Behavioural and cognitive effects of simvastatin dose used in stimulation of bone regeneration in rats

To analyze the effects of simvastatin (SVT) in the locomotion, anxiety and memory of rats, as a reflection of the administration of a minimum dose capable of stimulating bone regeneration in defects in the calvariae.

METHODS:

Surgical procedures were performed in 15 female Wistar rats, 2-month old, to insert the grafting material regenerator (Bone-ceramic^(r)) and/or SVT, followed by behavioural and cognitive assessments in the 7^th, 30^th and 60^th days post surgery.

RESULTS:

The SVT locally applied with the goal of bone regeneration in defects created in rat calvariae does not interfere with locomotion, anxiety levels and/or memories of rats, except for the first week following surgery, when an anxiolytic effect was observed, as a result of a possible central action.

CONCLUSION:

Failure to provoke any response within 30 and 60 days post surgical procedures suggests that SVT may constitute a good choice in stimulating bone regeneration without affecting the long term neural functions.

Cognition; Behavior; Simvastatin; Bone Regeneration; Rats

Authorship

Dircilei Nascimento de Sousa

Fellow Master degree, Postgraduate Program in Health Sciences, Health Sciences Faculty, Brasília University (UNB), Brasília-DF, Brazil. Surgical procedures, acquisition and interpretation of data, manuscript writingBrasília University BrazilBrasília, DF, BrazilFellow Master degree, Postgraduate Program in Health Sciences, Health Sciences Faculty, Brasília University (UNB), Brasília-DF, Brazil. Surgical procedures, acquisition and interpretation of data, manuscript writing

Washington Macedo de Santana

Fellow PhD degree, Postgraduate Program in Health Sciences, Health Sciences Faculty, UNB, Brasília-DF, Brazil. Surgical procedures, acquisition and interpretation of data, manuscript writingUNBBrazilBrasília, DF, BrazilFellow PhD degree, Postgraduate Program in Health Sciences, Health Sciences Faculty, UNB, Brasília-DF, Brazil. Surgical procedures, acquisition and interpretation of data, manuscript writing

Vania Moraes Ferreira

PhD, Full Professor, Health Sciences Faculty, UNB, Brasília-DF, Brazil. Statistical analysis, English language, critical revisionUNBBrazilBrasília, DF, BrazilPhD, Full Professor, Health Sciences Faculty, UNB, Brasília-DF, Brazil. Statistical analysis, English language, critical revision

Wagner Rodrigues Duarte

PhD, Full Professor, Health Sciences Faculty, UNB, Brasília-DF, Brazil. Conception and design of the study, manuscript writingUNBBrazilBrasília, DF, BrazilPhD, Full Professor, Health Sciences Faculty, UNB, Brasília-DF, Brazil. Conception and design of the study, manuscript writing

Correspondence: Dircilei Nascimento de Sousa Universidade de Brasília Faculdade de Ciências da Saúde Programa de Pós-Graduação de Ciências da Saúde Campus Universitário Darcy Ribeiro, s/n 70.910-900 Brasília-DF Brasil Tel.: (55 61)9984-0106 dircileinascimento@hotmail.com

Conflict of interest: none

Financial source: none

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Figures

Figures (5)

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FIGURE 1
Illustrations of the sequence of surgical procedures for the induction of bilateral defects in the calvariae of rats. A) Demarcation of the area where the bone defect is to be performed; B) Removal of the bone layer that characterizes the bone defect; C) Administration of a substance corresponding to what was chosen for each investigated group, D) area procedure finalized and ready to be sutured.

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FIGURE 2
Schematic representation of the steps of the experimental procedures adopted in the research. A) The surgeries took place on day 1 of the beginning of the experiments, and behavioural evaluations occurred on the 7th, 30th and 60th days after anaesthesia and surgery procedures in order to verify the level of stress/anxiety and some cognition impairment; B) The tests used in the research were open field (locomotion); elevated plus-maze (anxiety); and "step down" inhibitory avoidance (memory) tests.

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FIGURE 3
Effects of simvastatin (0.1 mg/kg) on locomotor activity of rats evaluated in the open field test for 5 min, on the 7th, 30th and 60th days post surgery. The results are expressed as mean ± SEM of five animals per group. G1= control group; G2= left side: BoneCeramic(r) (BC)/right side: BoneCeramic(r) + simvastatin (BC+SVT); G3= left side: Collagen sponge (CS)/right side: Collagen sponge + simvastatin (CS+SVT).

