A biphasic scaffold design combined with cell sheet technology for simultaneous regeneration of alveolar bone/periodontal ligament complex

This study describes the design of a biphasic scaffold composed of a Fused Deposition Modeling scaffold (bone compartment) and an electrospun membrane (periodontal compartment) for periodontal regeneration. In order to achieve simultaneous alveolar bone and periodontal ligament regeneration a cell-based strategy was carried out by combining osteoblast culture in the bone compartment and placement of multiple periodontal ligament (PDL) cell sheets on the electrospun membrane. In vitro data showed that the osteoblasts formed mineralized matrix in the bone compartment after 21 days in culture and that the PDL cell sheet harvesting did not induce significant cell death. The cell-seeded biphasic scaffolds were placed onto a dentin block and implanted for 8 weeks in an athymic rat subcutaneous model. The scaffolds were analyzed by μCT, immunohistochemistry and histology. In the bone compartment, a more intense ALP staining was obtained following seeding with osteoblasts, confirming the μCT results which showed higher mineralization density for these scaffolds. A thin mineralized cementum-like tissue was deposited on the dentin surface for the scaffolds incorporating the multiple PDL cell sheets, as observed by H&E and Azan staining. These scaffolds also demonstrated better attachment onto the dentin surface compared to no attachment when no cell sheets were used. In addition, immunohistochemistry revealed the presence of CEMP1 protein at the interface with the dentine. These results demonstrated that the combination of multiple PDL cell sheets and a biphasic scaffold allows the simultaneous delivery of the cells necessary for in vivo regeneration of alveolar bone, periodontal ligament and cementum. © 2012 Elsevier Ltd.

Estudio histológico comparativo de la reparación ósea entre hueso alveolar y extra-alveolar en los cerdos sometidos a osteotomía con alta y baja velocidad, con refrigeración líquida

Taking into account the bone repair process in pigs has shown a greater similarity among the histological variables studied compared to other biological models, the present study has as its aim to evaluate the histological bone repair process of osteotomy performed on alveolar and extra-alveolar bones, using drilling tools with liquid refrigeration. Material and method: Eighteen Large White pigs weighing between 20 and 25 Kg were divided into three groups of six animals in order to evaluate the osteotomy repairs with low and high speed in the alveolar bone and threes in the extra-alveolar area, study periods of 7, 14 and 28 days. Results: It was observed that in the alveolar bone at the postoperative times of 14 and 28 days, the best repair results were in the osteotomy performed with low speed, while in the 7 day postoperative period, the results with high speed were slightly better, in alveolar areas as well as extra-alveolar areas. There no statistically significant differences between the alveolar and extra-alveolar bone repair process. Conclusions: The repair process, by means of microscopic analysis in the alveolar and extra-alveolar areas, are similar with better results observed in osteotomies performed with low speed drills in the 14 and 28 day study periods, and at 7 days postoperative the results with high speed drills and refrigeration were slightly better. Research works using pigs as an animal model are perfectly viable. © 2011 SECOM.

Stress distribution in delayed replanted teeth splinted with different orthodontic wires: a three-dimensional finite element analysis

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

Estrogen deficiency-associated bone loss in the maxilla: A methodology to quantify the changes in the maxillary intra-radicular alveolar bone in an ovariectomized rat osteoporosis model

