Effect of platelet-rich plasma on bone healing of autogenous bone grafts in critical-size defects

AimThis study histologically analysed the effect of autogenous platelet-rich plasma (PRP), prepared according to a new semiautomatic system, on healing of autogenous bone (AB) grafts placed in surgically created critical-size defects (CSD) in rabbit calvaria.Material and MethodsSixty rabbits were divided into three groups: C, AB and AB/PRP. A CSD was created in the calvarium of each animal. In Group C (control), the defect was filled by blood clot only. In Group AB (autogenous bone graft), the defect was filled with particulate autogenous bone. In Group AB/PRP (autogenous bone graft with platelet-rich plasma), it was filled with particulate autogenous bone combined with PRP. All groups were divided into subgroups (n=10) and euthanized at 4 or 12 weeks post-operatively. Histometric and histologic analyses were performed. Data were statistically analysed (anova, t-test, p < 0.05).ResultsGroup C presented significantly less bone formation compared with Group AB and AB/PRP in both periods of analysis (p < 0.001). At 4 weeks, Group AB/PRP showed a statistically greater amount of bone formation than Group AB (64.44 +/- 15.0% versus 46.88 +/- 14.15%; p=0.0181). At 12 weeks, no statistically significant differences were observed between Groups AB and AB/PRP (75.0 +/- 8.11% versus 77.90 +/- 8.13%; p > 0.05). It is notable that the amount of new bone formation in Group AB/PRP at 4 weeks was similar to that of Group AB at 12 weeks (p > 0.05).ConclusionWithin its limitation, the present study has indicated that (i) AB and AB/PRP significantly improved bone formation and (ii) a beneficial effect of PRP was limited to an initial healing period of 4 weeks.

Repair process of surgical defects filled with autogenous bone grafts in tibiae of diabetic rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bone regeneration in surgically created defects filled with autogenous bone: an epifluorescence microscopy analysis in rats

Although the search for the ideal bone substitute has been the focus of a large number of studies, autogenous bone is still the gold standard for the filling of defects caused by pathologies and traumas, and mainly, for alveolar ridge reconstruction, allowing the titanium implants installation. Objectives: The aim of this study was to evaluate the dynamics of autogenous bone graft incorporation process to surgically created defects in rat calvaria, using epifluorescence microscopy. Material and methods: Five adult male rats weighing 200-300 g were used. The animals received two 5-mm-diameter bone defects bilaterally in each parietal bone with a trephine bur under general anesthesia. Two groups of defects were formed: a control group (n=5), in which the defects were filled with blood clot, and a graft group (n=5), in which the defects were filled with autogenous bone block, removed from the contralateral defect. The fluorochromes calcein and alizarin were applied at the 7th and 30th postoperative days, respectively. The animals were killed at 35 days. Results: The mineralization process was more intense in the graft group (32.09%) and occurred mainly between 7 and 30 days, the period labeled by calcein (24.66%). Conclusions: The fluorochromes showed to be appropriate to label mineralization areas. The interfacial areas between fluorochrome labels are important sources of information about the bone regeneration dynamics.

Influence of the proportion of particulate autogenous bone graft/platelet-rich plasma on bone healing in critical-size defects: An immunohistochemical analysis in rat calvaria

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Utilization of autogenous bone, bioactive glasses, and calcium phosphate cement in surgical mandibular bone defects in Cebus apella monkeys

Purpose: the purpose of the present study was to evaluate the histologic results of bone cavities that were surgically created in the mandibles of Cebus apella monkeys and filled with autogenous bone, PerioGlas, FillerBone, or Bone Source. Materials and Methods: Surgical cavities 5 mm in diameter were prepared through both mandibular cortices in the mandibular angle region. The cavities were randomly filled, and the animals were divided into groups according to the material employed: Group 1 cavities were filled with autogenous corticocancellous bone; group 2 cavities were filled with calcium phosphate cement (BoneSource); and group 3 and group 4 cavities were filled with bioactive glass (FillerBone and PerioGlas, respectively). After 180 days the animals were sacrificed, and specimens were prepared following routine laboratory procedures for hematoxylin/eosin staining and histologic evaluation. Results: the histologic analysis showed that autogenous bone allowed total repair of the bone defects; bioactive glasses (FillerBone and PerioGlas) allowed total repair of the defects with intimate contact of the remaining granules and newly formed bone; and the cavities filled with calcium phosphate cement (BoneSource) were generally filled by connective fibrous tissue, and the material was almost totally resorbed. Discussion: the autogenous bone, FillerBone, and PerioGlas provided results similar to those in the current literature, showing that autogenous bone is the best Choice for filling critical-size defects. Synthetic implanted materials demonstrated biocompatibility, but the bioglasses demonstrated osteoconductive activity that did not occur with calcium phosphate (BoneSource). Conclusion: According to the methodology used in this study, it can be concluded that the utilization of autogenous bone and bioactive glasses permitted the repair of surgically created critical-size defects by newly formed bone; the synthetic implanted materials demonstrated biocompatibility, and the bioactive glasses demonstrated osteoconductive activity. The PerioGlas was mostly resorbed and replaced by bone and the remaining granules were in close contact with bone; the FillerBone showed many granules in contact with the newly formed bone; BoneSource did not permit repair of the critical-size defects, and the defects were generally filled by connective fibrous tissue.

