Smart Delivery of Biomolecules Interfering with Peri-Implant Repair in Osteoporotic Rats.

Bisphosphonates are widely used for the treatment of postmenopausal osteoporosis; however, they cause several long-term side effects, necessitating the investigation of local ways to improve osseointegration in compromised bone tissue. The purpose of this study was to evaluate peri-implant bone repair using implants functionalized with zoledronic acid alone (OVX ZOL group, n = 11), zoledronic acid + teriparatide (OVX ZOL + TERI group, n = 11), and zoledronic acid + ruterpy (OVX ZOL + TERPY group, n = 11) compared to the control group (OVX CONV, n = 11). Analyses included computer-assisted microtomography, qualitative histologic analysis, and real-time PCR analysis. Histologically, all functionalized surfaces improved peri-implant repair, with the OVX ZOL + TERI group standing out. Similar results were found in computerized microtomography analysis. In real-time PCR analysis, however, the OVX ZOL and OVX ZOL + TERPY groups showed better results for bone formation, with the OVX ZOL + TERPY group standing out, while there were no statistical differences between the OVX CONV and OVX ZOL + TERI groups for the genes studied at 28 postoperative days. Nevertheless, all functionalized groups showed a reduced rate of bone resorption. In short, all surface functionalization groups outperformed the control group, with overall better results for the OVX ZOL + TERI group.

Analysis of peri-implant bone tissue between hydrophilic and rough implant surfaces in spontaneously hypertensive rats treated with losartan.

OBJECTIVES: to evaluate the morphological and functional characteristics of the peri-implant bone tissue that was formed during the healing process by the placement implants using two different surface treatments: hydrophilic Acqua™ (ACQ) and rough NeoPoros™ (NEO), in spontaneously hypertensive (SHR) and normotensive rats (Wistar) whether or not treated with losartan. METHODOLOGY: In total, 96 male rats (48 Wistar and 48 SHR) were divided into eight subgroups: absolute control rough (COA NEO), absolute control hydrophilic (COA ACQ), losartan control rough (COL NEO), losartan control hydrophilic (COL ACQ), SHR absolute rough (SHR NEO), SHR absolute hydrophilic (SHR ACQ), SHR losartan rough (SHRL NEO), and SHR losartan hydrophilic (SHRL ACQ). The rats medicated with losartan received daily doses of the medication. NeoPoros™ and Acqua™ implants were installed in the tibiae of the rats. After 14 and 42 days of the surgery, the fluorochromes calcein and alizarin were injected in the rats. The animals were euthanized 67 days after treatment. The collected samples were analyzed by immunohistochemistry, biomechanics, microcomputerized tomography, and laser confocal scanning microscopy analysis. RESULTS: The osteocalcin (OC) and vascular endothelium growth factor (VEGF) proteins had moderate expression in the SHRL ACQ subgroup. The same subgroup also had the highest implant removal torque. Regarding microarchitectural characteristics, a greater number of trabeculae was noted in the control animals that were treated with losartan. In the bone mineralization activity, it was observed that the Acqua™ surface triggered higher values of MAR (mineral apposition rate) in the COA, COL, and SHRL groups (p<0.05). CONCLUSION: the two implant surface types showed similar responses regarding the characteristics of the peri-implant bone tissue, even though the ACQ surface seems to improve the early stages of osseointegration.

Morphological and biomechanical characterization of long bones and peri-implant bone repair in type 2 diabetic rats treated with resveratrol.

Type 2 diabetes interferes with bone remodeling mechanisms, requiring studies to reverse this damage, and resveratrol is a polyphenol with rich properties. This study aimed to characterize the long bone morphology and peri-implant biomechanics of normoglycemic and type 2 diabetic animals treated with resveratrol. Thirty-two male Wistar rats were used and divided into normoglycemic and diabetic with or without treatment. They had the installation of implants in the tibia and treatment with oral resveratrol within 45 days. Resveratrol was responsible for weight homeostasis and decreased glycemic levels in rats with type 2 diabetes. The three-point bending testing, resveratrol showed positive effects on the biomechanics of long bones, corroborating a more resistant bone in comparison to untreated diabetics. Micro-ct revealed how bone metabolism is affected by systemic disease, decreasing bone quality. The counter-torque of normoglycemic animals showed superior osseointegration to diabetes, with no differences in the administration of the polyphenol, showing the sovereignty of the deleterious effects of the disease when there is a tissue lesion and an inflammatory picture installed. Overall, resveratrol acted positively in the etiopathogenesis of type 2 diabetes and revealed positive effects on the strength of long bones.

