The impact of burial period on compact bone microstructure: Histological analysis of matrix loss and cell integrity in human bones exhumed from tropical soil.

Human bone histological analysis is a useful tool to assess post mortem diagenesis and to predict successful nuclear DNA typing of forensic material. This study is part of a series of studies developed by the authors intended to improve the understanding of post mortem diagenesis and to develop applications for DNA analysis of skeletal species from tropical soils, in order to optimize genetic and anthropological protocols. The aim of this study was to analyze the impact of burial period on the integrity of exhumed compact bone microstructure from tropical climate. In fragments of exhumed human femora from 39 individuals from the same cemetery (exhumed group) and 5 fresh femora from routine autopsies (control group), sections stained by hematoxylin-eosin were analyzed in order to measure bone microstructural integrity. We found that bone integrity index in exhumed group was negatively influenced by the period of burial (r = -0.37, p < 0.05) and highly significantly decreased (p < 0.0001) in comparison to control group. The period of burial and nitric acid decalcification time was positively correlated (r = 0.51; p < 0.01), leading to imply a bone petrification process during inhumation. Exhumed group showed higher level of matrix bone loss (p < 0.001), as expected, and 87% of cases analyzed were "tunneled" as described by Hackett. Bone integrity index and bone matrix tend to decrease in bones buried in tropical soil between 8-14 years of inhumation. This period is short if we consider cases in which there are preserved bones interred for longer periods in other environments. These data must be considered in cases where genetic identification of exhumed skeletons from tropical environment is required. The diagenesis in these bones and the variations of results found are discussed, clarifying some challenges for forensic laboratories, especially in DNA analysis.

Leukocyte-platelet-rich plasma diminishes bone matrix deposition in rat calvaria treated with autograft due to simultaneous increase in immunohistochemical expression of Indian Hedgehog, transforming growth factor-ß, and parathyroid-1 receptor.

PURPOSE: This study evaluated the immunohistochemical presence of Indian Hedgehog (IHH), transforming growth factor-ß (TGF-ß), and parathyroid-1 receptor (PTH1R) in calvaria bone repair, and compared these results with the histological bone matrix features in defects treated with autograft in the presence or absence of L-PRP. MATERIAL AND METHODS: An artificial bone defect measuring 5 × 1 mm was produced in the calvaria of 28 Wistar rats. Randomly the defects were treated with autograft and autograft mixed with L-PRP. The animals were euthanized at 15 and 40 days post-surgery. Data were analyzed by Student-Newman-Keuls test (p ≤ .05) for immunohistochemical interpretation. RESULTS: The results revealed that the histological characteristic of bone matrix deposited in the defect was different in the defects treated with L-PRP. The group that received only the autograft demonstrated larger haversian bone matrix deposited, whereas the group that received autograft mixed with L-PRP revealed trabecular bone deposition. These results coincided with significantly higher immunopositivity for IHH, TGF-ß1, and PTH1R in the L-PRP group. CONCLUSION: These results suggest that L-PRP altered the biological characteristic of the autograft, increasing the bone cells IHH+ but inducing a trabecular bone associated with intense quantities of TGF-ß and PTH1R.

Use of resorbable screws for autogenous onlay block graft fixation: a histological analysis in rabbits.

The aim of the present in vivo study is to histologically evaluate and compare the use of resorbable screws based on poly(L-co-D,L lactide) 70:30 for fixation of autogenous bone grafts in rabbit tibiae. As control group, titanium (Ti-6Al-4V Grade V) screws were used. For this purpose, 15 white New Zealand male rabbits, aged 6 months and weighing between 3.8 and 4.5 kg, were used. From each animal, 2 total-thickness bone grafts were removed from the cranial vault: one was stabilized with a resorbable screw while the other was stabilized with a metallic one. Animals were divided into 3 groups, according to the sacrifice period: 3, 8, and 16 weeks postoperatively. After histological processing, cuts were stained with hematoxylin and eosin and submitted for descriptive histological analysis under light microscopy. It was found that the fixation system based on the polymer showed a histological behavior similar to metallic screws. For both groups, the bone graft was incorporated, with the presence of bone formation between the graft and receptor site. In none of the groups were undesirable inflammatory responses or foreign body reactions observed. Based on histological findings and on this experimental model, it is possible to conclude that the internal fixation system based on the poly(L-co-D,L lactide) 70:30 polymer is effective for fixation of autogenous bone grafts, with results that are comparable to the titanium fixation system.