Histomorphometric evaluation of human sinus floor augmentation healing responses to placement of calcium phosphate or Ricinus communis polymer associated with autogenous bone

Background: Prosthetic rehabilitation of the posterior maxilla with dental implants is often difficult because of proximity to the maxillary sinus and insufficient bone height. Maxillary sinus floor augmentation procedures aim to obtain enough bone with an association between biomaterials and autogenous bone.Purpose: the purpose of this study was to evaluate histomorphometrically two grafting materials (calcium phosphate and Ricinus communis polymer) used in maxillary sinus floor augmentation associated with autogenous bone.Materials and Methods: Biopsies were taken from 10 consecutive subjects (mean age 45 years) 10 months after maxillary sinus floor augmentation. The sinus lift was performed with a mixture of autogenous bone and R. communis polymer or calcium phosphate in a 1:2 proportion. Routine histologic processing and staining with hernatoxylin and eosin were performed.Results: the histomorphometric analysis indicated satisfactory regenerative results in both groups for a mean of bone tissue in the grafted area (44.24 +/- 13.79% for the calcium phosphate group and 38.77 +/- 12.85% for the polymer group). Histologic evaluation revealed the presence of an inflammatory infiltrate of mononuclear prevalence that, on average, was nonsignificant. The histologic sections depicted mature bone with compact and cancellous areas in both groups.Conclusion: the results indicated that both graft materials associated with the autogenous bone were biocompatible, although both were still present after 10 months.

Histomorphometric analysis of the repair process of autogenous bone grafts fixed at rat calvaria with cyanoacrylate

Objective: The purpose of this study was to perform histological and histometric analyses of the repair process of autogenous bone grafts fixed at rat calvaria with ethyl-cyanoacrylate adhesive. Material and Methods: Thirty-two rats were divided into two groups (n=16), Group I - Control and Group II - Adhesive. Osteotomies were made at the right parietal bone for graft obtainment using a 4-mm-diameter trephine drill. Then, the bone segments were fixed with the adhesive in the parietal region of the opposite side to the donor site. After 10 and 30 days, 8 animals of each group were euthanized and the calvarias were laboratorially processed for obtaining hematoxylin and eosin-stained slides for histological and histometric analyses. Results: An intense inflammatory reaction was observed at the 10-day period. At 30 days, this reaction was less intense, despite the presence of adhesive at the recipient-site/graft interface. Graft incorporation to the recipient site was observed only at the control group, which maintained the highest graft size at 10 and 30 days. Conclusions: Although the fragment was stable, the presence of adhesive in Group II did not allow graft incorporation to the recipient site, determining a localized, discrete and persistent inflammatory reaction.

Fixation of autogenous bone grafts with ethyl-cyanoacrylate glue or titanium screws in the calvaria of rabbits

This study compared the fixation of autogenous onlay bone grafts with cyanoacrylate glue (Super Bonder) and with titanium screws. Twenty rabbits underwent bilateral parietal ostectomies. Bone segments were fixed anteriorly to the resulting bone defect. In group I, the grafts were fixed with 4 min long, 1.5 mm diameter screws; in group II, adhesive was used. The animals were killed after 5, 15, 30, 60 and 120 days. Histomorphometric analysis was used to quantify the maintenance of the graft area. Discrete areas of inflammatory reaction were seen in both groups after 5 days and for group II after 15 days. After 30 days, new bone formation was seen at the interface of the grafts. After 120 days, the graft was incorporated into the host bed in group I and partially incorporated in group II. There was a significant statistical difference regarding the mean graft areas between 15 and 120 days (p < 0.001) and between fixation methods (p < 0.002). Fixation with adhesive promoted a significantly greater area of bone graft than screw fixation, independent of time period. The adhesive was biocompatible, presented similar stability to the screw and maintained the bone area, although there was a delay in graft incorporation.