Analysis of peri-implant bone tissue between hydrophilic and rough implant surfaces in spontaneously hypertensive rats treated with losartan

Abstract Hypertensive individuals present alterations in their calcium metabolism that consequently decrease the concentration of this mineral in the bone tissue. Objectives to evaluate the morphological and functional characteristics of the peri-implant bone tissue that was formed during the healing process by the placement implants using two different surface treatments: hydrophilic Acqua™ (ACQ) and rough NeoPoros™ (NEO), in spontaneously hypertensive (SHR) and normotensive rats (Wistar) whether or not treated with losartan. Methodology In total, 96 male rats (48 Wistar and 48 SHR) were divided into eight subgroups: absolute control rough (COA NEO), absolute control hydrophilic (COA ACQ), losartan control rough (COL NEO), losartan control hydrophilic (COL ACQ), SHR absolute rough (SHR NEO), SHR absolute hydrophilic (SHR ACQ), SHR losartan rough (SHRL NEO), and SHR losartan hydrophilic (SHRL ACQ). The rats medicated with losartan received daily doses of the medication. NeoPoros™ and Acqua™ implants were installed in the tibiae of the rats. After 14 and 42 days of the surgery, the fluorochromes calcein and alizarin were injected in the rats. The animals were euthanized 67 days after treatment. The collected samples were analyzed by immunohistochemistry, biomechanics, microcomputerized tomography, and laser confocal scanning microscopy analysis. Results The osteocalcin (OC) and vascular endothelium growth factor (VEGF) proteins had moderate expression in the SHRL ACQ subgroup. The same subgroup also had the highest implant removal torque. Regarding microarchitectural characteristics, a greater number of trabeculae was noted in the control animals that were treated with losartan. In the bone mineralization activity, it was observed that the Acqua™ surface triggered higher values of MAR (mineral apposition rate) in the COA, COL, and SHRL groups (p<0.05). Conclusion the two implant surface types showed similar responses regarding the characteristics of the peri-implant bone tissue, even though the ACQ surface seems to improve the early stages of osseointegration.

Synergic Action of Systemic Risedronate and Local Rutherpy in Peri-implantar Repair of Ovariectomized Rats: Biomechanical and Molecular Analysis.

Postmenopausal osteoporosis and poor dietary habits can lead to overweightness and obesity. Bisphosphonates are the first-line treatment for osteoporosis. However, some studies show that they may increase the risk of osteonecrosis of the jaw. Considering the antimicrobial, angiogenic and vasodilatory potential of nitric oxide, this study aims to evaluate the local activity of this substance during the placement of surface-treated implants. Seventy-two Wistar rats were divided into three groups: SHAM (SHAM surgery), OVX + HD (ovariectomy + cafeteria diet), and OVX + HD + RIS (ovariectomy + cafeteria diet + sodium risedronate treatment), which were further subdivided according to the surface treatment of the future implant: CONV (conventional), TE10, or TE100 (TERPY at 10 or 100 µM concentration); n = 8 per subgroup. The animals underwent surgery for implant installation in the proximal tibia metaphysis and were euthanized after 28 days. Data obtained from removal torque and RT-PCR (OPG, RANKL, ALP, IBSP and VEGF expression) were subjected to statistical analysis at 5% significance level. For biomechanical analysis, TE10 produced better results in the OVX + HD group (7.4 N/cm, SD = 0.6819). Molecular analysis showed: (1) significant increase in OPG gene expression in OVX groups with TE10; (2) decreased RANKL expression in OVX + HD + RIS compared to OVX + HD; (3) significantly increased expressions of IBSP and VEGF for OVX + HD + RIS TE10. At its lowest concentration, TERPY has the potential to improve peri-implant conditions.

Estrogen Deficiency Impairs Osseointegration in Hypertensive Rats Even Treated with Alendronate Coated on the Implant Surface.

Hypertension and estrogen deficiency can affect bone metabolism and therefore increase the risk of osseointegration. Antihypertensive drugs such as losartan not only control blood pressure but also enhance bone healing. In addition, alendronate sodium is widely used to treat postmenopausal osteoporosis. Hence, we evaluated the effect of systemic antihypertensive and local alendronate coted on implants on osseointegration under hypertensive and estrogen-deficiency conditions. A total of 64 spontaneously hypertensive rats (SHRs) treated with losartan were randomly divided according to the estrogen-deficiency induction by ovariectomy (OVX) or not (SHAM), and whether the implant surface was coated with sodium alendronate (ALE) or not, resulting in four groups: SHR SHAM, SHR SHAM ALE, SHR OVX, and SHR OVX ALE. The removal torque, microcomputed tomography, and epifluorescence microscopy were the adopted analyses. The hypertensive and estrogen-deficiency animals presented a lower removal torque even when treated with alendronate on implant surface. The microcomputed tomography revealed a higher bone volume and bone-to-implant contact in the SHRs than the SHR OVX rats. Epifluorescence showed a decreased mineral apposition ratio in the SHR OVX ALE group. The data presented indicate that estrogen deficiency impairs osseointegration in hypertensive rats; in addition, alendronate coated on the implant surface does not fully reverse this impaired condition caused by estrogen deficiency.