Comparison of biomaterial implants in the dental socket: histological analysis in dogs.

BACKGROUND: Bone graft procedures have been used commonly in buco-maxillo-facial surgery. For this reason, many researchers have evaluated the bone substitutes. PURPOSE: The present study evaluated soft and hard tissue reactions to two different hydroxyapatites HAs (synthetic HA and natural HA) and bioactive glass implanted into the sockets immediately after extraction. MATERIALS AND METHODS: First and third upper and lower premolars, on both sides, were extracted from six female dogs. The alveolar sockets were randomly assigned to four groups: Group 1 - control (unfilled), Group 2 - filled with synthetic hydroxyapatite, Group 3 - filled with bovine bone mineral (natural HA), and Group 4 - filled with bioactive glass. The animals were euthanized at 4 weeks (n = 2), 8 weeks (n = 2), and 28 weeks (n = 2) after extraction. The mandible and maxilla of each animal were removed for histological analysis to determine soft tissue reactions, newly formed bone, bone characteristics, and presence or absence of implanted materials. RESULTS: Most particles of synthetic hydroxyapatite had bone formation on their surface, although some particles showed a layer of fibrous connective tissue. The bovine bone mineral group exhibited particles partially replaced with bone formation. The bioactive glass group showed particles with a thin layer of calcified tissue, but was absent in some specimens, suggesting complete resorption. CONCLUSION: All biomaterials had similar behavior. Bovine bone mineral, compared to synthetic hydroxyapatite and bioactive glass, showed a larger number of particles covered with osseous tissue. All biomaterials interfered with the socket repair process.

Histological and histomorphometric analysis of the bone-screw interface in the mandibular body after using a 2.0-mm miniplate system: an experimental study in dogs.

PURPOSE: To evaluate the bone-screw interface in a 2.0-mm miniplate system used for rigid internal fixation (RIF). MATERIALS AND METHODS: Nine adult mongrel dogs were subjected to unilateral continuous defect through an osteotomy between the lower third and fourth premolars. The control contralateral side remained untreated. Two 4-hole miniplates were placed bilaterally according to the Arbeitgeimeinschaft für Osteosynthesefragen manual. Miniplates adapted to the alveolar processes were fixed monocortically with 6.0-mm-long titanium alloy screws, whereas miniplates placed near the mandible base were fixed bicortically. At 2, 6, and 12 weeks, mandible segments enclosing the RIF were removed, fixed in formalin, ground-sectioned, and stained with toluidine blue. Under conventional light microscopy, proportions of bone-to-screw contact (BSC) were determined, and data were compared by analysis of variance. RESULTS: At 2 weeks, for both groups, the area between threads exhibited necrotic bone with multiple cracks and absence of bone cells and blood vessels. At 6 and 12 weeks, new Haversian systems progressively replaced necrotic bone. At each time point, no significant differences were seen between screws placed in the same miniplate or between groups. The proportions of BSC were statistically similar at 2 and 6 weeks and lower at 12 weeks. CONCLUSION: The results suggested that in this model, osteotomy did not significantly alter bone repair adjacent to the screw surface.

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 hematoxylin 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 homogenous demineralized dentin matrix as osteopromotive material in rabbit mandibles.

PURPOSE: The aim of this work was to evaluate the effectiveness of homogenous demineralized dentin matrix (HDDM) slices in surgical bone defects created in the mandibles of rabbits and occluded with a polytetrafluoroethylene (PTFE) membrane in the promotion of bone growth. MATERIALS AND METHODS: Surgical bone defects were created in 36 adult rabbits and divided into 4 groups: bone defect (control), bone defect with PTFE membrane, bone defect with HDDM, and bone defect with both HDDM and a PTFE membrane (HDDM + PTFE). The rabbits were sacrificed after 30, 60, and 90 days, and the bone defects were examined histologically and by histomorphometric analysis (analysis of variance and the Tukey test). RESULTS: The volume of newly formed bone matrix was significantly greater in the HDDM and HDDM + PTFE groups than in the control and PTFE groups. The discrete inflammatory reaction found in the HDDM and HDDM + PTFE groups did not prevent the osteopromotive activity of the dentin matrix. DISCUSSION: HDDM slices were biocompatible and were resorbed during the bone remodeling process. They stimulated the newly formed bone until 30 days after implantation. CONCLUSION: Bone repair was accelerated in the bone defects treated with HDDM in comparison to the control group.