Repair of critical-size defects with autogenous periosteum-derived cells combined with bovine anorganic apatite/collagen: an experimental study in rat calvaria

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influence of different light sources and photo-activation methods on degree of conversion and polymerization shrinkage of a nanocomposite resin

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Measurement of shrinkage of composite resin by laser speckle contrast analysis

The aim of this study was to evaluate the shrinkage of a microhybrid dental composite resin photo-activated by one LED with different power densities by means of speckle technique. The dental composite resin Filtek (TM) Z-250 (3M/ESPE) at color A(2) was used for the samples preparation. Uncured composite was packed in a metallic mold and irradiated during 20 s from 100 to 1000 mW cm(-2). For the photo-activation of the samples, it was used a LED prototype (Light Emission Diode) with wavelength centered at 470 nm and adjustable power density until 1 W cm(-2). The speckle patterns obtained from the bottom composite surfaces were monitored using a CCD camera without lens. The speckle field is recorded in a digital picture and stored by CCD camera as the carrier of information on the displacement of the tested surface. The calculated values were obtained for each pair of adjacent patterns and the changes in speckle contrast as a function of time were obtained from six repeated measurements. The speckle contrasts obtained from the bottom surface with 100 mW cm(-1) were smaller than those than the other power densities. The higher power densities provided the higher shrinkage.

Red blood cells from the South American rattlesnake (Crotalus durissus terrificus) regulate volume incompletely following osmotic shrinkage and swelling in vitro

The ability of rattlesnake (Crotalus durissus terrificus) red blood cells to volume regulate in vitro has been investigated. Blood was drawn through a catheter inserted in the dorsal aorta and equilibrated to gas mixtures of different composition. Cells shrunken osmotically by increasing the extracellular osmolarity from approximate to 291 mosm l(-1) (n = 3) to approximate to 632 mosm l(-1) (calculated) only partially regulated their volume back towards the original volume either at pH 7.51 +/- 0.05 (mean +/- S.D., n = 5) or pH 7.20 +/- 0.06 (mean +/- S.D., n = 3), There was no improvement of the regulatory volume increase at low haemoglobin oxygen saturation. The limited volume restoration was inhibited by separate additions of amiloride (10(-4) M) or DIDS (10(-4) M) suggesting involvement of the Na+/H+ and Cl-/HCO3- exchangers. Cells that were swollen osmotically by an approximate to 30% dilution of the extracellular medium also exhibited a limited ability to recover their volume. Therefore, these cells show little ability to volume regulate when exposed to in vitro conditions that shrink or swell the cell. (C) 2000 Elsevier B.V. All rights reserved.

Minimal volume regulation after shrinkage of red blood cells from five species of reptiles

Red blood cells (RBCs) from most vertebrates restore volume upon hypertonic shrinkage and the mechanisms underlying this regulatory volume increase (RVI) have been studied extensively in these cells. Despite the phylogenetically interesting position of reptiles, very little is known about their red cell function. The present study demonstrates that oxygenated RBCs in all major groups of reptiles exhibit no or a very reduced RVI upon -25% calculated hyperosmotic shrinkage. Thus, RBCs from the snakes Crotalus durissus and Python regius, the turtle Trachemys scripta and the alligator Alligator mississippiensis showed no statistically significant RVI within 120 min after shrinkage, while the lizard Tupinambis merianae showed 22% volume recovery after 120 min. Amiloride (10(-4) M) and bumetanide (10(-5) M) had no effect on the RVI in T merianae, indicating no involvement of the Na(+)/H(+) exchanger (NHE) or the Na(+)/K(+)/2Cl(-) co-transporter (NKCC) or insentive transporters. Deoxygenation of RBCs from A. mississippiensis and T merianae did not significantly affect RVI upon shrinkage. Deoxygenation per se of red blood cells from T merianae elicited a slow volume increase, but the mechanism was not characterized. It seems, therefore, that the RVI response based on NHE activation was lost among the early sauropsids that gave rise to modern reptiles and birds, while it was retained in mammals. An RVI response has then reappeared in birds, but based on activation of the NKCC. Alternatively, the absence of the RVI response may represent the most ancient condition, and could have evolved several times within vertebrates. (C) 2008 Elsevier B.V. All rights reserved.

Autogenous demineralized dentin matrix for tissue engineering applications: Radiographic and histomorphometric studies

Purpose: This work evaluated the osteoconductive properties of autogenous demineralized dentin matrix (ADDM) on surgical bone defects in the parietal bone of rabbits, using the guided bone regeneration technique and polytetrafluoroethylene (PTFE) membrane. Materials and Methods: Surgical bone defects were created in 24 adult rabbits and repaired with either ADDM and PTFE (experimental group) or PTFE alone (control group). The ADDM had been obtained from the central incisors of the experimental rabbits. The rabbits were sacrificed after 15, 30, 60, and 90 days and the defects examined radiographically and histologically. Results: Radiographically, the defects in the experimental animals achieved radiopacity more quickly than the defects in the control group. Discussion: After 15, 30, 60, and 90 days of observation following surgery, the ADDM slices appeared to stimulate new bone formation. The dentin slices were completely incorporated into the new bone tissue and were resorbed during the bone repair. Conclusions: Bone repair was accelerated on the bone defects treated with ADDM when compared to the control group.