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FIGURE 4
Effects of simvastatin (0.1 mg/kg) on the anxiety-like behaviour of rats evaluated in the elevated plus maze test on the 7th, 30th and 60th days post surgery. The results are expressed as mean ± SEM of five animals per group. *p≤0.05 compared to the control group (G1) on the 7th day post surgery (One way ANOVA; Newman-Keuls test). G1= control group; G2= left side: BoneCeramic(r) (BC)/ right side: BoneCeramic(r) + simvastatin (BC+SVT); G3= left side: Collagen sponge (CS)/ right side: Collagen sponge + simvastatin (CS+SVT).

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FIGURE 5
Effects of simvastatin (0.1 mg/kg) on short- and long-term memories of rats evaluated in the "step-down" inhibitory avoidance test on the 7th, 30th and 60th days post surgery. The results are expressed as mean ± SEM of 5 animals per group. STM was evaluated 1h after the training. LTM was evaluated 24h after STM on the 7th day post surgery, and again on the 30th and 60th days. *p≤0.05 compared to the control group (G1) on 7th day post surgery (Kruskal-Wallis test, followed by Dunn's multiple comparison). G1= control group; G2= left side: BoneCeramic(r) (BC)/ right side: BoneCeramic(r) + simvastatin (BC+SVT); G3= left side: Collagen sponge (CS)/ right side: Collagen sponge + simvastatin (CS+SVT).

FIGURE 1 Illustrations of the sequence of surgical procedures for the induction of bilateral defects in the calvariae of rats. A) Demarcation of the area where the bone defect is to be performed; B) Removal of the bone layer that characterizes the bone defect; C) Administration of a substance corresponding to what was chosen for each investigated group, D) area procedure finalized and ready to be sutured.

FIGURE 2 Schematic representation of the steps of the experimental procedures adopted in the research. A) The surgeries took place on day 1 of the beginning of the experiments, and behavioural evaluations occurred on the 7th, 30th and 60th days after anaesthesia and surgery procedures in order to verify the level of stress/anxiety and some cognition impairment; B) The tests used in the research were open field (locomotion); elevated plus-maze (anxiety); and "step down" inhibitory avoidance (memory) tests.

FIGURE 3 Effects of simvastatin (0.1 mg/kg) on locomotor activity of rats evaluated in the open field test for 5 min, on the 7th, 30th and 60th days post surgery. The results are expressed as mean ± SEM of five animals per group. G1= control group; G2= left side: BoneCeramic(r) (BC)/right side: BoneCeramic(r) + simvastatin (BC+SVT); G3= left side: Collagen sponge (CS)/right side: Collagen sponge + simvastatin (CS+SVT).

FIGURE 4 Effects of simvastatin (0.1 mg/kg) on the anxiety-like behaviour of rats evaluated in the elevated plus maze test on the 7th, 30th and 60th days post surgery. The results are expressed as mean ± SEM of five animals per group. *p≤0.05 compared to the control group (G1) on the 7th day post surgery (One way ANOVA; Newman-Keuls test). G1= control group; G2= left side: BoneCeramic(r) (BC)/ right side: BoneCeramic(r) + simvastatin (BC+SVT); G3= left side: Collagen sponge (CS)/ right side: Collagen sponge + simvastatin (CS+SVT).

FIGURE 5 Effects of simvastatin (0.1 mg/kg) on short- and long-term memories of rats evaluated in the "step-down" inhibitory avoidance test on the 7th, 30th and 60th days post surgery. The results are expressed as mean ± SEM of 5 animals per group. STM was evaluated 1h after the training. LTM was evaluated 24h after STM on the 7th day post surgery, and again on the 30th and 60th days. *p≤0.05 compared to the control group (G1) on 7th day post surgery (Kruskal-Wallis test, followed by Dunn's multiple comparison). G1= control group; G2= left side: BoneCeramic(r) (BC)/ right side: BoneCeramic(r) + simvastatin (BC+SVT); G3= left side: Collagen sponge (CS)/ right side: Collagen sponge + simvastatin (CS+SVT).

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Behavioural and cognitive effects of simvastatin dose used in stimulation of bone regeneration in rats

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[1] Correspondence: Dircilei Nascimento de Sousa Universidade de Brasília Faculdade de Ciências da Saúde Programa de Pós-Graduação de Ciências da Saúde Campus Universitário Darcy Ribeiro, s/n 70.910-900 Brasília-DF Brasil Tel.: (55 61)9984-0106 dircileinascimento@hotmail.com