Next Article in Journal
Protective Immunity Induced by an Eimeria tenella Whole Sporozoite Vaccine Elicits Specific B-Cell Antigens
Next Article in Special Issue
Growth Rate and Bone Hydroxyproline Concentration in Turkeys Fed with a Silage-Composed Diet Modified with Different Diet Cation–Anion Differences (DCADs)
Previous Article in Journal
Case Study: An Evaluation of Detection Dog Generalization to a Large Quantity of an Unknown Explosive in the Field
Previous Article in Special Issue
Relationship between Bone Quality, Egg Production and Eggshell Quality in Laying Hens at the End of an Extended Production Cycle (105 Weeks)
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Animals
Volume 11
Issue 5
10.3390/ani11051343
animals-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Camelina Oil Supplementation Improves Bone Parameters in Ovariectomized Rats

by
Iwona Puzio
1,*,
Dorota Graboś
1,
Marek Bieńko
1,
Radosław P. Radzki
1,
Aneta Nowakiewicz
2 and
Urszula Kosior-Korzecka
2
1
Department of Animal Physiology, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, Akademicka St. 12, 20-950 Lublin, Poland
2
Department of Preclinical Veterinary Sciences, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, Akademicka St. 12, 20-950 Lublin, Poland
*
Author to whom correspondence should be addressed.
Animals 2021, 11(5), 1343; https://doi.org/10.3390/ani11051343
Submission received: 30 March 2021 / Revised: 28 April 2021 / Accepted: 3 May 2021 / Published: 9 May 2021
(This article belongs to the Collection Gut and Bone in Health and Disease)
Versions Notes

Abstract

:

Simple Summary

Supplementation of ovariectomized rats with Camelina sativa oil, which is rich in polyunsaturated fatty acids (PUFA), especially n-3 family fatty acids, can be an effective way to improve bone parameters. Administration of 5 g/kg body weight and 9 g/kg body weight of camelina oil to rats suppressed a decrease in densitometric, tomographic, and strength parameters of femurs and an increase in the serum level of the C-terminal telopeptide of type I collagen caused by estrogen deficiency. Furthermore, ovariectomized rats receiving camelina oil were characterized by a high level of osteocalcin.

Abstract

The aim of the present study was to determine the effect of administration of Camelina sativa oil (CO) as a source of polyunsaturated fatty acids (PUFA) on bone parameters in ovariectomized rats (OVX). Overall, 40 10-week-old healthy female Wistar rats were divided into 4 groups with 10 animals in each. Rats in the control group (SHO) were subjected to a sham operation, whereas experimental rats (OVX) were ovariectomized. After a 7-day recovery period, the SHO the rats received orally 1 mL of physiological saline for the next 6 weeks. The OVX rats received orally 1 mL of physiological saline (OVX-PhS), 5 g/kg BW (OVX-CO5), or 9 g/kg BW (OVX-CO9) of camelina oil. The use of camelina oil had a significant effect on body weight, lean mass, and fat mass. The camelina oil administration suppressed the decrease in the values of some densitometric, tomographic, and mechanical parameters of femur caused by estrogen deficiency. The CO treatment increased significantly the serum level of osteocalcin and decreased the serum level of C-terminal telopeptide of type I collagen in the OVX rats. In conclusion, camelina oil exerts a positive osteotropic effect by inhibiting ovariectomy-induced adverse changes in bones. Camelina oil supplementation can be used as an efficient method for improving bone health in a disturbed state. However, further research must be carried out on other animal species supplemented with the oil.