PTH 1-34-functionalized bioactive glass improves peri-implant bone repair in orchiectomized rats: Microscale and ultrastructural evaluation.

The objective of this work was to investigate the use of Biogran® functionalized with parathyroid hormone (PTH) 1-34 by sonochemistry for the local delivery of this anabolic agent to the implant site. The effects of Biogran® and topical administration of PTH 1-34 on peri-implant bone regeneration were evaluated from the microscale to ultrastructural levels in healthy (SHAM) and orchiectomized (ORQ). While some animals only received a titanium implant in their tibial metaphyses (CLOT group), in others the peri-implant defect was first filled with Biogran® either without or with PTH 1-34 functionalization (BG and BGPTH groups, respectively) prior to implant installation. Osseointegration was characterized from a biomechanical perspective by measuring the removal torque with the counter-torque technique. Micro-CT was used to evaluate the percentage of bone volume, trabecular thickness, number and separation, and bone-implant contact (BIC). Dynamics of new bone formation were assessed by measuring fluorochrome area, daily mineral apposition rate, and neoformed bone area using confocal laser microscopy. RT-PCR was performed to evaluate ALP and osteocalcin expression. The interface between newly formed bone and Biogran® was examined using scanning electron microscopy (SEM) and scanning transmission electron microscopy (STEM) at the micro-and nanoscale, respectively, while elemental analyses were completed in SEM with energy-dispersive X-ray spectroscopy (EDS). STEM imaging demonstrated the intimate attachment of bone to Biogran® (nanoscale level). Overall, the results suggest that the effectiveness of the topical administration of PTH 1-34 at the implant site seems enhanced in osteoporotic bone, promoting peri-implant bone regeneration to comparable levels in healthy conditions.

Improvement of Peri-Implant Repair in Estrogen-Deficient Rats Fed a Cafeteria Diet and Treated with Risedronate Sodium.

(1) Background: Postmenopausal osteoporosis combined with an unhealthy lifestyle can lead to the development of metabolic syndrome, a common condition in individuals requiring oral rehabilitation. Bisphosphonates are used to increase bone mineral density. However, further studies are needed to evaluate the action of this drug on the bone repair process in the jaws. The aim of this study was to evaluate the peri-implant repair of rats with estrogen deficiency and metabolic syndrome treated with risedronate sodium. (2) Methods: Twenty-four female Wistar rats were divided into three groups: SHAM: sham surgery; OVX/SM: ovariectomy combined with a cafeteria diet; OVX/SM/RIS: ovariectomy associated with a cafeteria diet and treatment with sodium risedronate. After 30 days, the animals underwent extraction of the upper first molars. Thirty days after the extraction, an implant was installed in the same region. Sixty days after the implant was installed, the animals were euthanized for biomechanical analysis and confocal microscopic analysis. After confirming the normal distribution of the sample data, a one-way ANOVA test was performed, followed by Tukey's post-test, with a 5% significance level. (3) Results: Significant bone preservation was observed in the risedronate-treated group. Higher removal torque values were obtained by the risedronate-treated group. (4) Conclusions: Better biomechanical performance of the implants installed in the animals treated with risedronate sodium was observed.

Effect of Topical PTH 1-34 Functionalized to Biogran® in the Process of Alveolar Repair in Rats Submitted to Orchiectomy.

(1) Background: There are many therapies for osteoporosis control and bone maintenance; anabolic drugs such as teriparatide and bone grafts help in the repair process and stimulate bone formation. Thus, the aim of the present study was to evaluate the behavior of repaired bone in the presence of PTH (teriparatide) associated with Biogran® (biomaterial) through a sonochemical procedure after extraction in rats. (2) Methods: The insertion of Biogran® with PTH in the alveolus was performed 30 days after incisor extraction. Euthanasia occurred after 60 days. (3) Results: The use of local treatment of PTH loaded with Biogran® in healthy rats promoted good results for micro-CT, with an increase in percentage and bone volume, number and trabecular separation and less total porosity. Greater immunostaining for Wnt, ß-Catenin and osteocalcin proteins and lower expression for Thrombospondin-Related Adhesive Protein (TRAP), which shows an increase in the number of osteoblasts and inhibition of osteoclast action. However, the treated orchiectomized groups did not obtain such expressive results. (4) Conclusion: The use of Biogran® with PTH improved alveolar repair in rats. However, new researches with more efficient doses must be studied to collaborate effectively with the formation of a quality bone after the orchiectomy.

