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.

Avaliação do efeito sinérgico entre risedronato sistêmico e genisteína local sobre o reparo peri-implantar em ratas com deficiência de estrógeno e síndrome metabólica / Evaluation of the synergistic effect between systemic Risedronate and local Genistein on peri-implant repair in strogen deficient rats and metabolic syndrome

O objetivo deste estudo foi investigar a ação sinérgica do risedronato de sódio sistêmico e da genisteína administrada localmente, através da funcionalização de implantes, de ratas submetidas a ovariectomia e com hábitos de mimetizam a síndrome metabólica. A parte in vitro deste estudo foi executado em 2 etapas. Na primeira etapa, foi realizada a funcionalização da superfície de discos/implantes com genistína na concentração de 100 µM pela técnica layer by layer (lbl). Na segunda etapa foram feitos testes biológicos em culturas de células, para avaliar as propriedades da superfície funcionalizada, quanto às respostas osteogênicas. Para a cultura de células foram utilizadas células mesenquimais diferenciadas em osteoblastos, isoladas de fêmures de ratos. Após a validação pelos testes executados nas superfícies funcionalizadas, foi realizado estudo in vivo (3ª etapa). Para tanto, no dia 0, as ratas Wistar adultas jovens, fêmeas (n=64) foram divididas em 4 grupos: 1- SHAM (n= 16), animais foram submetidos à ovariectomia (OVX) fictícia e dieta balanceada. 2- SHAM Síndrome Metabólica (SM) (n=16), animais foram submetidos à ovariectomia fictícia e dieta de cafeteria. 3- OVX SM (n=16), animais foram submetidos à ovariectomia bilateral e dieta de cafeteria. 4- OVX SM Risedronato (RIS) (n=16), animais foram submetidos à ovariectomia bilateral, dieta de cafeteria e tratadas com risedronato de sódio. Em cada grupo há 2 subgrupos: A- implantes convencionais e B- implantes funcionalizados com genisteína. No dia 30, foi iniciado o tratamento medicamentoso com risedronato de sódio, na concentração de 0,35mg/kg, ou apenas solução salina, via gavagem, 1 vez por semana. Passados 60 dias da medicação (dia 90), todos os animais foram submetidos à cirurgia para exodontia dos 1os molares superiores bilateralmente e, imediatamente, no alvéolo da raiz mesial, foi instalado os implantes com superfície convencional ou funcionalizada. Os animais foram eutanasiados aos 28 dias (dia 118) após a instalação dos implantes para mensuração do torque de falha na interface osso implante em N/cm. Os dados foram submetidos ao teste de homocedasticidade (Shapiro Wilk). Houve a confirmação de distribuição normal dos dados amostrais e na sequência, foi realizado o teste paramétrico ANOVA One Way or Two Way, seguido do pós teste de Tukey, com o nível de significância de 5% (p< 0,05). Concluiu-se que, a concentração de 100 µM da genisteína manteve a viabilidade celular e resultados favoráveis quanto a genotoxicidade. A dieta de cafeteria e a ovariectomia bilateral mimetizam a síndrome metabólica e a predisposição para osteoporose por deficiência de esteroides gonadais. E, a ação sinérgica entre fármaco sistêmico (risedronato de sódio) e genisteína local foi promissora para a melhora no processo de reparo periimplantar, principalmente no grupo SHAM e OVX SM RIS(AU)

