonic solution) in comparison with that of a control surface (blasting + acid etching) using an experimental model of low-density bone. To perform the study, 24 rabbits were submitted to the installation of 4 implants in the iliac bone bilaterally: 2 implants with a control surface and 2 implants with a hydrophilic surface. The rabbits were euthanized at 2, 4, and 8 weeks after implant installation. After euthanasia, one implant from each surface was used to perform the removal torque analysis, and the other implant was used for the execution of non-decalcified histological sections and evaluation of the bone implant contact (% BIC) as well as the fraction of bone tissue area between the implant threads (% BBT). The implants with a hydrophilic surface presented higher %BIC (42.92 ± 2.85% vs. 29.49 ± 10.27%) and % BBT (34.32 ± 8.52% vs. 23.20 ± 6.75%) (p < 0.05) in the 2-week period. Furthermore, the hydrophilic surface presented higher removal torque in the 8-week period (76.13 ± 16.00 Ncm2 vs. 52.77 ± 13.49 Ncm2) (p<0.05). Implants with a hydrophilic surface exhibited acceleration in the process of osseointegration, culminating in greater secondary stability in low-density bone than in implants with a control surface.

Sequential osseointegration of a novel implant system based on 3D printing in comparison with conventional titanium implants

To evaluate the sequential osseointegration of a novel titanium implant system based on a 3D printing technology in comparison with conventional titanium implants. Two novel titanium implants based on 3D printing were tested in the mandible of eight Beagle dogs. As a control, two different commercially available titanium implants were used. The implants were staged to accommodate healing periods of 2 and 6 weeks. The primary outcome variable was bone-to-implant contact (BIC) in non-decalcified tissue sections and micro-CT analysis. Histomorphometrically, the proportions of tissues adjacent to the implant surfaces were similar for all implants, whereas the BIC percentage of new mineralized bone was greater for the control implants after both 2 and 6 weeks (p < .05). Micro-CT analysis revealed increasing osseous volume and BIC from 2 to 6 weeks. In contrast to the histomorphometry, the BIC evaluation with the micro-CT data revealed a significantly higher BIC for the two test implants compared with controls (p < .001). The analysis of the total implant surface area disclosed a value that was approximately double as high for the test compared to the control implants. The novel titanium implant system based on 3D printing yielded values for osseointegration that were adequate and satisfactory. The higher percentage of new mineralized bone in the control implants is explained by the fact of a completely different three-dimensional surface area.

Bioactivated Implant Surfaces Placed in Healed Sites or Extraction Sockets: A Preliminary Experimental Study in Dogs.

To monitor the early bone reaction in a canine model to a conventional sandblasted and dual acid-etched implant surface (ABT), a nanostructured hydrophilic surface (Nano), a dry salt-bioactivated ultra-hydrophilic surface (Hydro), and a bioactivated nanosurface obtained from the addition of dry salts to the Nano surface (Nano-Active). ABT, Nano, Hydro, and Nano-Active implants were placed in 12 dogs. A randomized split-mouth design was adopted. One implant of each type was placed in the mandible 3 months after tooth extraction in healed sites at the first molar region bilaterally. In the same session, the third and fourth premolars were extracted bilaterally and one implant of each type was immediately placed into the extraction socket. The dogs were euthanized at 14 and 28 days following surgery, and the peri-implant bone reaction was assessed histologically using Stevenel's blue and alizarin red in nondecalcified sections. The postoperative healing was uneventful. The 14-day histologic analysis reported nonsignificant results in terms of difference between the groups, while significant results were found 28 days after surgery. In fact, a significantly higher rate of new bone around the implant was reported in the Nano-Active compared to the Nano groups (51.0% ± 10.2% vs 36.0% ± 10.2%) and Hydro compared to the Nano groups (47.3% ± 10.7% vs 36.0% ± 10.2%). The results obtained indicate that new bone formed after 4 weeks demonstrated a tendency for dry salt-treated bioactivated surfaces to improve bone deposition in the interface in the early stages of healing; however, due to the limited number of dogs, the results failed to show a statistical significance. A study with a significantly larger group of animals should be performed in order to challenge the assumption that ultra-hydrophilic-surface implants might show higher bone-implant contact in immediate postextraction replacement.