Caracterização do tecido ósseo peri-implantar frente ao uso do óxido nítrico para funcionalização da superfície de implantes instalados na maxila de ratas induzidas à deficiência de estrógeno e síndrome metabólica tratadas com risedronato de sódio / Characterization of peri-implant bone tissue against the use of nitric oxide for surface functionalization of implants installed in the maxilla of rats induced to estrogen deficiency and metabolic syndrome treated with risedronate sodium

Objetivo: caracterizar o processo de reparo ósseo peri-implantar em maxilas de ratas ovariectomizadas com síndrome metabólica induzida por dieta de cafeteria, tratadas com risedronato e instalação de implantes funcionalizados com [Ru(terpy)(bdq)NO]3+ (TERPY). Materiais e métodos: os testes in vitro realizados na Fase 1 avaliou as propriedades biológicas e físico-químicas da melhor concentração da TERPY frente às respostas osteogênicas, para a funcionalização dos implantes pela técnica de LbL. A realização de experimentos in vivo na Fase 2 avaliou o efeito da superfície funcionalizada durante o reparo ósseo peri-implantar. Para isso, 48 ratas Wistar foram divididas em: SHAM CONV (n=8), OVX SM CONV (n=8), OVX SM RIS CONV (n=8), SHAM TERPY (n=8), OVX SM TERPY (n=8) e OVX SM RIS TERPY (n=8). Em t=0, as ratas foram submetidas à cirurgia fictícia (SHAM) e à cirurgia de ovariectomia bilateral (OVX); após a recuperação da cirurgia, os animais receberam a dieta de cafeteria (SM). Passados 30 dias (t=30), o tratamento medicamentoso com risedronato de sódio (0,7 mg/kg/semana) (RIS) ou solução salina (0,3 ml), via gavagem, foi iniciado e realizado até o momento da eutanásia. Sessenta dias após o início do tratamento medicamentoso (t=90), a cirurgia de exodontia do primeiro molar superior foi realizada junto à instalação imediata dos implantes de forma bilateral (CONV ou TERPY). Quatorze dias após a cirurgia de instalação dos implantes (t=104), todos os grupos experimentais receberam a injeção intramuscular do fluorocromo calceína (20 mg/kg) e após 10 dias (t=114), do fluorocromo vermelho de alizarina (25 mg/kg). Aos 28 dias pós-operatórios (dia da eutanásia, t=118), os animais foram anestesiados e, nas maxilas do lado direito, os implantes foram submetidos ao torque reverso e imediatamente após a remoção dos implantes, foi realizada a coleta do tecido ósseo para análise de PCR tempo real para avaliação da expressão relativa de ALP, iBSP, OCN, OPG, RANKL, TRAP e VEGF, seguido da eutanásia dos animais; as maxilas do lado esquerdo foram coletadas para a análise de Micro-CT (BV/TV, Tb.Th, Tb.N, Tb.Sp e i.S) e, após o escaneamento, as mesmas passaram pelo processamento para análise da dinâmica óssea por fluorocromos (MAR e ELCOI). Os dados foram submetidos à análise estatística, com o nível de significância de 5% (p<0,05). Resultados: Fase 1: poucas diferenças foram observadas entre as duas concentrações avaliadas e assim, a menor concentração do fármaco foi selecionada (10µM). Fase 2: os implantes funcionalizados com a TERPY apresentaram os maiores valores absolutos de torque de remoção para todos os grupos e com diferença estatística para OVX SM TERPY (p=0,0402). A associação sistêmica entre o risedronato e a TERPY em ratas ovariectomizadas (OVX SM RIS TERPY) demonstrou expressão aumentada para iBSP e equilíbrio entre OPG e RANKL, corroborando com os dados obtidos para MAR (p=0,0052) e com os parâmetros de BV/TV, Tb.Th e i.S da análise microtomográfica para esse mesmo grupo. Conclusão: o desempenho clínico dos implantes funcionalizados com TERPY foi favorável, e, quando associado à administração sistêmica de risedronato de sódio, os resultados se tornam mais promissores(AU)