1. Introduction

Pathological bone and cartilage changes are an increasing problem in both livestock and companion animals. The health conditions include osteopenia with different etiologies, osteoporosis, osteochondrosis, rickets, and osteoarthritis [1,2,3,4]. In both groups of animals, they cause movement disorders and pain and reduce the quality of their life. In the case of pigs and poultry, they lead to production losses.
Hence, to improve the health of bones and minimize production losses, intensive studies are conducted to identify factors that can act preventively or therapeutically in bone diseases. One of the possibilities is the use of botanical compounds. In recent years, oilseeds and plant oils have aroused great interest. They are a source of n-3 and n-6 polyunsaturated fatty acids (PUFA), which have received considerable attention for their role in mitigation of bone disorders. Most PUFAs from the n-3 and n-6 families are not synthesized by mammals. Therefore, their precursors, i.e., such essential acids as α-linolenic acid (ALA, 18:3 n-3) and linoleic acid (LA, 18:2 n-6), must be supplied with food. The involvement of elongases and desaturases leads to formation of eicosapentaenoic acid (EPA, 20:5 n-3), docosahexaenoic acid (DHA, 22:6 n-3), and arachidonic acid (AA, 20:4 n-6). EPA and AA are substrates for production of eicosanoids; they compete for the same enzymes in the metabolic pathways [5,6]. Eicosanoids derived from n-3 PUFAs (3-series prostaglandins, 3-series thromboxanes, 5-series leukotrienes, E-series resolvins) generally have anti-inflammatory effects, whereas those produced from n-6 PUFAs (2-series prostaglandins, 2-series thromboxanes, 4-series leukotrienes) act as pro-inflammatory agents [7]. Eicosanoids derived from n-6 PUFAs influence the synthesis of pro-inflammatory cytokines, i.e., interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), which participate in the development of such skeletal disorders as osteopenia, osteoporosis, or arthritis [8].
There are many papers in the databases on the impact of various oils of plant and animal origin, as well as individual PUFAs on bone metabolism and properties in both animals and humans. A positive effect of seeds, flax oil, soybean oil, sunflower oil, sesame oil, and olive oil on bone tissue has been demonstrated [9,10,11,12,13,14]. However, there are also studies indicating a controversial impact of these oils [12,15,16,17].
In recent years, bioproducts from Camelina sativa, i.e., seeds, oil, and cake, have aroused considerable interest. Camelina sativa, a species from the Brassicaceae family also called false flax, gold-of-pleasure, wild flax, linseed dodder, German sesame, or Siberian oilseed, is one of the oldest cultivated plants. The history of cultivation of camelina goes back to the Bronze Age when this plant was used for consumption in Europe [18]. Camelina is a very fast growing annual or winter annual plant. The advantages of camelina include its relatively low habitat and agrotechnical requirements [19]. It grows in ruderal habitats and in arable fields. This plant resembles a weed: it has very low soil requirements and high drought tolerance. Camelina, therefore, offers lower production costs than other oilseed plants [20]. The increasing interest in the false flax in recent years is related to the high content of unsaturated fatty acids (around 90%) and PUFAs (50–60%), which are very valuable for animal and human health [21,22]. It has been found that the fatty acid composition of camelina oil is more favorable than the composition of a number of other plant oils, such as olive, flax, rapeseed, and sunflower oils. Camelina oil is characterized by high content of ALA (35–50%), lower content of LA (15–24%), and high content of tocopherols (about 800 mg/kg) and phenolic compounds, which results in greater oxidative stability compared to other oils containing large amounts of unsaturated acids [21,22,23,24,25,26,27]. Regardless of the fatty acids, the high nutritional value of camelina oils is also related to the content of other bioactive compounds such as tocopherols, carotenoids, and phytosterols, which lower the cholesterol level in blood, as well as optimal nutritional quality indices: the thrombogenicity index and the atherogenicity index [24,26,27]. Camelina oil also contains anti-nutritional substances like erucic acid and glucosinolates [21,22]. In an animal study, camelina bioproducts were found to influence positively the PUFAs content in eggs and meat of broiler chickens and rabbits, which is beneficial for human nutrition [28,29,30]. In quail, camelina was reported to prevent lipid oxidation without any adverse effect on performance and carcass characteristics [31]. Moreover, camelina oil in combination with conjugated linoleic acid improved bone densitometric parameters in broiler chickens [32]. However, there is no research on its effect on bone tissue when it is supplemented alone in mammals. Camelina oil also increased plasma n-3 fatty acid content and reduced plasma n-6 fatty acid content and serum triglyceride levels in pigs [33]. However, higher content of camelina bioproducts in food was found to cause a decrease in weight and feed intake, as well as poor feed conversion in animals [34,35]. With the properties mentioned above, camelina has been increasingly being appreciated for the last 20 years.
However, the influence of camelina bioproducts as a source of PUFAs on the metabolism of the skeletal system is very poorly known. Considering the prevalence of bone and joint disorders in animals and the composition of PUFAs in camelina oil, it can be assumed that supplementation with this product may positively influence bone tissue. Thus, the present study aimed to identify the potential osteotropic effects of camelina oil in female rats with experimental osteopenia induced by bilateral ovariectomy.

2. Materials and Methods

2.1. Animals and Experimental Design

In total, 40 10-wk-old healthy female Wistar rats (initial BW of approximately 220–240 g) were used in the study. Before the experiment, the rats were acclimatized to the new environmental conditions of the vivarium for 7 days. During the experiment, the rats were housed in a room with controlled temperature (22 ± 2 °C) and humidity (55 ± 10%) under a 12:12 h light–dark cycle with access to commercial diet for laboratory animals (Agropol, Motycz, Poland) and water at all times (except for a period of overnight fasting prior to the surgery and euthanasia). The rats were kept in plastic cages with sawdust bedding. On the day of the surgery, the rats were anaesthetized with an intramuscular injection of ketamine (Biowet-Puławy, Puławy, Poland), xylazine (SPOFA, Prague, Czech Republic), and atropinum sulphuricum (Polfa-Warszawa, Warsaw, Poland) at the doses of 10, 10, and 0.1 mg/kg of BW, respectively, and submitted to a sham operation (SHO; n = 10) or an ovariectomy (OVX) (n = 30). During the SHO operation, an incision was made in the abdominal wall, the viscera were repositioned manually, and the wound was sutured. In turn, the ovaries were removed during OVX. After the surgery, the rats were placed individually in plastic cages for a 7-day recovery period. No symptoms of postoperative complications were observed. No analgesics were administered to the rats during the postoperative period. Subsequently, the OVX rats were randomly divided into three groups receiving orally 1 mL of physiological saline (OVX-PhS) (n = 10) or camelina oil in the amount of 5 g/kg/BW (OVX-CO5; n = 10) or 9 g/kg/BW (OVX-CO9; n = 10) once a day for 6 weeks of the experiment. The doses were selected based on the literature data on the use flax oil, which is also characterized by a high ALA content. To determine the amount of oil administered, the BW of the rats was monitored every two days. The control rats (SHO; n = 10) received orally 1 mL of physiological saline. Animal plastic feeding tubes (Fuchigami, Kyoto, Japan) were used for the oral administration of oil or PhS. After 6 weeks of the experimental treatment, overnight fasted rats were weighted and euthanized by CO2 overdose. Then, blood was collected by cardiac puncture, and euthanasia was confirmed by cervical dislocation. Blood samples were collected into sterile tubes for clotting and then centrifuged at 3000 rpm for 30 min. Some serum samples were kept at −70 °C and others were stored at −20 °C until analysis. After euthanasia, the body composition, bone mineral density, and bone mineral content of total skeleton were determined. Afterwards, the right femora were dissected, cleaned of soft tissues, weighed, and frozen at −20 °C until further analysis.