Morphological and morphometric changes of bone tissue in patients with osteoporosis and osteomalation

Bone tissue was studied in 56 postmenopausal women (mean age 62.30 ± 2.74 years), of which 46 patients who worked in unfavorable working conditions had a decreased bone mineral density (BMD) (osteoporosis (OP) — in 31 women, osteomalacia (OM) — in 13); 10 women had no metabolic changes in bone tissue (BT). A BT scan fragment was obtained during surgery for a fracture of the femoral neck. Non-decalcified QD sections were prepared, the functional activity of the QD cell nuclei was determined using the method of differential staining of nuclei with different functional activity. Morphological changes in OP and OM have both common features and differences. The common is the thinning of the bone rods, the expansion of the canals of osteons, the presence of cell-free areas, and cell-free lacunae. In contrast to OP, OM presents with the thickness and area of the osteoid increase, a less pronounced decrease in oxyphyllin matrix, a higher functional activity of BT cells. A decrease in BMD and the occurrence of low-energy fractures may result not only from OP but also OM. When prescribing treatment, it is necessary to carry out diffe-rential diagnostic measures that determine the cause of the decrease in bone mass.

In vivo evaluation of deer antler trabecular bone as a reconstruction material for bone defects

Availability of graft materials to fill up osseous defects has always been a concern in orthopaedic surgeries. Deer antler material is a primary bone structure that is easy to collect and could serve as a xenograft. This study examines the behaviour of red deer antler trabecular cylinders in critical size distal femoral epiphyseal defects in 11 rabbits, and evaluates the effect of the decellularization protocols.Two preparation regimes (A and B) were used, with and without lipids and proteins. Radiographs were taken immediately after surgery and after euthanasia 12 weeks post-implantation. Histological evaluation was performed on non-decalcified 10-μm sections with a van Gieson picro-fuchsin staining protocol. A region of interest was defined for each histological section, evaluating the inflammatory reaction, the fibrosis process, and the osteogenesis. Each histological section was microradiographed to evaluate bone contact, presence of synostosis, remodelling and ossification processes.All antler cylinders were successfully implanted. Final radiographic analysis demonstrated osteointegration of most implants at various stages. Light to moderate inflammation around the grafts was noted with only one case showing full encapsulation. A variable degree of intimacy between implant and host bone was evidenced, with bone remodelling and osteogenesis of various intensity being present in all implanted sites. No differences were found between group A and B.Removal of lipids and proteins in the grafts surprisingly did not seem to matter. Decellularization and sterilization protocols may be advocated. Although it presents several limitations, this study shows some promising results regarding antler trabecular bone osteointegration.

Coexistence of Osteomalacia in Osteoporotic Hip Fractures in More Than 50Years Age Group.

Osteomalacia is a hitherto common orthopaedic condition and is commonly coexists with osteoporosis. However, the identification of osteomalacia always slips under the radar and more emphasis is given to diagnosis and management of osteoporosis. Identification of osteomalacia is equally relevant as management of the osteoporotic fractures is different with or without osteomalacia. This was a prospective study design that included patients 50years or above of either sex presented with proximal femur fractures. Osteoporosis was identified by DEXA scan of hip and lumbar spine. Metabolic tests including serum calcium, phosphorus, ALP and vitamin D levels were done. Histopathological diagnosis of osteomalacia was performed on bony tissues that were taken during surgery from a site adjacent to the fracture and histological examination was performed on non-decalcified paraffin sections using special stains. A total of 45 patients was included in study. Mean age was 68.7years (53-85years). Abnormal values of serum calcium, phosphorus, ALP, vitamin D were noted in 44.4%, 22.2%, 53.3% and 48.9% patients, respectively. On histopathology, 73.17% patients showed osteomalacia. No significant correlation was found between serum biochemical markers and histopathology except with serum Vitamin D (p value - 0.004). The majority of patients with osteoporotic hip fractures had coexisting osteomalacia. Abnormal biochemical values were not significantly associated with osteomalacia. Hence, histopathology remains the gold standard for the diagnosis of osteomalacia. Further research is needed to identify a biomarker that may enable the clinician to diagnosis and treat osteomalacia well in time.

Effects of CGRP receptor antagonism on glucose and bone metabolism in mice with diet-induced obesity.