The aim of this study was to investigate the synergistic action of systemic risedronate sodium and locally administered genistein, through implant functionalization, of rats submitted to ovariectomy and with habits mimicking the metabolic syndrome. The in vitro part of this study was performed in 2 steps. In the first step, the surface functionalization of discs/implants was performed with genistein at a concentration of 100 µM by the layer by layer (lbl) technique. In the second step biological tests were performed in cell cultures to evaluate the properties of the functionalized surface for osteogenic responses. For the cell culture, mesenchymal cells differentiated into osteoblasts, isolated from rat femurs, were used. After validation by tests performed on the functionalized surfaces, the in vivo study (third test) was performed. For this purpose, on day 0, young adult female Wistar rats (n=64) were divided into 4 groups: 1- SHAM (n=16), animals were submitted to sham ovariectomy (OVX) and balanced diet. 2- SHAM Metabolic Syndrome (MS) (n=16), animals were submitted to sham ovariectomy and cafeteria diet. 3- OVX SM (n=16), animals underwent bilateral ovariectomy and cafeteria diet. 4- OVX SM Risedronate (RIS) (n=16), animals underwent bilateral ovariectomy, cafeteria diet and treated with risedronate sodium. In each group there are 2 subgroups: A- conventional implants and B- implants functionalized with genistein. On day 30, drug treatment was started with risedronate sodium, at a concentration of 0.35 mg/kg, or just saline solution, via gavage, once a week. After 60 days of medication (day 90), all animals underwent surgery to extract the 1st upper molars bilaterally, and implants with conventional or functionalized surfaces were immediately installed in the mesial root alveolus. The animals were euthanized at 28 days (day 118) after implant installation to measure the failure torque at the implant-bone interface in N/cm. The data were submitted to the homoscedasticity test (Shapiro Wilk). The normal distribution of the sample data was confirmed and then the parametric One Way or Two Way ANOVA test was performed, followed by Tukey's post-test, with a significance level of 5% (p< 0.05). It was concluded that, the concentration of 100 µM of genistein maintained cell viability and favorable results regarding genotoxicity. The cafeteria diet and bilateral ovariectomy mimic the metabolic syndrome and predisposition to osteoporosis by gonadal steroid deficiency. And, the synergistic action between systemic drug (risedronate sodium) and local genistein was promising for the improvement in the periimplant repair process, especially in the SHAM and OVX SM RIS groups(AU)

Gene expression, immunohistochemical and microarchitectural evaluation of bone formation around two implant surfaces placed in bone defects filled or not with bone substitute material.

OBJECTIVE: The aim of this study is to evaluate through gene expression, immunohistochemical and microtomographic (micro-CT) analysis the response of peri-implant bone tissue around titanium implants with different surface treatments, placed in bone defects filled or not with bone substitute materials. In addition, to investigate the hypothesis that porous-hydrophilic surface induces a faster bone formation. MATERIALS AND METHODS: Twenty-six animals were divided into two groups according to implant surface treatment. In each tibia, a bone defect was created followed by the placement of one implant. On the left tibia, the defect was filled with blood clot (BC), and on the right tibia, the defect was filled with biphasic hydroxyapatite/ß-tricalcium-phosphate (HA/TCP) generating four subgroups: BC-N: bone defect filled with blood clot and porous surface titanium implant installed; BC-A: bone defect filled with blood clot and porous-hydrophilic surface titanium implant installed; HA/TCP-N: bone defect filled with bone substitute material and porous surface titanium implant installed; and HA/TCP-A: bone defect filled with bone substitute material and porous-hydrophilic surface titanium implant installed. The animals were submitted to euthanasia at 15, 30, and 60 days after implant installation. The expression of two genes was evaluated: RUNX2 and BSP. Immunohistochemical analyses were performed for detection of RUNX2, OPN, OCN, OPG, and RANKL antibodies and bone matrix proteins. Finally, four parameters were chosen for micro-CT analysis: trabecular number, separation and thickness, and connectivity density. RESULTS: Descriptive analysis showed similar findings among the experimental groups. Moreover, porous-hydrophilic surfaces presented a higher expression of RUNX2, which is probably an indicative of better osteogenesis; although the data from this study may be considered an insufficient support for a concrete statement. CONCLUSION: Porous hydrophilic surface can improve and accelerate protein expression and bone formation.

Author Correction: Plasma Electrolytic Oxidation as a Feasible Surface Treatment for Biomedical Applications: an in vivo study.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Plasma Electrolytic Oxidation as a Feasible Surface Treatment for Biomedical Applications: an in vivo study.

