An interventional clinical trial investigating the effects of Spirulina platensis on dental fluorosis and antioxidant system in lambs reared in endemic areas

Abstract

This study aimed to evaluate the protective effect of Spirulina platensis primary against dental fluorosis and secondary against oxidative stress in lambs reared in endemic fluorosis areas. Forty-eight lambs aged 5 months were divided into four equal groups (each one including 6 males and 6 females). Groups I and II served as controls belonging respectively to fluorosis-free (Settat) and endemic fluorosis (El Fokra) areas, while the other two Groups III and IV (belonging to El Fokra) received respectively a fixed daily intake of 250 and 500 mg/kg bodyweight (BW) of Spirulina platensis. The experiment was carried out for 13 months until the adult incisors appeared for all animals. According to the Dean's Fluorosis Index (DFI), 500 mg/kg BW/day of Spirulina platensis (Group IV) protected against dental fluorosis. Moreover, in both male and female lambs, this dose significantly (p < 0.0001) reduced the plasmatic levels of fluoride, proteins, GSH, and MDA compared to the Group II. Furthermore, enzymatic activities of catalase and SOD increased significantly (p < 0.0001) in male and female lambs of the Group IV as compared to Group II. In conclusion, our findings support the potential use of Spirulina platensis as a valuable solution for addressing fluorosis in sheep, warranting further clinical trials.

Figure 1

Effect of Spirulina platensis on plasma fluoride level in lambs reared in endemic fluorosis area. Data were expressed as the mean ± SEM. Group I: lambs reared in fluorosis-free area, Group II: lambs reared in endemic fluorosis area, Group III: lambs supplemented with 250 mg/kg BW/day of Spirulina platensis, Group IV: lambs supplemented with 500 mg/kg BW/day of Spirulina platensis. A, B, C and D different superscripts within each column show significant differences at the level of plasma fluoride between the females of each group. a, b, c and d different superscripts within each column show significant differences at the level of plasma fluoride between the males of each group. *: indicates the significant difference at the level of plasma fluoride between males and females of each group.

Figure 2

Effect of Spirulina platensis on fecal fluoride levels in lambs reared in endemic fluorosis area. Data were expressed as the mean ± SEM. Group I: lambs reared in fluorosis-free area, Group II: lambs reared in endemic fluorosis area, Group III: lambs supplemented with 250 mg/kg BW/day of Spirulina platensis, Group IV: lambs supplemented with 500 mg/kg BW/day of Spirulina platensis. A: indicates that there is no significant difference in fecal fluoride levels between the females of each group. a indicates that there is no significant difference in fecal fluoride levels between the males of each group.

Figure 3

Effect of Spirulina platensis on plasma proteins level in lambs reared in endemic fluorosis area. Data were expressed as the mean ± SEM. Group I: lambs reared in fluorosis-free area, Group II: lambs reared in endemic fluorosis area, Group III: lambs supplemented with 250 mg/kg BW/day of Spirulina platensis, Group IV: lambs supplemented with 500 mg/kg BW/day of Spirulina platensis. A, B, C and D different superscripts within each column show significant differences at the level of plasma proteins between the females of each group. a, b, c and d different superscripts within each column show significant differences at the level of plasma proteins between the males of each group.

Figure 4

Effect of Spirulina platensis on reduced glutathione (GSH) level in lambs reared in endemic fluorosis area. Data were expressed as the mean ± SEM. Group I: lambs reared in fluorosis-free area, Group II: lambs reared in endemic fluorosis area, Group III: lambs supplemented with 250 mg/kg BW/day of Spirulina platensis, Group IV: lambs supplemented with 500 mg/kg BW/day of Spirulina platensis. A, B, C and D different superscripts within each column show significant differences at the level of GSH between the females of each group (p < 0.05). a, b, c and d different superscripts within each column show significant differences at the level of GSH between the males of each group.

Figure 5

Effect of Spirulina platensis on malondialdehyde (MDA) level in lambs reared in endemic fluorosis area. Data were expressed as the mean ± SEM. Group I: lambs reared in fluorosis-free area, Group II: lambs reared in endemic fluorosis area, Group III: lambs supplemented with 250 mg/kg BW/day of Spirulina platensis, Group IV: lambs supplemented with 500 mg/kg BW/day of Spirulina platensis. A, B, C and D different superscripts within each column show significant differences at the level of MDA between the females of each group. a, b, c and d different superscripts within each column show significant differences at the level of MDA between the males of each group.

Figure 6

Effect of Spirulina platensis on catalase (CAT) activity in lambs reared in endemic fluorosis area. Data were expressed as the mean ± SEM. Group I: lambs reared in fluorosis-free area, Group II: lambs reared in endemic fluorosis area, Group III: lambs supplemented with 250 mg/kg BW/day of Spirulina platensis, Group IV: lambs supplemented with 500 mg/kg BW/day of Spirulina platensis. A, B, C and D different superscripts within each column show significant differences at the level of CAT activity between the females of each group. a, b, c and d different superscripts within each column show significant differences at the level of CAT activity between the males of each group.

Figure 7

Effect of Spirulina platensis on superoxide dismutase (SOD) activity in lambs reared in endemic fluorosis area. Data were expressed as the mean ± SEM. Group I: lambs reared in fluorosis-free area, Group II: lambs reared in endemic fluorosis area, Group III: lambs supplemented with 250 mg/kg BW/day of Spirulina platensis, Group IV: lambs supplemented with 500 mg/kg BW/day of Spirulina platensis. A, B, C and D different superscripts within each column show significant differences at the level of SOD activity between the females of each group. a, b, c and d different superscripts within each column show significant differences at the level of SOD activity between the males of each group.

Figure 8

Effect of Spirulina platensis on hemoglobin (Hb) levels in lambs reared in endemic fluorosis area. Data were expressed as the mean ± SEM. Group I: lambs reared in fluorosis-free area, Group II: lambs reared in endemic fluorosis area, Group III: lambs supplemented with 250 mg/kg BW/day of Spirulina platensis, Group IV: lambs supplemented with 500 mg/kg BW/day of Spirulina platensis. A: indicates that there is no significant difference in hemoglobin levels between the females of each group. a indicates that there is no significant difference in hemoglobin levels between the males of each group.

Figure 9

Diagram presenting the experimental protocol.