Objective: characterize the peri-implant bone tissue repair process in maxilla of ovariectomized rats with metabolic syndrome induced by cafeteria diet, treated with risedronate and through placement the implants functionalized with [Ru(terpy)(bdq)NO]3+ (TERY). Materials and methods: in vitro tests performed in Phase 1 evaluated the biological and physicochemical properties of the better concentration of TERPY against osteogenic responses, for the functionalization of implants using the LbL technique. In vivo experiments in Phase 2 evaluated the effect of functionalized surface during peri-implant bone repair. For this, 48 female rats were divided: SHAM CONV (n=8), OVX SM CONV (n=8), OVX SM RIS CONV (n=8), SHAM TERPY (n=8), OVX SM TERPY (n=8) and OVX SM RIS TERPY (n=8). At t=0, the rats underwent unreal surgery (SHAM) and bilateral ovariectomy surgery (OVX); after recovery from surgery, animals received cafeteria diet (SM). After 30 days (t=30), drug treatment with risedronate sodium (0.7 mg/kg/week) (RIS) or saline solution (0.3 ml), via gavage, was started and sustained out until the time of euthanasia. Sixty days after the start of drug treatment (t=90), the maxillary first molar extraction surgery was performed followed by the immediate installation of the implants bilaterally (CONV or TERPY). Fourteen days after implant placement surgery (t=104), all experimental groups received intramuscular injection of the fluorochrome calcein (20 mg/kg) and after 10 days (t=114), the red fluorochrome alizarin (25 mg/kg). After 28 postoperative days (day of euthanasia, t=118), the animals were anesthetized and, in the maxillary on the right side, the implants were subjected to reverse torque and immediately after removal of the implants, bone tissue was collected for real-time PCR analysis to measure the relative expression of ALP, iBSP, OCN, OPG, RANKL, TRAP e VEGF, followed by euthanasia of animals; the left side maxillary were collected for Micro-CT analysis (BV/TV, Tb.Th, Tb.N, Tb.Sp e i.S) and, after scanning, they underwent processing for analysis of bone dynamics by fluorochromes (MAR and ELCOI). Data values were subjected to statistical analysis, with a significance level of 5% (p<0.05). Results: Phase 1: few differences were observed between the two concentrations evaluated and thus, the lowest drug concentration was selected (10µM). Phase 2: the implants functionalized with TERPY presented the highest absolute values of removal torque for all groups and with statistical difference for OVX SM TERPY (p=0.0402). The systemic association between risedronate and TERPY in ovariectomized rats (OVX SM RIS TERPY) showed increased expression for iBSP and balance between OPG and RANKL, corroborating the data obtained for MAR (p=0.0052) and with the parameters BV/TV, Tb.Th and i.S of the microtomographic analysis for this same group. Conclusion: the clinical performance of implants functionalized with TERPY was favorable, and when associated with the systemic administration of risedronate sodium, the results become more promising(AU)

Different Stages of Alveolar Bone Repair Process Are Compromised in the Type 2 Diabetes Condition: An Experimental Study in Rats.

The aim of this study was to analyze the stages of the alveolar bone repair in type 2 diabetic rats evaluating the mechanism of mineralization and bone remodeling processes after dental extraction. Forty-eight rats were divided into normoglycemic (NG) and type 2 diabetes (T2D) groups. The upper right incisor was extracted and after 3, 7, 14 and 42 days the animals were euthanized. The following analyses were performed: immunolabeling against antibodies TNFα, TGFß, IL6, WNT, OCN and TRAP, collagen fibers maturation, microtomography and confocal microscopy. Data were submitted to statistical analysis. The immunolabeling analysis showed that the T2D presented a more pronounced alveolar inflammation than NG. Labeling of proteins responsible for bone formation and mineralization was higher in NG than T2D, which presented greater resorptive activity characterized by TRAP labeling. Also, T2D group showed a decrease in the amount of collagen fibers. Micro-CT analysis showed that T2D causes a decrease in bone volume percentage due to deficient trabecular parameters and higher porosity. The T2D bone dynamics show a loss in bone remodeling process. T2D prolongs the local inflammatory process, which impairs the organization and maturation of collagen fibers, delaying bone formation that generates impact on mineralization and bone turnover.

Sonochemical time standardization for bioactive materials used in periimplantar defects filling.

The aim of this study was determinate the best sonochemical time in order to obtain better bone characteristics when a bioactive material (Biogran) is used in the filling periimplantar defects. In this study, 32 rats were submitted to surgical proceedings to create a periimplantar defect that was filled with Biogran receiving different sonochemical times: 15 (G1), 30 (G2), 45 (G3) or 90 min (G4). The biomaterial was characterized through X-ray diffraction and scanning electron microscopy (SEM). In vivo analysis was performed through micro CT, laser confocal microscopy, immunohistochemistry and evaluation of bone cytoarchitecture through hematoxylin and eosin (HE) staining. The data were submitted to statistical testing, considering a significance level of p < 0.05. Rx diffraction of pure bioglass showed that it is predominantly amorphous; otherwise, there are small peaks at 23° and 31°. SEM shows that the longer the sonochemical time, the less edges the biomaterial will present. Within the groups, G1 and G2 showed the best quantity and quality by micro CT (p > 0.05). The best bone turnover result was found in G1 and G2, otherwise the better results were related to neoformed bone area, bone mineral apposition rate and bone implant contact to G1 (p < 0.05). G1 had the best results in terms of bone cytoarchitectural evaluation and immunohistochemistry. It is possible to conclude that Biogran that received 15 min of sonochemical treatment (G1) presented periimplantar bone repair with the best extracellular matrix properties, including the best quality and quantity of vital bone.

Losartan reverses impaired osseointegration in spontaneously hypertensive rats.