OBJECTIVES: In this in vivo animal study, we evaluated the effect of plasma electrolytic oxidation (PEO) coating on the topographic and biological parameters of implants installed in rats with induced osteoporosis and low-quality bones. MATERIALS AND METHODS: In total 44 Wistar rats (Rattus novergicus), 6 months old, were submitted to ovariectomy (OXV group) and dummy surgery (SHAM group). After 90 days, the ELISA test was performed and the ovariectomy effectiveness was confirmed. In each tibial metaphysis, an implant with PEO coating containing Ca2+ and P5+ molecules were installed, and the other tibia received an implant with SLA acid etching and blasting (AC) (control surface). After 42 days, 16 rats from each group were euthanized, their tibias were removed for histological and immunohistochemical analysis (OPG, RANKL, OC and TRAP), as well as reverse torque biomechanics. Data were submitted to One-way ANOVA or Kruskal-Wallis tests, followed by a Tukey post-test; P < 0.05. Histological analyses showed higher bone neoformation values among the members of the PEO group, SHAM and OVX groups. Immunohistochemical analysis demonstrated equilibrium in all groups when comparing surfaces for TRAP, OC and RANKL (P > 0.05), whereas OPG showed higher PEO labeling in the OVX group (P < 0.05). Biomechanical analysis showed higher reverse torque values (N.cm) for PEO, irrespective of whether they were OVX or SHAM groups (P < 0.05). CONCLUSION: The results indicated that the PEO texturing method favored bone formation and showed higher bone maturation levels during later periods in osteoporotic rats.

Teriparatide improves alveolar bone modelling after tooth extraction in orchiectomized rats.

OBJECTIVE: The aim of this study was to evaluate bone metabolism in the alveolar repair process in orchiectomized male rats treated with teriparatide. MATERIALS AND METHODS: A total of 78 rats were divided into three groups: 26 oriectomized rats treated with teriparatide (ORQTRAT), 26 untreated orchiectomized rats (ORQ), and 26 rats which had undergone sham surgical procedures (SHAM), all these animals underwent extraction of the central incisor tooth. Thus, a histological analysis was performed (42 days). Real Time PCR (14 and 42 days) and immunohistochemical (42 days) analyses were performed based on the expression of RANK-L and osteoprotegerin (OPG). The calcein and alizarin were analyzed via laser confocal microscopy to verify alveolar bone turnover, and microtomographic analysis was performed to determine volume and bone quality (calcified tissues). In this analysis, the euthanasia period was 60 days post-extraction. The quantitative data were submitted to the statistical test and the significance level of 5% was adopted. RESULTS: Teriparatide increased bone turnover with a higher relative gene expression of RANKL and OPG at 14 days, and at 42 days, there was a significant decrease of RANKL and an increase of OPG, this standard can also be evaluated in the ratio of RANKL to OPG. Greater values of area and bone mineral deposition were found for the ORQTRAT group (p < 0.05), along with higher bone volume values (p < 0.05). The immunolabels revealed greater intensity in the relationship of RANKL and OPG, which led to intense remodeling aiding in the process of bone formation. CONCLUSION: The teriparatide treatment in orchiectomized rats increases bone volume and decreases the porosity, in addition to promoting greater intensity in bone turnover during alveolar repair.

Alveolar bone healing in rats: micro-CT, immunohistochemical and molecular analysis.