2.2. Analysis of Fatty Acids in Camelina Oil

The fatty acids in the camelina oil were analyzed as methyl esters by gas chromatography. Fatty acid methyl esters (FAME) were prepared according to regulation No. PN-ISO 5509:1996 [36]. FAME analysis was performed using a gas chromatograph Agilent 6890N (Agilent Technologies, Santa Clara, CA, USA) equipped with a flame ionization detector and a HP-SSAP capillary column (30 m × 0.53 mm × 1 µm) according to PNEN ISO 5508:1996 [37]. The results for each fatty acid were expressed as % of total FAME. The composition of fatty acids in the camelina oil is presented in Table 1.

2.3. Densitometric Analysis (DXA) of the Body Composition, Whole Skeleton, and Isolated Femur

Bone mineral density (Tot.BMD) and bone mineral content (Tot.BMC) of the total skeleton and isolated right femora (fBMD, fBMC) were determined with the dual-energy X-ray absorptiometry method (DXA) using a Norland Excell Plus Densitometer (Norland, Fort Atkinson, WI, USA) equipped with Norland Illuminatus Small Subject Scan software v. 4.3.1. dedicated to small animals. The DXA method was also used for evaluation of the body composition parameters, e.g., lean mass (LM) and fat mass (FM). LM, FM, Tot.BMD, and Tot.BMC were measured immediately after euthanasia. Previously frozen femurs were kept at room temperature for 12 h before scanning. The scanning was performed using the following parameters: scout scan speed 100 mm/s, resolution 3.0 × 3.0 mm; measurement scan speed 10 mm/s, resolution 1.0 × 1.0 mm [38]. The region of interest (ROI) after the scout scan was defined manually. The densitometer was calibrated before every measurement series using quality assurance phantoms provided by the manufacturer in agreement with established procedures. To ensure credibility, all scans were performed by the same operator.

2.4. Peripheral Quantitative Computed Tomography (pQCT) of Isolated Femur

The right femurs were scanned by peripheral quantitative computed tomography (pQCT) using an XCT Research SA Plus system with software version 6.2.C (Stratec Medizintechnik GmbH, Pforzheim, Germany) [38]. The scans were performed in the middle diaphysis (at 50% of bone length) for the analysis of shaft microarchitecture and cortical bone tissue. Total bone mineral content (T.BMC), total volumetric bone mineral density (T.vBMD), total bone area (T.Ar), cortical bone area (Ct.Ar), cortical bone mineral content (Ct.BMC), cortical volumetric bone mineral density (Ct.vBMD), cortical thickness (Ct.Th), periosteal (Peri.C), and endocortical (Endo.C) circumferences, polar moment of inertia of cortical bone (PMI), and strength-strain index (SSI) were determined. Femur diaphysis was tested with the threshold set at 0.790 cm−1 and at cortical mode 2. Initial scan was performed at a speed of 10 mm/s, while proper scan was carried out at a speed of 4 mm/s. The pQCT system was calibrated before every measurement series with the use of the quality assurance phantom (pQCT QA-Phantom).

2.5. Mechanical Testing

After DXA and pQCT scanning, the mechanical parameters of isolated femurs were determined by a 3-point bending test with the use of a Zwick–Roell Testing Machine Z010 (Zwick–Roell GmbH and Co. KG, Ulm, Germany) as described earlier [38]. The apparatus was equipped with a measuring head (XForce HP series) with an operation range from 0 to 1 kN at a constant speed of 10 mm/min. After measuring the length, the bones were placed on two supports, and the measuring head loaded the midshaft perpendicularly to the horizontal axis at 50% of bone length. The distance between the supports was set at 40% of femur length. The data were analyzed with testXpert II 3.1 software (Zwick–Roell GmbH and Co. KG, Ulm, Germany). The ultimate force, work to fracture, and Young’s modulus were determined.

2.6. Markers of Bone Metabolism

Serum interleukin-6 (IL-6), osteocalcin, and C-terminal telopeptide of type I collagen (CTX-I) concentrations were measured using a commercial Rat IL-6 ELISA kit (DIACLONE SAS, Besançon, France, No. 670.010.096), a Rat MIDTM Osteocalcin EIA kit (Immunodiagnostic Systems Ltd., Boldon, Tyne and Wear, UK, No. AC-12F1), and a RatLaps CTX-ITM EIA kit (Immunodiagnostic Systems Ltd., Boldon, Tyne and Wear, UK, No. AC-06F1), respectively. The analysis of bone metabolism markers was performed using a Benchmark Plus microplate reader equipped with MicroplateManager software Version 5.2.1 (Bio-Rad Laboratories Inc., Hercules, CA, USA).