OBJECTIVE: Hypertension is not only associated with cardiovascular diseases but also with alterations in bone quality. Hypertension therefore might be a risk factor for osseointegration. Preclinical studies suggest that losartan, an angiotensin II receptor blocker widely used to treat hypertension, has a beneficial effect in graft consolidation. Here, we determine the effect of hypertension and losartan on osseointegration. METHODS: We used spontaneously hypertensive rats (SHR) and normotensive Wistar albinus rats receiving losartan (30 mg/kg, p.o.) or left untreated. After 1 week, titanium miniscrews were inserted into the tibia. Sixty days after implantation, implant stability was evaluated by removal torque measurement considered the primary endpoint. Microcomputed tomography and histomorphometric analysis were secondary endpoints. RESULTS: Losartan increased the removal torque in the hypertensive SHR group to levels of the Wistar controls. While the cortical parameters of osseointegration remained unchanged, losartan increased medullary bone formation. Microcomputed tomography revealed a higher bone volume per tissue volume and trabecular thickness in the SHR rats treated with losartan. Histomorphometric analysis further showed that losartan significantly increased the thickness of newly formed bone in medullary area in hypertensive SHR rats. Losartan did not significantly alter the parameters of osseointegration in normotensive rats. CONCLUSIONS: The data presented suggest that the angiotensin II receptor antagonist losartan increases the medullary parameters of osseointegration in a tibia model of spontaneously hypertensive rats. Considering the study limitations, understanding the impact of hypertension and the respective drugs on osseointegration requires further research.

Daily melatonin administration improves osseointegration in pinealectomized rats.

INTRODUCTION: The hypothesis of this study was that the peri-implant bone healing of the group of pinealectomized rats would differ from the control group. The samples were subjected to immunohistochemical, microtomographic (total porosity and connectivity density), and fluorochrome (mineralized surface) analyses. OBJECTIVES: The goal of this study was to investigate the cellular changes and bone remodeling dynamics along the bone/implant interface in pinealectomized rats. MATERIAL AND METHODS: The total of 18 adult male rats (Rattus norvegicus albinus, Wistar) was divided into three groups (n=6): control (CO), pinealectomized without melatonin (PNX) and pinealectomized with melatonin (PNXm). All animals were submitted to the first surgery (pinealectomy), except the CO group. Thirty days after the pinealectomy without melatonin, the second surgery was conducted, in which all animals received an implant in each tibia (36 titanium implants with surface treatment were installed - Implalife® São Paulo, SP, Brazil). By gavage, the rats of the PNX group received the vehicle solution, and the procedure. RESULTS: Immunohistochemical analysis for runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteopontin (OP) and osteocalcin (OC) showed that the bone repair process in the PNXm group was similar to that of the CO group, whereas the PNX group showed a delay. The microtomographic parameters of total porosity [Po(tot)] and bone surface (BS) showed no statistically significant differences, whereas for the connective density (Conn.Dn) a statistical difference was found between the CO and PNXm groups. Fluorochrome analysis of the active mineralized surface showed statistically significant difference between the CO and PNX and between the CO and PNXm groups. CONCLUSION: The absence of the pineal gland impaired the bone repair process during osseointegration, however the daily melatonin replacement was able to restore this response.

Comparative Evaluation of Cell Viability Immediately After Osteotomy for Implants With Drills and Piezosurgery: Immunohistochemistry Analysis.

OBJECTIVE: To evaluate the effect of reusing drills and piezosurgery tips during implant osteotomy on immediate bone cell viability through immunohistochemical analysis. MATERIALS AND METHODS: Six male rabbits were divided into 2 groups and then divided into 5 subgroups-correspond to drills and tips used 10, 20, 30, 40, and 50 times, respectively. All animals received 10 osteotomies in each tibia, by use of the classic drilling procedure in one group (G1) and the piezosurgery device in the other group (G2). For immunohistochemical technique were utilized the osteoprotegerin, RANKL, osteocalcin, and caspase 3. Control procedures were performed by omitting the primary antibodies (negative control). RESULTS: Bone formation and resorption responses presented in more intense way during the piezosurgery. The expression of osteocalcin had become quite intense in piezosurgery groups, but with reduced immunostaining from the 30th osteotomy. The caspase 3 showed the viability of the osteoblast from the 20th osteotomy with piezosurgery and remained constant until the 50th. CONCLUSION: Piezosurgery provides greater osteoblastic cell viability than the system of conventional drilling. CLINICAL RELEVANCE: This study will provide data so that the authors can recycle the drills and tips for implant placement, thus enabling a better cell viability for osseointegration.

Bone repair with raloxifene and bioglass nanoceramic composite in animal experiment.