INTRODUCTION: Alveolar bone healing after upper incisor extraction in rats is a classical model of preclinical studies. The underlying morphometric, cellular and molecular mechanism, however, remains imprecise in a unique study. OBJECTIVES: The aim of this study was therefore to characterize the alveolar bone healing after upper incisor extraction in rats by micro computed tomographic (Micro-CT), immunohistochemical and real-time polymerase chain reaction (RT-PCR) analysis. MATERIAL AND METHODS: Thirty animals (Rattus norvegicus, Albinus Wistar) were divided into three groups after upper incisors extraction at 7, 14, and 28 days. Micro-CT was evaluated based on the morphometric parameters. Subsequently, the histological analyses and immunostaining of osteoprotegerin (OPG), receptor activator of nuclear kappa B ligand (RANKL) and tartrate resistant acid phosphate (TRAP) was performed. In addition, RT-PCR analyses of OPG, RANKL, the runt-related transcription factor 2 (RUNX2), osteocalcin (OC), osteopontin (OPN), osterix (OST) and receptor activator of nuclear kappa B (RANK) were performed to determine the expression of these proteins in the alveolar bone healing. RESULTS: Micro-CT: The morphometric parameters of bone volume and trabecular thickness progressively increased over time. Consequently, a gradual decrease in trabecular separation, trabecular space and total bone porosity was observed. Immunohistochemical: There were no differences statistically significant between the positive labeling for OPG, RANKL and TRAP in the different periods. RT-PCR: At 28 days, there was a significant increase in OPG expression, while RANKL expression and the RANKL/OPG ratio both decreased over time. CONCLUSION: Micro-CT showed the newly formed bone had favorable morphometric characteristics of quality and quantity. Beyond the RUNX2, OC, OPN, OST, and RANK proteins expressed in the alveolar bone healing, OPG and RANKL activity showed to be essential for activation of basic multicellular units during the alveolar bone healing.

A SERM increasing the expression of the osteoblastogenesis and mineralization-related proteins and improving quality of bone tissue in an experimental model of osteoporosis.

Raloxifene is an antiresorptive drug, selective estrogen receptor modulator (SERM) used in the treatment of osteoporosis. Objective To evaluate proteins related to bone repair at the peri-implant bone in a rat model of osteoporosis treated with raloxifene. Material and Methods 72 rats were divided into three groups: SHAM (healthy animals), OVX (ovariectomized animals), and RLX (ovariectomized animals treated with raloxifene). Raloxifene was administered by gavage (1 mg/kg/day). Tibial implantation was performed 30 days after ovariectomy, and animals were euthanized at 14, 42, and 60 days postoperatively. Samples were collected and analyzed by immunohistochemical reactions, molecular analysis, and microtomographic parameters. Results RLX showed intense staining of all investigated proteins at both time points except for RUNX2. These results were similar to SHAM and opposite to OVX, showing mild staining. The PCR gene expression of OC and ALP values for RLX (P<0.05) followed by SHAM and OVX groups. For BSP data, the highest expression was observed in the RLX groups and the lowest expression was observed in the OVX groups (P<0.05). For RUNX2 data, RLX and SHAM groups showed greater values compared to OVX (P<0.05). At 60 days postoperatively, microtomography parameters, related to closed porosity, showed higher values for (Po.N), (Po.V), and (Po) in RLX and SHAM groups, whereas OVX groups showed lower results (P<0.05); (BV) values (P=0.009); regarding total porosity (Po.tot), RLX group had statistically significant lower values than OVX and SHAM groups (P=0.009). Regarding the open porosity (Po.V and Po), the SHAM group presented the highest values, followed by OVX and RLX groups (P<0.05). The Structural Model Index (SMI), RLX group showed a value closer to zero than SHAM group (P<0.05). Conclusions Raloxifene had a positive effect on the expression of osteoblastogenesis/mineralization-related proteins and on micro-CT parameters related to peri-implant bone healing.

Raloxifene but not alendronate can compensate the impaired osseointegration in osteoporotic rats.