2.7. Statistical Aanalysis

All data were presented as mean values ± S.E.M. A one-way analysis of variance (ANOVA) and Tukey’s test were used to test the significant differences among the groups. For all comparisons, p < 0.05 was considered as statistically significant. The analysis of significant differences was performed with the use of STATISTICA 13.1 software (StatSoft, Inc. Tulsa, OK, USA).

3. Results

3.1. Body Weight, Body Composition, Bone Mass, and Length

The final BW and LM were significantly higher in the experimental rats than in the SHO group (Table 2). The BW in the CO-supplemented groups was by 10% higher than in the SHO group. There were no significant differences in BW and LM between the OVX-PhS and OVX rats treated with CO. The largest enhancement of LM was observed in the OVX-CO5 group (by 24% vs. SHO). Moreover, all experimental groups were characterized by higher values of FM, compared to the SHO animals. However, significant differences in comparison to the control animals were found in the OVX-CO9 group. The CO treatment significantly increased femur weight, compared to the SHO and OVX- PhS rats (Table 2).

3.2. DXA Parameters of Whole Skeleton and Femur

The values of the DXA parameters of the whole skeleton and isolated femurs are presented in Table 2. The ovariectomy significantly decreased total skeleton BMC and BMD by 12% and 8%, respectively, in comparison to these values in the SHO rats. Both groups receiving CO were characterized by higher Tot.BMC than the OVX-PhS group, i.e., by 12% and 18%, respectively. The values of this parameter were similar to those in the SHO group. Statistically significant differences in the Tot.BMD values were observed between the OVX-PhS and SHO groups. No significant differences in Tot.BMD were found between the SHO and CO-treated groups. The values of femur BMC and BMD were significantly lower in the OVX-PhS group than in the SHO and both OVX-CO groups, i.e., by 10%, 12%, and 11% for fBMC and by 3.5%, 7.2%, and 5.6% for fBMD, respectively. The values of fBMD and fBMC in the SHO and OVX-CO groups were similar.

3.3. pQCTMeasurements of Femur

The results of the pQCT measurement of cortical bone tissue in the femur midshaft are shown in Table 3. The lowest values of most of the analyzed parameters were observed in the OVX-PhS group. The camelina oil treatment significantly elevated T.vBMD, T.BMC, Ct.A, and Ct.Th, compared to these parameters in the OVX-PhS rats. The bone parameters were increased in the OVX-CO5 and OVX-CO9 groups as follows: T.vBMD—by 11.5% and 8%; T.BMC—by 8% and 9%; Ct.Ar—by 8% and 7%; and Ct.Th—by 10% and 8%, respectively. Moreover, Ct.BMC in the OVX-CO9 group was by 9% higher in comparison to the OVX-PhS group.

3.4. Mechanical Parameters of Femur

The results of the mechanical tests of bone are presented in Table 2. Although there were no statistically significant differences between the OVX-PhS and SHO rats, the values of the tested parameters were lower in the OVX rats in all cases. The ultimate force was significantly increased in the OVX-CO5 and OVX-CO9 groups in comparison to the OVX-PhS group. The values of ultimate force in the OVX-CO5 and OVX-CO9 groups were by 84% and 64% higher than in the OVX-PhS group. In turn, the values of work to fracture were also significantly higher in the OVX animals treated with CO than in those receiving PhS. Neither the ovariectomy nor the camelina oil treatment affected the Young’s modulus significantly.

3.5. Biochemical Markers of Bone Metabolism

The values of the biochemical markers of bone metabolism are presented in Table 4. The lowest serum osteocalcin concentration was noted in the control SHO rats. The ovariectomy caused an increase in the osteocalcin concentration. A significantly higher concentration of osteocalcin was observed in the CO-treated ovariectomized rats than in the SHO and OVX-PhS rats. The serum CTX-I concentration was significantly elevated in the OVX-PhS animals in comparison to the SHO and CO-treated groups. The serum concentration of CTX-I was by 63%, 36%, and 36% higher in OVX-PhS than in SHO, OVX-CO5, and OVX-CO9, respectively. The IL-6 concentration increased in the OVX-PhS animals compared with the SHO rats. No significant changes were observed between the CO-treated groups and the other groups. However, the concentration of IL-6 was lower in both CO groups than in the OVX-PhS animals.