OBJECTIVE: This study proposes to evaluate, by means of microtomographic analysis, the topical potential of raloxifene combined with BioGran® through the sonochemical method in the repair of critical bone defects in the calvaria of rats. The hypothesis was that the homogenization of Raloxifene to Biogran at the 20% concentration would improve the bone repair at the grafted site. MATERIALS AND METHODS: A 5-mm calvaria bone defect was induced in three groups: CTR (100% BioGran®); RAL10 (90% BioGran® and 10% raloxifene), and RAL20 (80% BioGran® and 20% raloxifene). The animals were euthanized after 30 days and the microCT analysis was then performed to evaluate the parameters bone volume (BV), bone volume percentage (BV/TV), trabecular bone thickness (Tb.Th), and the separation and number of trabeculae (Tb.Sp and Tb.N). The obtained results were compared using ANOVA and Tukey test (p < 0.05). RESULTS: The best results were found for the CTR and RAL20 groups, in which the BV, BV/TV, Tb.Sp, and Tb.N parameters were statistically significant in comparison with RAL10 (p < 0.05). CONCLUSIONS: In view of the results obtained in this experiment, we can conclude that BioGran® alone or in an 80/20 mass concentration with raloxifene can lead to favorable bone formation.

Delayed Diagnosis of Gorlin-Goltz Syndrome: The Importance of the Multidisciplinary Approach.

Gorlin-Goltz syndrome (GGS), also known as nevoid basal cell carcinoma syndrome, is an autosomal dominant inherited disorder with high penetrance and variable expressivity. The classic triad originally described by Gorlin and Goltz in 1960 is composed of multiple nevoid basal cell carcinomas (NBCCs), odontogenic keratocysts (OKCs) in the jaws and bifid ribs. in 1977, this triad was modified by Rayner et al, and to GGS diagnosis, the OKCs had to appear in combination with calcification of the cerebellar falx or palmar and plantar pits. It may occur that although GGS syndrome is a well-known condition, only the specific symptom could be observed by different specialists. Therefore, the patient cannot be placed in an always complex clinical panel. The authors introduce an example in this report. In the present case, the patient had NBCCs, OKCs, and probably other signs of GGS since 1998, and has been treated for this conditions separated, without a diagnosis of a syndromic condition. A 54-year-old white woman was referred to the oral medicine service due to cyst located in the right mandibular body. She had history of skin cancer and undergone surgeries and radiotherapies for the lesions treatment, scars on the skin face due to the lesions removed, and a new ulcerated lesion on the back of was diagnosed. In addition, the patient presented frontal and parietal bossing leading to increased cranial circumference, hypertelorism, strabismus, broad base, and mandibular prognathism. To the image examination, skull radiography revealed calcification of the falx cerebri; on chest X-ray bifid rib was observed and spine radiography showed vertebral osteophytes. Panoramic radiograph showed a well-defined bilocular radiolucent image located in posterior and anterior mandibular region. The whole elements induced us to investigate the patient's past medical history, which revealed that since 1998 had the diagnosis of NBCC and OKC. A multidisciplinary approach becomes necessary for the diagnosis and follow-up of patients with GGS, considering the complexity of the clinical manifestations. Therefore, it is of primary importance for dental surgeons and dermatologists to know the signs and symptoms of GGS to perform early diagnosis and to avoid progression of the oral cysts or metastasis of the skin lesions.

Avaliação do reparo ósseo na interface osso/implante em ratas com deficiência de estrógeno tratadas com fitoestrógeno genisteína: análise microtomográfica, molecular por PCR e imunoistoquímica / Evaluation of bone repair at the bone / implant interface in estrogen deficient rats treated with phytoestrogen genistein: microtomographic, molecular analysis by PCR and immunohistochemistry

O presente projeto teve como proposta avaliar o processo de reparo ósseo na interface osso/implante em ratas submetidas à ovariectomia para a indução da deficiência de estrógeno e tratadas com fitoestrógeno genisteína (1mg/dia) por gavagem. Para isso, 30 ratas (Rattus novergicus albinus, Wistar), com peso médio de 200 gramas foram divididas em 3 grupos conforme a análise a que foram submetidas e o tratamento medicamentoso, sendo 10 ratas para o grupo SHAM SAL = Ratas submetidas à cirurgia fictícia, sem tratamento medicamentoso (gavagem com solução salina 0.9%); 10 ratas para o grupo OVX SAL = Ratas submetidas à ovariectomia bilateral, sem tratamento medicamentoso (gavagem com solução salina 0.9%); 10 ratas para o grupo OVX GEN = Ratas submetidas à ovariectomia bilateral, tratadas com fitoestrógeno genisteína (gavagem com 1mg/dia de genisteína). Foram utilizados 60 implantes, cada animal recebeu 2 implantes, sendo 1 em cada metáfise tibial. A eutanásia dos animais foi realizada após 60 dias da instalação dos implantes, por meio de sobredosagem anestésica. Para os grupo calcificados: nas anális es por microtomografia computadorizada foram utilizadas as tíbias do lado direito de 6 ratas e para a análise da expressão gênica por PCR em tempo real, as 6 tíbias do lado esquerdo das mesmas ratas. Para a análise dos grupos descalcificados: 4 ratas foram submetidas à eutanásia na análise de imunoistoquímica osteoprotegerina ­ (OPG-SC21038), ativador do receptor nuclear kappa B ligante ­ (RANKL-SC 7627), osteocalcina ­ (OC-SC18319) e fosfatase ácida resistente ao ácido tartárico (TRAP-SC30832) e fosfatase alcalina (ALP -SC23430). Todos os dados quantitativos foram submetidos ao teste de homocedasticidade e após a determinação do teste mais adequado, o nível de significância foi estabelecido em 0<0,05. A análise morfométrica do grupo OVX GEN mostrou maior percentual de volume ósseo, menor separação entre as trabéculas e menor porosidade. As análises molecular por PCR e Imunohistoquímica evidenciam uma maior expressão para OPG e RANKL (remodelação óssea), ALP e OC (mineralização) de cada um desses marcadores, o que sugere que a genisteína atua como um modulador seletivo natural dos receptores de estrogenio favorecendo a expressão de proteínas importantes para a modulação da atividade osteoblástica no metabolismo do tecido ósseo. Assim, os resultados deste estudo confirmaram que a terapia com genisteína mostra-se capaz de melhorar a reparação óssea na interface osso / implante sob condições de osteopenia/osteoporose(AU)