OBJECTIVES: Alendronate and raloxifene, a bisphosphonate and a selective estrogen modulator, respectively, are established osteoporosis therapies. Current evidence suggests that simultaneous application of osteoporosis therapies modulates osseointegration. However, alendronate shows inconsistent findings and raloxifene has not been studied comprehensively. This study aimed to evaluate the bone dynamics and molecular and microstructural features at the peri-implant bone interface in osteoporotic rats. MATERIALS AND METHODS: Thirty female rats underwent ovariectomy and were fed a diet low in calcium and phosphate and treated with alendronate or raloxifene for 30 days or underwent fictional ovariectomy surgery (SHAM) prior to implant insertion in the tibia; osteoporosis therapies continued thereafter. After 42 days, peri-implant bone was evaluated by histometric and micro-CT analysis. Fluorochrome incorporation and gene expression was determined to evaluate bone turnover. RESULTS: We report here that alendronate had no impact on bone-to-implant contacts and the mineral apposition rate. The RANKL/OPG ratio and local bone volume, however, were increased compared to the untreated osteoporotic rats. Even though signaling to bone resorption activity through RANKL production was observed in the alendronate group, the blockade of bone resorption activity that occurs in decorrence to alendronate activity took place and resulted in an increase in bone volume. Raloxifene significantly increased osseointegration in osteoporotic rats, as indicated by bone-to-implant contacts, mineral apposition, and local bone volume. Raloxifene, however, had no considerable impact on the RANKL/OPG ratio compared to untreated osteoporotic rats. As expected, the SH group showed higher bone-to-implant contacts and mineral apposition rates than the untreated osteoporotic rats. CONCLUSIONS: These findings suggest that raloxifene but not alendronate can compensate for the impaired osseointegration in osteoporotic rats. CLINICAL RELEVANCE: Regarding the superiority of raloxifene observed in the improvement of bone dynamics response, this statement suggests that raloxifene could be a good option for osteoporosis patients in oral rehabilitation procedures.

Alveolar bone healing in rats: micro-CT, immunohistochemical and molecular analysis

Abstract Alveolar bone healing after upper incisor extraction in rats is a classical model of preclinical studies. The underlying morphometric, cellular and molecular mechanism, however, remains imprecise in a unique study. Objectives The aim of this study was therefore to characterize the alveolar bone healing after upper incisor extraction in rats by micro computed tomographic (Micro-CT), immunohistochemical and real-time polymerase chain reaction (RT-PCR) analysis. Material and Methods Thirty animals (Rattus norvegicus, Albinus Wistar) were divided into three groups after upper incisors extraction at 7, 14, and 28 days. Micro-CT was evaluated based on the morphometric parameters. Subsequently, the histological analyses and immunostaining of osteoprotegerin (OPG), receptor activator of nuclear kappa B ligand (RANKL) and tartrate resistant acid phosphate (TRAP) was performed. In addition, RT-PCR analyses of OPG, RANKL, the runt-related transcription factor 2 (RUNX2), osteocalcin (OC), osteopontin (OPN), osterix (OST) and receptor activator of nuclear kappa B (RANK) were performed to determine the expression of these proteins in the alveolar bone healing. Results Micro-CT: The morphometric parameters of bone volume and trabecular thickness progressively increased over time. Consequently, a gradual decrease in trabecular separation, trabecular space and total bone porosity was observed. Immunohistochemical: There were no differences statistically significant between the positive labeling for OPG, RANKL and TRAP in the different periods. RT-PCR: At 28 days, there was a significant increase in OPG expression, while RANKL expression and the RANKL/OPG ratio both decreased over time. Conclusion Micro-CT showed the newly formed bone had favorable morphometric characteristics of quality and quantity. Beyond the RUNX2, OC, OPN, OST, and RANK proteins expressed in the alveolar bone healing, OPG and RANKL activity showed to be essential for activation of basic multicellular units during the alveolar bone healing.

A SERM increasing the expression of the osteoblastogenesis and mineralization-related proteins and improving quality of bone tissue in an experimental model of osteoporosis