The neuropeptide calcitonin gene-related peptide (CGRP) and its receptor, calcitonin receptor-like receptor (CLR) complexing with receptor activity-modifiying protein 1 (RAMP1), have been shown to be crucially involved in the pathogenesis of migraine. However, CGRP also plays a pivotal role in regulating bone turnover and was suggested to contribute to the development of the metabolic syndrome. Therefore, our study was designed to characterize the effects of CGRP antagonism on bone and glucose metabolism in a murine model of diet-induced obesity (DIO). A subcutaneous pellet releasing the CGRP receptor antagonist BIBN 4096 (BIBN; olcegepant) was implanted in WT mice with DIO. Metabolic effects were assessed through body- and organ-weights, oral glucose tolerance (oGT), serum lipids, and gene-expression studies. Bone turnover was assessed through histomorphometry of non-decalcified bone sections and analyses of bone turnover markers in serum samples. BIBN treatment did not alter body weight gain or the levels of serum lipids including triacylglycerol and cholesterol during DIO. BIBN led to a moderate improvement of oGT which was accompanied by an increased expression of stearoyl-CoA desaturase in the liver. In skeletal tissue, BIBN treatment resulted in reduced bone volume. This was explained by decreased parameters of bone formation whereas bone resorption was not affected. Our results indicate that inhibition of CGRP signaling only moderately affects glucose metabolism during DIO but significantly impairs bone formation. As novel agents blocking CGRP or its receptor are currently introduced clinically for the treatment of migraine disorders, their potential negative impact on bone metabolism requires further clinical studies.

Osteomalacia With Hyperostosis in Captive Lesser Hedgehog Tenrecs (Echinops telfairi).

Four captive, lesser hedgehog tenrecs (Echinops telfairi) were euthanized for soft bones that prevented normal mastication and/or ambulation. Antemortem radiographs (available in 2 cases) revealed osteopenia. Antemortem bloodwork (available in 2 cases) revealed hypophosphatemia. Dietary history (available in 2 cases) indicated the animals were eating only insects. Histologically, all examined bones had wide osteoid seams that caused expansion of the cortices. Osteoid deposition was exuberant and it partially filled marrow cavities (hyperostosis). Nondecalcified sections of bone (available in 2 cases) revealed that osteoid was poorly mineralized, consistent with osteomalacia. Insects are poor dietary sources of vitamin D, and dietary vitamin D deficiency is considered the most likely cause for metabolic bone disease in these animals.

Augmentation of soft tissue volume at pontic sites: a comparison between a cross-linked and a non-cross-linked collagen matrix

AimTo assess histopathological and histomorphometric outcomes of soft tissue volume augmentation procedures at pontic sites using a volume-stable cross-linked collagen matrix (VCMX) and a non-cross-linked collagen matrix (XCM).Materials and methodsIn twelve adult beagle dogs, the mandibular premolars and first molar were hemisected and the mesial root extracted. Soft tissue augmentation was randomly performed using VCMX, XCM, or a sham-operated control. Sacrifice was performed after 4, 8, and 26 weeks. Non-decalcified sections were analyzed for histopathologic and histomorphometric measurements at four different levels below the crest (1.5, 2.5, 3.5, and 5.5 mm).ResultsGroup VCMX presented a greater overall amount of soft tissue at all healing time points, more pronounced fibroblast ingrowth, vascularization, and a substantial new collagen deposition. Over time, group XCM demonstrated faster signs of degradation compared with group VCMX. Four weeks after augmentation, group VCMX yielded a higher mean ridge width compared with groups XCM (2.22 mm VCMX, 0.89 mm XCM (at 2.5 mm); 2.05 mm VCMX, 0.80 mm XCM (at 3.5 mm) p < 0.05) and sham (0.59 mm sham (at 1.5 mm); 0.48 mm (at 2.5 mm); 0.44 mm (at 3.5 mm) p < 0.05). After healing periods of 8 and 26 weeks, measurements in group VCMX remained significantly higher compared with group sham both at 8 weeks (levels of 1.5 mm, 2.5 mm and 5.5 mm) and at 26 weeks (levels of 1.5 mm, 3.5 mm and 5.5 mm) (p < 0.05).ConclusionThe use of a cross-linked collagen matrix resulted in a greater and more stable ridge width over time compared with control groups.Clinical relevanceSoft tissue volume augmentation at pontic sites is more effective when using a cross-linked compared with a non-cross-linked collagen matrix.