4. Discussion

Ovariectomy results in deficient estrogen production and consequent negative changes in bone metabolism, leading to bone loss and increased susceptibility to fractures. However, a number of mechanisms may contribute of this effect. Ovary removal in rats also results in progressive bone loss and changes in bone parameters. Such changes in the densitometric, tomographic, and strength parameters were earlier found in experimental animals [39,40,41].
In the present study, the OVX rats exhibited significant bone loss manifested in the reduction in Tot.BMD, Tot.BMC, fBMD, fBMC, work to fracture, and T.vBMD. Moreover, the changes in the Il-6 and CTX-I levels detected in this study confirm the negative effect of ovariectomy on bone metabolism. In the OVX rats, the IL-6 and CTX-I levels were elevated in comparison to those in the control rats. Ovariectomy stimulates the IL-6 production, which is known to be up-regulated in a state of estrogen deficiency [42], and leads to an increase in the CTX-I concentration. Estrogen deficiency can also increase the concentration of IL-1, TNF-α [42], and other bone metabolism markers, i.e., bone alkaline phosphatase (bALP) as a marker of bone formation and bone disorders, as well as tartrate-resistant acid phosphatase (TRAP) as a bone resorption marker [43].
The results of our study indicate that the camelina oil treatment reduced the ovariectomy-stimulated negative changes in bone tissue. After the application of CO, we observed suppression in osteoclast-pathway activity, as evidenced by the suppression of the decrease in the values of Tot.BMC, T.vBMD, fBMD, fBMC, and work to fracture caused by estrogen deficiency. Moreover, the CO treatment restored the elevated level of the CTx values in the OVX rats to the SHO values. In a human study, a significant decrease in the CTX level was found after supplementation with n-3 PUFAs [44]. The decrease in bone resorption markers was also observed by Griel et al. [45]. In a study conducted by Boulbaroud et al. [43], flaxseed and sesame oil supplementation decreased bALP and TRAP levels. However, there are also literature data that indicate no changes in bone resorption and formation markers in healthy individuals [46].
Based on the present results, it can be assumed that CO also affects bone formation. This is evidenced by the higher values of some parameters in the OVX oil-receiving animals compared to the SHO group. Primarily, the OVX rats receiving CO had significantly higher serum osteocalcin levels than the SHO animals. This increase in the osteocalcin concentration may evidence bone turnover on the one hand and bone formation on the other. However, the simultaneous increase in the values of bone mass, Ct.Th, Ct.BMC, and T.vBMD indicates stimulation of bone formation.
Previous studies have shown that n-3 PUFAs supplementation reduced bone loss in OVX rats [47,48]. Moreover, a positive effect of dietary soybean, as well as flaxseed and sesame oils, on femur bone density and calcium content in bone ash was observed in osteoporotic rats by Wahba et al. [9]. In turn, a positive effect of flaxseed and sesame oils connected with partial mitigation of osteoporotic changes in OVX rats was described by Boulbaroud et al. [43]. A beneficial impact of flaxseed oil and soybean oil on the prevention of negative changes in bone rats was also reported by Hassan et al. [10] and El-Saeed et al. [13]. As shown by Elbahnasawy et al. [14], enhanced bone formation and reduction in bone loss markers, as reflected by reduced CTX levels in plasma, was observed after administration of soybean and flaxseed oils in a GC-induced osteoporosis model. Thus, our results are in line with other studies.
There are several mechanisms by which n-3 and n-6 fatty acids affect bone tissue. For instance, PUFAs can modulate calcium absorption, production of inflammatory cytokine and prostaglandins, and regulation of osteoblast and osteoclast activity by changing the fatty acid composition in bone cell membranes [49,50]. n-3 PUFAs have been reported to suppress osteoclast activity and improve osteoblast activity [51,52]. PUFAs influence intestinal calcium absorption via both active and passive transport. DHA, but not EPA and AA, stimulated active calcium transport by increasing Ca2+-ATPase activity in the basolateral membrane of duodenal enterocytes [53]. However, supplementation with fish oil and evening primrose oil also resulted in an increase in calcium transport across the basolateral membrane [54]. The changes in calcium transport are caused by the incorporation of long-chain PUFAs into the cell membrane. On the one hand, this leads to increased membrane fluidity, permeability, and speed of flip-flop transport across the membrane and, in consequence, cellular uptake of different molecules, particularly via passive transport [55,56]. On the other hand, increased membrane unsaturation alters the dispersion of lipid rafts within membranes and can modulate the activity of membrane proteins, e.g., Ca2+-ATPase [57,58].