Abstract Raloxifene is an antiresorptive drug, selective estrogen receptor modulator (SERM) used in the treatment of osteoporosis. Objective To evaluate proteins related to bone repair at the peri-implant bone in a rat model of osteoporosis treated with raloxifene. Material and Methods 72 rats were divided into three groups: SHAM (healthy animals), OVX (ovariectomized animals), and RLX (ovariectomized animals treated with raloxifene). Raloxifene was administered by gavage (1 mg/kg/day). Tibial implantation was performed 30 days after ovariectomy, and animals were euthanized at 14, 42, and 60 days postoperatively. Samples were collected and analyzed by immunohistochemical reactions, molecular analysis, and microtomographic parameters. Results RLX showed intense staining of all investigated proteins at both time points except for RUNX2. These results were similar to SHAM and opposite to OVX, showing mild staining. The PCR gene expression of OC and ALP values for RLX (P<0.05) followed by SHAM and OVX groups. For BSP data, the highest expression was observed in the RLX groups and the lowest expression was observed in the OVX groups (P<0.05). For RUNX2 data, RLX and SHAM groups showed greater values compared to OVX (P<0.05). At 60 days postoperatively, microtomography parameters, related to closed porosity, showed higher values for (Po.N), (Po.V), and (Po) in RLX and SHAM groups, whereas OVX groups showed lower results (P<0.05); (BV) values (P=0.009); regarding total porosity (Po.tot), RLX group had statistically significant lower values than OVX and SHAM groups (P=0.009). Regarding the open porosity (Po.V and Po), the SHAM group presented the highest values, followed by OVX and RLX groups (P<0.05). The Structural Model Index (SMI), RLX group showed a value closer to zero than SHAM group (P<0.05). Conclusions Raloxifene had a positive effect on the expression of osteoblastogenesis/mineralization-related proteins and on micro-CT parameters related to peri-implant bone healing.

Morphometric and histologic characterization of alveolar bone from hypertensive patients.

BACKGROUND: Hypertension is considered a risk factor in implant dentistry but the underlying reasons remain unclear. It may be that hypertension has a negative impact on the local bone quality. PURPOSE: Here we evaluated the structural and histological parameters of bone collected from hypertensive patients treated by antagonists of the renin-angiotensin system, and of bone collected from normotensive patients. MATERIAL AND METHODS: A total of 30 patients were referred for rehabilitation with dental implants to be placed in the posterior mandible. The first group were hypertensive patients treated with RAS antagonists. The second group were normotensive patients not taking medication. Bone biopsies collected during implant installation were subjected to analysis. Micro CT revealed the structural parameters. Histological analyses together with immunohistochemical staining of osteogenic markers were performed. RESULTS: The structural parameters of bone volume, trabecular thickness, trabecular number, separation of the trabecular, and total porosity were similar between the 2 groups (P > .05). The histological appearance of bone derived from hypertensive patients was normal. The staining pattern of Runx-2, osteopontin, and osteocalcin were comparable in both groups. CONCLUSIONS: These observations suggest that hypertensive patients treated with renin-angiotensin system antagonists have regular alveolar bone with respect to bone structure and histological parameters.

Successful Use of Lower-Level Laser Therapy in the Treatment of Medication-Related Osteonecrosis of the Jaw.

Case Presentation: This study aimed to report a case of medication related osteonecrosis of the jaw (MRONJ) of a 65-year-old female patient referred to the Oral and Maxillofacial Surgery team from Araçatuba Dental School, complaining about mobility of a previously dental implant placed on the posterior maxillary region. Clinical examination revealed an extensive necrosis area around the implant region. The patient reported bisphosphonate therapy with sodium alendronate for prevention of osteoporosis 5 years ago. A diagnosis of MRONJ was reached and the treatment decided was to remove the dental implant damaged and use the lower-level laser therapy (LLLT) associated with antibiotic therapy with clindamycin 300 mg and mouth rinses with chlorhexidine 0.12%. Conclusion: Six months after the treatment with LLLT a complete healing of the affected area was observed and 12 months after treatment the patient was rehabilitated with fixed dental prosthesis and showed excellent tissue healing of the necrosis areas. It was concluded that the use of LLLT showed to be a good option in the treatment of MRONJ.

Short term sodium alendronate administration improves the peri-implant bone quality in osteoporotic animals.