Mechanism and patterns of bone loss in patients with anterior shoulder dislocation

Bony defects are common injuries associated with anterior shoulder dislocation. It is generally thought that these bony defects are created at the time of dislocation. However, there have been no biomechanical reports demonstrating the exact time point when these lesions occur. The purpose of this study was to clarify when, how, and which types of bony defects were created during experimental dislocation in cadaveric shoulders. Fifteen fresh-frozen cadaveric shoulders (mean age at the time of death, 79 years) were fixed in a custom testing machine. First, the glenohumeral joint was inspected by arthroscopy. Then, the arm was held at 60° of abduction and maximum external rotation and was manually extended horizontally under fluoroscopy until an anterior dislocation occurred. Next, a force of 800 N was applied to a Kirschner wire inserted inthe humeral head in the direction of the pectoralis major with use of an air cylinder. We waited until the arm came to equilibrium under this condition. Finally, the glenohumeral joint was arthroscopically examined. We further performed x-ray micro-computed tomography and histologicexamination in 1 shoulder with a bipolar lesion. After the anterior dislocation, a Bankart lesion was created in 9 of 15 shoulders and a fragment-type glenoid defect (avulsion fracture) was created in 4. A Hill-Sachs lesion, on the other hand, was not observed after the dislocation. The equilibrium arm position was 40° ± 17° in flexion, 45° ± 22° in abduction, and 27° ± 19° in external rotation. In this arm position, newly created lesions were Hill-Sachs lesions in 6 shoulders and erosion-type glenoid defects (compression fracture) in 7. Micro-computed tomography, performed in a single specimen, showed a flattened anterior glenoid rim with collapse of trabecular bone. Histologicanalysis of nondecalcified sections using hematoxylin-eosin staining indicated that the anterior rim of the glenoid was compressed and flattened. The cortex of the anterior glenoid rim could be clearly observed. The fragment-type glenoid defect (avulsion fracture) was observed at the time of dislocation, whereas the erosion-type defect (compression fracture) was observed when the arm came to equilibrium in the midrange of motion. Hill-Sachs lesions were created not at the time of dislocation but after the arm came to equilibrium.

Evaluation of the effects of topically applied simvastatin on titanium implant osseointegration

ObjectiveThe aim of this study was to evaluate the effect of topical simvastatin on the osseointegration of machined-surface titanium implants. Material & methodsSixteen female Sprague–Dawley rats were used in this study in a 4-wk experimental trial. The rats were divided into two groups: a test group (n = 8), in which local simvastatin was applied, and a control group (n = 8). In each of the 16 animals, a machined-surface titanium implant was surgically integrated into the tibial metaphysis. Prior to integration of the implants, 100 μl of ethanol solution containing 5 mg of simvastatin was applied to the bone sockets. The implants were then immediately integrated into the bone sockets. No simvastatin treatment was applied in the control group. At the end of the 4-wk period, the rats were sacrificed, and the implants and surrounding bone tissue were removed. The bone–implant contact (BIC) and bone filling (BF) ratios of nondecalcified histological sections were then determined. The BIC ratio was defined as the ratio of the implant surface directly in contact with the bone to the total implant surface length. The BF ratio was determined by measuring the bone-filled area at a distance of 0.5 mm from implant. ResultsThere were no statistically significant differences in the BIC and BF ratios of the test group and control group in terms of implant osseointegration (p > 0.05). ConclusionWithin the limitations of this study, we conclude that topically applied simvastatin to implant surfaces does not affect osseointegration.

Influence of Masticatory Functional Loss on the Remodeling of Alveolar Bone in Rats

There is plenty of literature on masticatory function and its impact on maxillofacial development. However, the influence of masticatory hypofunction on bone turnover in the alveolar bone has hardly been studied. This study aimed to clarify the influence of tooth loss and soft diet on the alveolar bone turnover during the growth period. Three-week-old Wistar rats were randomly divided into the following three groups: Hard diet group (rats raised on solid standard diet), Powder diet group (rats raised on powdered standard feed diet), and Extraction group (rats raised on powdered standard diet with maxillary molars extraction). BV, BMC, and BMD in the cancellous bone of M1 were measured using micro-CT analysis. To analyze the histological bone turnover, we prepared non-decalcified thin sections of alveolar cancellous bone when rats were 20 weeks old. On three-dimensional constructed images, the experimental groups (the Powder diet and Extraction groups) showed expansion of the medullary cavity of the interradicular septum of the first molar compared to controls (the Hard diet group). BV, BMC, and BMD were significantly lower in the experimental groups, with the difference from controls being greater in the Extraction group. On histomorphometric analysis, the bone mass parameters, bone formation parameters, and bone mineralization parameters were significantly lower in the experimental groups compared to controls. The bone resorption parameters were significantly higher in the experimental groups. From this study, we found that soft diet and tooth loss might worsen the bone microstructure, reduce osteogenesis, and promote bone resorption in alveolar bone.