Inflammatory cytokines play a crucial role in the pathogenesis of osteoporosis by stimulation of bone resorption and inhibition of osteosynthesis. IL-1, IL-6, and TNF-α are involved in bone metabolism [8]. n-6 PUFAs increase the synthesis of proinflammatory cytokines by bone cells, whereas n-3 acids attenuate this effect [45,59]. Upon increasing dietary intake of n-3 PUFAs, n-3 acids partially replace n-6 acids in the cell membranes, and the increasing n-3/n-6 PUFA ratio may result in changes in membrane function, and reduction in the synthesis of proinflammatory factors [60,61,62]. Flaxseed oil supplementation was reported to reduce the TNF-α level [45,63]. In turn, no changes in IL-1 and IL-6 levels after flaxseed and walnut oil supplementation were observed [45]. In our study, we investigated the serum level of IL-6, which was elevated by the ovariectomy. The camelina oil supplementation reversed this effect. However, these results were not statistically significant. Il-6 is known to promote the development of osteoclasts and to stimulate bone resorption [8]. Its production was stimulated by 2-series prostaglandins (PGE2) [64,65] and Il-1 [66]. PGE2 is one of the modulators of bone remodeling. It has been shown to mediate both bone formation and resorption [67,68]. At a physiological concentration, PGE2 positively influences bone growth [69,70].
Modification of dietary PUFAs leads to alterations in prostaglandin metabolism [71,72,73]. PUFAs n-6 and n-3 are precursors for PGE2 and PGE3, respectively. Dietary PUFAs, particularly the n-6/n-3 ratio, are important factors for the bone marrow fatty acid profile, and this in turn determines the capacity of bone for synthesis of PGE2 [72,73,74]. It was found that a rise in the dietary n-6/n-3 ratio was accompanied by an increase in the level of PGE2 and a decline in Ca, P, and Mg contents in bone [73]. On the other hand, a low ratio of n-6/n-3 PUFAs decreased PGE2 production and improved bone formation [69]. AA, cleaved from membrane phospholipids of osteoblasts, is a substrate of cyclooxygenase-2 (COX-2) for PGE2 synthesis. Some data indicate that n-6 PUFAs up-regulate COX-2 expression and stimulate production of PGE2, whereas n-3 PUFAs diminish COX-2 expression [75]. An increase in the n-3 PUFAs content in diet may lead to displacement of AA from phospholipids and reduction in PGE2 synthesis [71,76]. PGE3 synthesized from EPA also mediates bone cell function. However, AA is more easily synthesized to PGE2 than EPA is to PGE3 [77]. The reduction in PGE2 was observed in rats after flaxseed and menhaden oil administration, but it was not accompanied by changes in femur area, BMC, and BMD [72]. Similar results were observed in chicken receiving flaxseed oil alone or in combination with palm oil [78]. The decreased PGE2 concentration was not associated with changes in bone characteristics. However, flaxseed oil supplementation led to an increase in the ALP concentration and a decrease in the TRAP concentration.
A high level of PGE2 leads to bone resorption [79,80]. This may be related to stimulation of the expression of the receptor activator of nuclear factor NF-kB ligand (RANKL) on osteoblast and its receptor (receptor activator of nuclear factor NF-kB, RANK) on osteoclasts and reduction in the expression of the decoy receptor, osteoprotegerin (OPG), on osteoblasts by the high level of PGE2 [81,82]. The binding of RANKL to OPG prevents osteoclastogenesis, whereas binding to RANK stimulates this process [83]. Currently, the balance of RANKL/OPG is considered very important for bone metabolism and the pathogenesis of resorptive bone disease [84]. Thus, a high level of PGE2 has a stimulating effect on osteoclastogenesis and OC activity [85,86]. It has been found that AA leads to enhanced synthesis of RANKL, which binds to RANK on osteoclast precursors stimulating osteoclastogenesis and maturation of osteoclasts [79]. Moreover, AA inhibits the synthesis of OPG [82]. PGE3 synthesized from EPA also promotes osteoclastogenesis but the conversion EPA to PGE3, as mentioned before, is less efficient.
The favorable effect of n-3 acids on bone tissue was observed not only in rats, but also in other animal species [87]. Higher values of the cortical wall thickness, cross-sectional area, cortical index, BMC, BMD, maximum elastic strength, and maximum strength were determined for femurs of pigs after linseed oil supplementation [88]. Flax oil alone or in combination with palm oil had a positive effect on the biomarkers of bone growth and increased tibia calcium levels in broiler chickens [78]. However, no significant differences in BMD, BMC, strength parameters, and ash percentage after flaxseed supplementation were observed in piglets and chickens [15,89,90]. The source of PUFAs, i.e., the type of oil, seems to be important for the impact of PUFAs on bone tissue. Equally important is the ratio of n-3 to n-6 PUFAs. Nevertheless, the literature data on the effect of individual vegetable oils on animal bones are not explicit.