OBJECTIVE: The aim of this study was to evaluate the bone repair process at the bone/implant interface of osteoporotic rats treated with sodium alendronate through the analysis of microtomography, real time polymerase chain reactions and immunohistochemistry (RUNX2 protein, bone sialoprotein (BSP), alkaline phosphatase, osteopontin and osteocalcin). MATERIAL AND METHODS: A total of 42 rats were used and divided in to the following experimental groups: CTL: control group (rats submitted to fictitious surgery and fed with a balanced diet), OST: osteoporosis group (rats submitted to a bilateral ovariectomy and fed with a low calcium diet) and ALE: alendronate group (rats submitted to a bilateral ovariectomy, fed with a low calcium diet and treated with sodium alendronate). A surface treated implant was installed in both tibial metaphyses of each rat. Euthanasia of the animals was conducted at 14 (immunhostochemistry) and 42 days (immunohistochemistry, micro CT and PCR). Data were subjected to statistical analysis with a 5% significance level. RESULTS: Bone volume (BV) and total pore volume were higher for ALE group (P<0.05). Molecular data for RUNX2 and BSP proteins were significantly expressed in the ALE group (P<0.05), in comparison with the other groups. ALP expression was higher in the CTL group (P<0.05). The immunostaining for RUNX2 and osteopontin was positive in the osteoblastic lineage cells of neoformed bone for the CTL and ALE groups in both periods (14 and 42 days). Alkaline phosphatase presented a lower staining area in the OST group compared to the CTL in both periods and the ALE at 42 days. CONCLUSION: There was a decrease of osteocalcin precipitation at 42 days for the ALE and OST groups. Therefore, treatment with short-term sodium alendronate improved bone repair around the implants installed in the tibia of osteoporotic rats.

Short term sodium alendronate administration improves the peri-implant bone quality in osteoporotic animals

Abstract Sodium alendronate is a bisphosphonate drug that exerts antiresorptive action and is used to treat osteoporosis. Objective The aim of this study was to evaluate the bone repair process at the bone/implant interface of osteoporotic rats treated with sodium alendronate through the analysis of microtomography, real time polymerase chain reactions and immunohistochemistry (RUNX2 protein, bone sialoprotein (BSP), alkaline phosphatase, osteopontin and osteocalcin). Material and Methods A total of 42 rats were used and divided in to the following experimental groups: CTL: control group (rats submitted to fictitious surgery and fed with a balanced diet), OST: osteoporosis group (rats submitted to a bilateral ovariectomy and fed with a low calcium diet) and ALE: alendronate group (rats submitted to a bilateral ovariectomy, fed with a low calcium diet and treated with sodium alendronate). A surface treated implant was installed in both tibial metaphyses of each rat. Euthanasia of the animals was conducted at 14 (immunhostochemistry) and 42 days (immunohistochemistry, micro CT and PCR). Data were subjected to statistical analysis with a 5% significance level. Results Bone volume (BV) and total pore volume were higher for ALE group (P<0.05). Molecular data for RUNX2 and BSP proteins were significantly expressed in the ALE group (P<0.05), in comparison with the other groups. ALP expression was higher in the CTL group (P<0.05). The immunostaining for RUNX2 and osteopontin was positive in the osteoblastic lineage cells of neoformed bone for the CTL and ALE groups in both periods (14 and 42 days). Alkaline phosphatase presented a lower staining area in the OST group compared to the CTL in both periods and the ALE at 42 days. Conclusion There was a decrease of osteocalcin precipitation at 42 days for the ALE and OST groups. Therefore, treatment with short-term sodium alendronate improved bone repair around the implants installed in the tibia of osteoporotic rats.

Kinetics of gene expression of alkaline phosphatase during healing of alveolar bone in rats.

Immunohistochemical studies and molecular biology have enabled us to identify numerous proteins that are involved in the metabolism of bone, and their encoding genes. Among these is alkaline phosphatase (ALP), an enzyme that is responsible for the initiation of mineralisation of the extracellular matrix during alveolar bone repair. To evaluate the gene expression of ALP during this process, we studied nine healthy adult male rats, which had their maxillary central incisors extracted from the right side and were randomly divided into three groups. During three experimental periods, 7 days, 14 days, and 28 days, the alveoli were curetted, the rats killed, and samples analysed by real-time reverse transcription polymerase chain reaction (qRT-PCR). The RNAm that encodes the gene for the synthesis of ALP was expressed during the three periods analysed, but its concentration was significantly increased at 14 and 28 days compared with at 7 days. There was no significant difference between 14 and 28 days (p=0.0005). We conclude that genes related to ALP are expressed throughout the healing process and more intensively during the later periods (14 and 28 days), which coincides with the increased formation of mineralised bone.