The present project had as proposal to evaluate the process of bone repair in the bone / implant interface in rats submitted to ovariectomy for induction of estrogen deficiency and treated with phytoestrogen genistein (1mg / day) by gavage. For this, 30 rats (Rattus novergicus albinus, Wistar), with an average weight of 200 grams were divided into 3 groups according to an analysis and the drug treatment, being 10 rats for the group SHAM SAL = Rats submitted to the fictional surgery , without drug treatment (gavage with 0.9% saline solution); 10 rats for the OVX group SAL = Rats submitted to bilateral ovariectomy without drug treatment (gavage with 0.9% saline solution); 10 rats for the OVX group GEN = Rats subjected to bilateral ovariectomy, treated with phytoestrogen genistein (gavage with 1mg / day of genistein). 60 implants were installed, with 2 implants in each animal. The euthanasia of the animals was performed 60 days after implant's installation, by anesthetic overdose. The calcified groups analysis with computerized microtomography was peformed in right tibiae of 6 rats and the gene expression analysis by PCR was performed in left tíbia of the same animals. For the decalcified analysis through immunolabeling experiments, 4 rats were submited to eutanásia in order to evaluate the presence of Osteoprotegerin (OPG ­ SC21038), activator of nuclear kappa B ligand (RANKL ­ SC7627), Osteocalcin (OC ­ SC 18319), and acid resistant phosphatase tartaric acid (TRAP ­ SC 30832) and alcaline phosphatase (ALP SC 23430). All the quantitative data were submitted to the normality curve to determine the most adequate test. The significance level of p<0.05 was considered for all tests. A morphometric analysis of the OVX GEN group showed higher percentage of bone volume, less separation between trabeculae and lower porosity. As molecular analyzes by PCR and Immunohistochemistry evidenced a greater expression for OPG and RANKL (bone remodeling), ALP and OC (mineralization) of each the markers, which suggests that genistein acts as a natural selective modulator of estrogen receptors favoring expression of proteins important for a modulation of osteoblastic activity in the metabolism of bone tissue. Thus, the results of this study confirm that genistein therapy is shown to improve bone repair at the bone / implant interface under conditions of osteopenia / osteoporosis(AU)

Daily melatonin administration improves osseointegration in pinealectomized rats

Abstract The hypothesis of this study was that the peri-implant bone healing of the group of pinealectomized rats would differ from the control group. The samples were subjected to immunohistochemical, microtomographic (total porosity and connectivity density), and fluorochrome (mineralized surface) analyses. Objectives The goal of this study was to investigate the cellular changes and bone remodeling dynamics along the bone/implant interface in pinealectomized rats. Material and Methods The total of 18 adult male rats (Rattus norvegicus albinus, Wistar) was divided into three groups (n=6): control (CO), pinealectomized without melatonin (PNX) and pinealectomized with melatonin (PNXm). All animals were submitted to the first surgery (pinealectomy), except the CO group. Thirty days after the pinealectomy without melatonin, the second surgery was conducted, in which all animals received an implant in each tibia (36 titanium implants with surface treatment were installed - Implalife® São Paulo, SP, Brazil). By gavage, the rats of the PNX group received the vehicle solution, and the procedure. Results Immunohistochemical analysis for runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteopontin (OP) and osteocalcin (OC) showed that the bone repair process in the PNXm group was similar to that of the CO group, whereas the PNX group showed a delay. The microtomographic parameters of total porosity [Po(tot)] and bone surface (BS) showed no statistically significant differences, whereas for the connective density (Conn.Dn) a statistical difference was found between the CO and PNXm groups. Fluorochrome analysis of the active mineralized surface showed statistically significant difference between the CO and PNX and between the CO and PNXm groups. Conclusion The absence of the pineal gland impaired the bone repair process during osseointegration, however the daily melatonin replacement was able to restore this response.