5. Conclusions

Our research has shown that camelina oil has a positive osteotropic effect via inhibition of adverse changes in bones caused by ovariectomy. The administration of the oil yielded similar or even better densitometric, tomographic, and mechanical parameters of the bones in the OVX rats in comparison with the values in the control rats. These results are confirmed by the changes in the concentration of bone metabolism markers. The decrease in the serum concentration of CTX-I with the simultaneous increase in the concentration of osteocalcin indicates suppression of bone tissue resorption by inhibition of the osteoclastic-pathway on the one hand and an increase in bone turnover with a simultaneous advantage of bone formation over resorption on the other hand. Camelina oil supplementation can be an efficient method for improving bone health in a disturbed health state. However, further research must be carried out on other animal species supplemented with this oil. In addition, it is also necessary to investigate the effect of camelina oil on bones in individuals without bone changes.

Author Contributions

Conceptualization, I.P. and D.G.; methodology, I.P. and D.G.; investigation, I.P. and D.G.; formal analysis, I.P., D.G., M.B., R.P.R., A.N. and U.K.-K.; project administration, I.P. and D.G.; supervision, I.P.; writing—original draft preparation, I.P., A.N. and U.K.-K. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the 2nd Local Animal Welfare Committee at the University of Life Sciences in Lublin, Poland (82/2009). All procedures complied with the Guiding Principles for Research Involving Animals.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Jensen, T.B.; Toft, N. Causes of and predisposing risk factors for leg disorders in growing-finishing pigs. CAB Rev. Perspect. Agric. Vet. Sci. Nutr. Nat. Res. 2009, 4, 1–8. [Google Scholar] [CrossRef]
  2. Madson, D.M.; Ensley, S.M.; Gauger, P.C.; Schwartz, K.J.; Stevenson, G.W.; Cooper, V.L.; Janke, B.H.; Burrough, E.R.; Goff, J.P.; Horst, R.L. Rickets: Case series and diagnostic review of hypovitaminosis D in swine. J. Vet. Diag. Investig. 2012, 24, 1137–1144. [Google Scholar] [CrossRef] [Green Version]
  3. Mohsina, A.; Zama, M.M.S.; Tamilmahan, P.; Gugjoo, M.B.; Singh, K.; Gopinathan, A.; Gopi, M.; Karthik, K. A retrospective study on incidence of lameness in domestic animals. Vet. World 2014, 7, 601–604. [Google Scholar] [CrossRef] [Green Version]
  4. Akyuz, H.C.; Onbasilar, E.E. Non-infectious skeletal disorders in broilers. Worlds Poult. Sci. J. 2020, 76, 611–623. [Google Scholar] [CrossRef]
  5. Ratnayake, W.M.; Galli, C. Fat and fatty acid terminology, methods of analysis and fat digestion and metabolism: A background review paper. Ann. Nutr. Metab. 2009, 55, 8–43. [Google Scholar] [CrossRef]
  6. Turchini, G.M.; Francis, D.S. Fatty acid metabolism (desaturation, elongation and b-oxidation) in rainbow trout fed fish oil- or linseed oil-based diets. Br. J. Nutr. 2009, 102, 69–81. [Google Scholar] [CrossRef] [Green Version]
  7. Schmitz, G.; Ecker, J. The opposing effects of n-3 and n-6 fatty acids. Prog. Lipid Res. 2008, 47, 147–155. [Google Scholar] [CrossRef] [PubMed]
  8. Kwan Tat, S.; Padrines, M.; Théoleyre, S.; Heymann, D.; Fortun, Y. IL-6, RANKL, TNF-alpha/IL-1: Interrelations in bone resorption pathophysiology. Cytokine Growth Fact. Rev. 2004, 15, 49–60. [Google Scholar]
  9. Wahba, H.M.A.; Al-Zahrany, M.S.H. Effect of feeding on diets supplemented by some vegetable oils on blood lipids and bone mineral content in osteoporotic rats. Life Sci. J. 2013, 10, 1458–1465. [Google Scholar]
  10. Hassan, H.A.; Wakf, A.M. El.; El Gharib, N.E. Role of phytoestrogenic oils in alleviating osteoporosis associated with ovariectomy in rats. Cytotechnology 2014, 66, 335–343. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  11. Longo, A.B.; Sullivan, P.J.; Peters, S.J.; LeBlanc, P.J.; Wohl, G.R.; Ward, W.E. Lifelong intake of flaxseed or menhaden oil to provide varying n-6 to n-3 PUFA ratios modulate bone microarchitecture during growth, but not after OVX in Sprague-Dawley rats. Mol. Nutr. Food Res. 2017, 61, 1600947. [Google Scholar] [CrossRef] [PubMed]
  12. Varela-López, A.; Ochoa, J.J.; Llamas-Elvira, J.M.; López-Frías, M.; Planells, E.; Speranza, L.; Battino, M.; Quiles, J.L. Loss of bone mineral density associated with age in male rats fed on sunflower oil is avoided by virgin olive oil intake or coenzyme Q supplementation. Int. J. Mol. Sci. 2017, 18, 1397. [Google Scholar] [CrossRef] [Green Version]
  13. El-Saeed, G.S.; Elghoroury, E.A.; Morsy, S.; Aly, H.M.; Wafaey, H. Phenotype of vitamin D receptor gene polymorphisms, impact of feeding flaxseed oil, and osteoporosis in ovariectomized diabetic rats. Bull. Natl. Res. Cent. 2018, 42, 11. [Google Scholar] [CrossRef] [Green Version]
  14. Elbahnasawy, A.S.; Ramzievna Valeeva, E.; El-Sayed, E.M.; Vladimirovna Stepanova, N. Protective effect of dietary oils containing omega-3 fatty acids against glucocorticoid-induced osteoporosis. J. Nutr. Health 2019, 52, 323–331. [Google Scholar] [CrossRef]
  15. Baird, H.T.; Eggett, D.; Fullmer, L S. Varying ratios of omega-6:omega-3 fatty acids on the pre and postmortem bone mineral density, bone ash, and bone breaking strength of laying chickens. Poult. Sci. 2008, 87, 323–328. [Google Scholar] [CrossRef] [PubMed]