Evaluation of Goose-beak Bone Particles for Dentoalveolar Reconstruction in Dogs.

Tooth extraction is a common procedure in dental clinics. Tooth extraction can destroy gingiva, alveolar bone, periodontal ligaments and cement. If dental sockets are left as extracted, it will result in loss of teeth, as well as voice and aesthetic problems. A natural hydroxyapatite (HA) bioceramic bone graft substitute developed from goose-beak bone particles (GBPs) was used for dentoalveolar reconstruction in a canine model. Four adult (18-22 months old) male beagle dogs weighing 8.2-9.6 kg were included in the study. Eight alveolar extraction sockets in the four dogs were divided randomly into two groups and a split-mouth design was established; control group, socket filled with commercial synthetic HA; tested group, socket filled with granulated GBP. Micro-CT analysis and hematoxylin and eosin and Masson's trichrome staining of non-decalcified sections were undertaken. Examination revealed that dentoalveolar reconstruction was initiated from the periphery of the host bone, and newly formed bone was well integrated with the GBP. Bone apposition was observed at the edge of the host bone-GBP interface. A natural ceramic powder obtained from GBP is suitable for use in dentoalveolar reconstruction in dogs.

Effect of Sintering on In Vivo Biological Performance of Chemically Deproteinized Bovine Hydroxyapatite.

The influence of the manufacturing process on physicochemical properties and biological performance of xenogenic biomaterials has been extensively studied, but its quantification on bone-to-material contact remains poorly investigated. The aim of this study was to investigate the effect of different heat treatments of an experimental chemically-deproteinized bovine hydroxyapatite in vivo in terms of new bone formation and osteoconductivity. Protein-free hydroxyapatite from bovine origin was produced under sub-critical conditions and then either sintered at 820 °C or 1200 °C. Structural and morphological properties were assessed by scanning electron microscopy (SEM), measurement of surface area and X-ray diffractometry (XRD). The materials were then implanted in standardized alveolar bone defects in minipigs and histomorphometric evaluations were performed using non-decalcified sections. Marked topographical differences were observed by SEM analysis. As the sintering temperature of the experimental material increased, the surface area significantly decreased while crystallite size increased. In vivo samples showed that the highly sintered BHA presented a significantly lower percentage of newly formed bone than the unheated one (p = 0.009). In addition, the percentage of bone-to-material contact (BMC) was significantly lowered in the highly sintered group when compared to the unsintered (p = 0.01) and 820 °C sintered (p = 0.02) groups. Non-sintered or sintered at 820 °C BHA seems to maintain a certain surface roughness allowing better bone regeneration and BMC. On the contrary, sintering of BHA at 1200 °C has an effect on its morphological and structural characteristics and significantly modify its biological performance (osteoconductivity) and crystallinity.

The effect of platelet-rich fibrin exudate addition to porous poly(lactic-co-glycolic acid) scaffold in bone healing: An in vivo study.