The effects of estrogen deficiency on bone characteristics are site-dependent, with the most commonly studied sites being appendicular long bones (proximal femur and tibia) and axial bones (vertebra). The effect on the maxillary and mandibular bones is still inconsistent and requires further investigation. This study was designed to evaluate bone quality in the posterior maxilla of ovariectomized rats in order to validate this site as an appropriate model to study the effect of osteoporotic changes. Methods: Forty-eight 3-month-old female Sprague-Dawley rats were randomly divided into two groups: an ovariectomized group (OVX, n=24) and Sham-operated group (SHAM, n=24). Six rats were randomly sacrificed from both groups at time points 8, 12, 16 and 20 weeks. The samples from tibia and maxilla were collected for Micro CT and histological analysis. For the maxilla, the volume of interest (VOI) area focused on the furcation areas of the first and second molar. Trabecular bone volume fraction (BV/TV, %), trabecular thickness (Tb.Th.), trabecular number (Tb.N.), trabecular separation (Tb.Sp.), and connectivity density (Conn.Dens) were analysed after Micro CT scanning. Results: At 8 weeks the indices BV/TV, Tb.Sp, Tb.N and Conn.Dens showed significant differences (P<0.05) between the OVX and SHAM groups in the tibia. Compared with the tibia, the maxilla developed osteoporosis at a later stage, with significant changes in maxillary bone density only occurring after 12 weeks. Compared with the SHAM group, both the first and second molars of the OVX group showed significantly decreased BV/TV values from 12 weeks, and these changes were sustained through 16 and 20 weeks. For Tb.Sp, there were significant increases in bone values for the OVX group compared with the SHAM group at 12, 16 and 20 weeks. Histological changes were highly consistent with Micro CT results. Conclusion: This study established a method to quantify the changes of intra-radicular alveolar bone in the posterior maxilla in an accepted rat osteoporosis model. The degree of the osteoporotic changes to trabecular bone architecture is site-dependent and at least 3 months are required for the osteoporotic effects to be apparent in the posterior maxilla following rat OVX.

Influence of chronic alcoholism and oestrogen deficiency on the variation of stoichiometry of hydroxyapatite within alveolar bone crest of rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of different durations of estrogen deficiency on alveolar bone loss in rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of Long-Term FK 506 Therapy on the Alveolar Bone and Cementum of Rats

Cyclosporine (CsA) and tacrolimus (FK 506) exert complex, incompletely understood actions on bone. The objective of the study was to evaluate the effects of long-term tacrolimus therapy on the periodontium. Rats were treated for 60, 120, 180, and 240 days with daily subcutaneous injections of 1 mg/kg body weight of FK 506. After the experimental period, we obtained serum levels of calcium and alkaline phosphatase (ALP). After histological processing, the alveolar bone and cementum, as well as volume densities of bone (Vb) and osteoclasts (Vo), were assessed at the regions of the lower first molar. There was a tendency toward a statistically significant decrease in ALP levels with FK 506; however, serum calcium levels increased during the long periods. At 60, 180, and 240 days of treatment with FK 506, we did not observe Vb and Vo alterations. At 120 days of treatment, there was an evident decrease in Vb, but it did not show alveolar bone loss. We did not observe any alterations of cementum among rats treated with FK 506. It may be concluded that FK 506 administration did not induce side effects on the periodontium. © 2009 Elsevier Inc. All rights reserved.

Low caloric value of ethanol itself increases alveolar bone loss in ligature-induced periodontitis in male rats

This study aimed at morphometrically evaluating the influence of variable caloric values of ethanol consumption on alveolar bone loss in periodontitis in male rats. Thirty-six male rats were randomized into four groups of nine rats each, as follows: Test group A (low) - rats were fed an ethanol-containing liquid diet (ethanol representing 22% of total caloric value); Control group A - rats were fed a pair-fed control diet (ethanol replaced by isocaloric amounts of carbohydrate); Test group B (high) - rats were fed an ethanol-containing liquid diet (ethanol representing 36% of total caloric value); Control group B - rats were fed a pair-fed control diet for Test B. Following anesthesia, cotton ligatures were placed around the cervix of the right upper second molar. At eight weeks, the maxillary bones were removed and alveolar bone loss was analyzed by measuring the distance between the cementoenamel junction and the alveolar bone crest at buccal and palatal sites of the upper second molar. The unligated groups showed no significant differences between the bone loss values observed for the low and high caloric values of ethanol (p > 0.05). In the ligated groups, the rats receiving low caloric values of ethanol showed significantly greater bone loss compared to the isocaloric rats (p < 0.05); however, the rats receiving high caloric values of ethanol showed no significant differences compared to the controls. Analysis of the results demonstrated that, in male rats, ethanol itself affected ligature-induced bone loss when representing a low value in the total caloric value.