Bone grafting procedures have been widely utilized as the current state-of-the-art for bone regeneration, with autogenous bone graft being the gold-standard bone reconstructive option. However, the use of autografts may be limited by secondary donor-site comorbidities, a finite amount of donor supply, increased operating time, and healthcare cost impact. Synthetic materials, or alloplasts, such as the polymeric material, poly(lactic-co-glycolic acid) (PLGA) has previously been utilized as a transient scaffold to support healing of bone defects with the potential to locally delivery osteogenic additives. In this study a novel procedure was adopted to incorporate both the dissolved contents and mechanical components of leukocyte- and platelet-rich fibrin (L-PRF) into an PLGA scaffold through a two-step method: (a) extraction of the L-PRF membrane transudate with subsequent immersion of the PLGA scaffold in transudate followed by (b) delivering a fibrin gel as a low-viscosity component that subsequently polymerizes into a highly viscous, gel-like biological material within the pores of the PLGA scaffold. Two, ~0.40 cm3 , submandibular defects (n = 24) were created per side using rotary instrumentation under continuous irrigation in six sheep. Each site received a PLGA scaffold (Intra-Lock R&D, Boca Raton, FL), with one positive control (without L-PRF exudate addition [nL-PRF]), and one experimental (augmented with PLGA/L-PRF Blocks [L-PRF]). Animals were euthanized 6 weeks postoperatively and mandibles retrieved, en bloc, for histological analysis. Histomorphometric evaluation for bone regeneration was evaluated as bone area fraction occupancy (BAFO) within the region of interest of the cortical bone (with specific image analysis software) and data presented as mean values with the corresponding 95% confidence interval values. Qualitative evaluation of nondecalcified histologic sections revealed extensive bone formation for both groups, with substantially more bone regeneration for the L-PRF induced group relative nL-PRF group. Quantitative BAFO within the defect as function of the effect of L-PRF exudate on bone regeneration, demonstrated significantly (p = .018) higher values for the L-PRF group (38.26% ± 8.5%) relative to the nL-PRF group (~28% ± 4.0%). This in vivo study indicated that L-PRF exudate has an impact on the regeneration of bone when incorporated with the PLGA scaffold in a large translational model. Further studies are warranted in order to evaluate the L-PRF exudate added, as well as exploring the preparation methods, in order to facilitate bone regeneration.

Effect of Different Doses of Synthetic Parathyroid Hormone (1-34) on Bone around Implants: a Preclinical Rat Model.

The aim of this study was to evaluate the effect of a lower dose of parathyroid hormone- PTH (1-34) on osteogenic potential of bone healing around titanium implants inserted into the tibia of rats. A blind parallel study was conducted in 45 adult male Wistar rats. Each rat received one titanium implant (4.5 x 2.2 mm) and was randomly assigned to receive subcutaneous injections, three times/week for 30 days, of the following treatments: group 1 - 40 µg/kg of PTH (1-34) (n=15); group 2 - 2 µg/kg of PTH (1-34) (n=15) and; group 3 - only the vehicle required for hormone dissolution (n=15). Thirty days after surgery, the animals were sacrificed and specimens containing the implant and the surrounding bone were removed and processed for non-decalcified sections. The sections were evaluated according to the following histometric parameters: proportion of mineralized tissue (PMT) adjacent to the implant threads (500 µm band); bone filling within the limits of the threads (BF) and; bone-to-implant contact (BIC). For the cortical region, both hormone dosages (groups 1 and 2) promoted better results, for all parameters, when compared to control group (p<0.05). Similar results were observed for the BF parameter in the cancellous region (p=0.0394). Therefore, systemic administration of PTH (1-34) stimulates bone formation around titanium implants, even at low doses.

Evaluation of the Safety and Efficacy of Soft Tissue Augmentation With a Compressive-Resistant Collagen Matrix in a Nonhuman Primate Model

The purpose of the present study was to evaluate the safety and efficacy of the compression-resistant collagen-based cross-linked matrix for augmentation of maxillary and mandibular soft tissue defects in an animal model. Six rhesus macaque monkeys were subjected to soft tissue grafting in 4 sites intraorally; the anterior maxilla was subjected to hard and soft tissue grafting with implant placement. Each site was randomly assigned 1 of 3 treatments: a compressive-resistant collagen matrix membrane (CM), a subepithelial connective tissue autograft (SCTG), or sham treatment, in which a partial-thickness flap was elevated and then sutured closed with no further treatment (control). The following methods were used for data collection: invivo evaluation by periodontal probing, ultrasound, shear modulus elasticity, polyether impressions for volumetric analysis, and invitro analysis by histologic biopsy examinations. Invitro analysis provided by histologic measurements and evaluations was performed on nondecalcified sections. The follow-up period was 6months. The SCTG and CM showed favorable tissue integration. No adverse reaction to or deviation from the normal healing processes was detected. The CM integrated well in all sites, with a variable range of soft tissue volume increases. Volumetric discrepancies were appreciated in the histologic analyses and differences were found when the CM and SCTG were applied in the anterior maxilla in combination with hard tissue grafting and implant placement. Histologic evaluation showed favorable integration, no immunogenic response to the CM, and stable volumetric retention in autograft and CM sites during the experimental period. The compressive-resistant CM could be a safe and efficacious alternative for soft tissue augmentation by obviating a donor site and the consequent morbidity. Although a similar performance between the CM and SCTG was observed, further studies will be necessary to estimate the clinical potentiality and describe the limits of the technique.