Influence of alcohol consumption on alveolar bone level associated with ligature-induced periodontitis in rats

Alcohol consumption is a risk indicator for periodontal disease. The purpose of this study was to morphometrically evaluate the influence of alcohol consumption on alveolar bone level associated with ligature-induced periodontitis in rats. Thirty-six female rats (Wistar, 120 days-old) were randomly divided into three groups that received a daily administration of a water diet (control, n = 12), a 10% alcohol diet (10% ethanol, n = 12) or a 20% alcohol diet (20% ethanol, n = 12). Four weeks after the onset of the experiment, cotton ligatures were placed around the cervix of the upper right second molar in six rats. The other 6 rats in each group remained unligated. The rats were sacrificed four weeks after ligature placement. The maxillary bones were removed and alveolar bone loss was analyzed by measuring the distance between the cementoenamel junction and the alveolar bone crest at 2 buccal and 2 palatal sites on the upper right second molar. Analyses between the ligated and unligated groups showed that the presence of ligature induced alveolar bone loss (p < 0.05). Unligated groups showed no significant differences between each other (p > 0.05). In the ligated groups, rats receiving 20% ethanol showed significantly greater bone loss compared to control rats or rats receiving 10% ethanol. These results demonstrate that alcohol consumption may increase alveolar bone loss in female rats in a dosedependent manner.

Influência do alcoolismo crônico e da deficiência estrogênica sobre a variação de estequiometria da hidroxiapatita na crista óssea alveolar de ratas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Alcoholic and isocaloric diet, but not ovariectomy, influence the apoptosis of bone cells within the alveolar bone crest of rats

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

Alveolar ridge augmentation with the perforated and nonperforated bone grafts

Purpose: Autogenous bones are frequently used because of their lack of antigenicity, but good osteoconductive and osteoinductive properties. This study evaluated the biological behavior of perforated and nonperforated cortical block bone grafts.Methods: Ten nonsmoking patients who required treatment due to severe resorption of the alveolar process and subsequent implant installation were included in the study. The inclusion criteria was loss of one or more teeth; the presence of atrophy of the alveolar process with the indication of reconstruction procedures to allow rehabilitation with dental implants; and the absence of systemic disease, local infection, or inflammation. The patients were randomly divided into two groups based on whether they received a perforated (inner surface) or nonperforated graft. After a 6-month healing period, a biopsy was performed and osseointegrated implants were installed in the same procedure.Results: Fibrous connective tissue was evident at the interface in patients who received nonperforated grafts. However, full union between the graft and host bed was visible in those who had received a perforated graft.Conclusions: We found that cortical inner side perforations at donor sites increased the surface area and opened the medullary cavity. Our results indicate an increased rate of graft incorporation in patients who received such perforated grafts.

Fatal alveolar echinococcosis of the lumbar spine

For the last 10 years, the southern part of Belgium has been recognized as a low-risk area of endemicity for alveolar echinococcosis. This infection, caused by Echinococcus multilocularis, usually induces a severe liver condition and can sometimes spread to other organs. However, alveolar echinococcosis involving bones has been described only very rarely. Here, a fatal case of spondylodiscitis due to E. multilocularis contracted in southern Belgium is reported.

Comparison of human alveolar osteoblasts cultured on polymer-ceramic composite scaffolds and tissue culture plates

The effects of medical grade polycaprolactone–tricalcium phosphate (mPCL–TCP) (80:20) scaffolds on primary human alveolar osteoblasts (AOs) were compared with standard tissue-culture plates. Of the seeded AOs, 70% adhered to and proliferated on the scaffold surface and within open and interconnected pores; they formed multi-layered sheets and collagen fibers with uniform distribution within 28 days. Elevation of alkaline phosphatase activity occurred in scaffold–cell constructs independent of osteogenic induction. AO proliferation rate increased and significant decrease in calcium concentration of the medium for both scaffolds and plates under induction conditions were seen. mPCL–TCP scaffolds significantly influenced the AO expression pattern of osterix and osteocalcin (OCN). Osteogenic induction down-regulated OCN at both RNA and protein level on scaffolds (3D) by day 7, and up-regulated OCN in cell-culture plates (2D) by day 14, but OCN levels on scaffolds were higher than on cell-culture plates. Immunocytochemical signals for type I collagen, osteopontin and osteocalcin were detected at the outer parts of scaffold–cell constructs. More mineral nodules were found in induced than in non-induced constructs. Only induced 2D cultures showed nodule formation. mPCL–TCP scaffolds appear to stimulate osteogenesis in vitro by activating a cellular response in AO's to form mineralized tissue. There is a fundamental difference between culturing AOs on 2D and 3D environments that should be considered when studying osteogenesis in vitro.