  16. Rezq, A.A.; Labib, F.A.; Attia, A.E.M. Effect of some dietary oils and fats on serum lipid profile, calcium absorption and bone mineralization in mice. Pak. J. Nutr. 2010, 9, 643–650. [Google Scholar] [CrossRef] [Green Version]
  17. Anez-Bustillos, L.; Cowan, E.; Cubria, M.B.; VillaCamacho, J.C.; Mohamadi, A.; Dao, D.T.; Pan, A.; Fell, G.; Baker, M.A.; Nandivada, P.; et al. Effects of dietary omega-3 fatty acids on bones of healthy mice. Clin. Nutr. 2019, 38, 2145–2154. [Google Scholar] [CrossRef]
  18. Effenberger, H. The plant economy of the Northern European Bronze Age—more diversity through increased trade with southern regions. Veget. Hist. Archaeobot. 2018, 27, 65–74. [Google Scholar] [CrossRef]
  19. Martinelli, T.; Galasso, I. Phenological growth stages of Camelina sativa according to the extended BBCH scale. Ann. Appl. Biol. 2011, 158, 87–94. [Google Scholar] [CrossRef]
  20. Gugel, R.K.; Falk, K.C. Agronomic and seed quality evaluation of Camelina sativa in western Canada. Can. J. Plant Sci. 2006, 86, 1047–1058. [Google Scholar] [CrossRef]
  21. Zubr, J. Oil-seed crop: Camelina sativa. Ind. Crops Prod. 1997, 6, 113–119. [Google Scholar] [CrossRef]
  22. Abramovic, H.; Abram, V. Physiochemical properties, composition and oxidative stability of camelina sativa oil. Food Techonol. Biotech. 2005, 43, 63–70. [Google Scholar]
  23. Zubr, J.; Matthaus, B. Effects of growth conditions on fatty acids and tocopherols in Camelina sativa oil. Ind. Crops Prod. 2002, 15, 155–162. [Google Scholar] [CrossRef]
  24. Abramovic, H.; Butinar, B.; Nicolic, V. Changes occurring in phenolic content, tocopherol composition and oxidative stability of Camelina sativa oil during storage. Food Chem. 2007, 104, 903–909. [Google Scholar] [CrossRef]
  25. Waraich, E.A.; Ahmed, Z.; Ahmad, R.; Ashraf, M.Y.; Saifullah; Naeem, M.S.; Rengel, Z. Camelina sativa, a climate proof crop, has high nutritive value and multiple-uses: A review. Aust. J. Crop Sci. 2013, 7, 1551–1559. [Google Scholar]
  26. Ratusz, K.; Symoniuk, E.; Wroniak, M.; Rudzińska, M. Bioactive compounds, nutritional quality and oxidative stability of cold-pressed camelina (Camelina sativa L.) oils. Appl. Sci. 2018, 8, 2606. [Google Scholar] [CrossRef] [Green Version]
  27. Flachowsky, G.; Schaarmann, G.; Jahreis, G.; Schone, F.; Richter, G.H.; Bohme, H.; Schneider, A. Influence of feeding of oilseeds and byproducts from oilseeds on vitamin E concentration of animal products. Fett Lipid 1997, 99, 55–60. [Google Scholar] [CrossRef]
  28. Rokka, T.; Alen, K.; Valaja, J.; Ryhanen, E.L. The effect of a Camelina sativa enriched diet on the composition and sensory quality of hen eggs. Food Res. Int. 2002, 35, 253–256. [Google Scholar] [CrossRef]
  29. Peiretti, P.G.; Mussa, P.P.; Prola, L.; Meineri, G. Use of different levels of false flax (Camelina sativa L.) seed in diets for fattening rabbits. Livest. Sci. 2007, 107, 192–198. [Google Scholar] [CrossRef]
  30. Ciurescu, G.; Ropota, M.; Toncea, I.; Habeanu, M. Camelia (Camelina sativa L. Crantz Variety) oil and seeds as n-3 fatty acids rich products in broiler diets and its effects on performance, meat fatty acid composition, immune tissue weights, and plasma metabolic profile. J. Agr. Sci. Technol. 2016, 18, 315–326. [Google Scholar]
  31. Bulbul, T.; Rahman, A.; Ozdemir, V. Effect of false flax meal on certain growth, serum and meat parameters of japanese quails. J. Anim. Plant Sci. 2015, 25, 1245–1250. [Google Scholar]
  32. Puzio, I.; Jaskiewicz, T.; Sagan, A.; Bienko, M.; Grabos, D. Effects of CLA and Camelina sativa seed oil on bone properties in broiler chickens. Bull. Vet. Inst. Pulawy 2012, 56, 93–97. [Google Scholar] [CrossRef] [Green Version]
  33. Ni Eidhin, D.; Burke, J.; Lynch, B.; O’Beirne, D. Effects of dietary supplementation with camelina oil on porcine blood lipids. J. Food Sci. 2003, 68, 671–679. [Google Scholar] [CrossRef]
  34. Böhme, H.; Aulrich, K.; Schumann, W.; Fischer, K. Studies on the suitability of false flax expeller as feedstuff. 1st communication: Feeding value and incorporation limits for pigs. Fett Lipid 1997, 99, 254–259. [Google Scholar] [CrossRef]
  35. Frame, D.D.; Palmer, M.; Peterson, B. Use of Camelina sativa in the diets of young turkeys. J. Appl. Poult. Res. 2007, 16, 381–386. [Google Scholar] [CrossRef]
  36. PN-ISO 5509:1996. Analysis of Fatty Acid Methyl Esters by Gas Chromatography; The Polish Committee for Standardization: Warsaw, Poland, 1996. [Google Scholar]
  37. PN-EN ISO 5508:1996. Determination of Fatty Acid Composition; The Polish Committee for Standardization: Warsaw, Poland, 1996. [Google Scholar]
  38. Puzio, I.; Kapica, M.; Bieńko, M.; Radzki, R.P.; Pawłowska, M.; Tymicki, G. Fundectomy, antrectomy and gastrectomy influence densitometric, tomographic and mechanical bone properties as well as serum ghrelin and nesfatin-1 levels in rats. Med. Weter. 2014, 70, 604–609. [Google Scholar]
  39. Sims, N.; Morris, H.; Moore, R.; Durbridge, T.C. Increased bone resorption precedes increased bone formation in the ovariectomized rat. Calcif. Tissue Int. 1996, 59, 121–127. [Google Scholar] [CrossRef] [PubMed]
  40. Srikanta, P.; Nagarajappa, S.H.; Viswanatha, G.L.; Handral, M.; Subbanna, R.; Srinath, R.; Hiremath, G. Anti-osteoporotic activity of methanolic extract of an Indian herbal formula (NR/CAL/06) in ovariectomized rats. J. Chin. Integr. Med. 2011, 9, 1125–1132. [Google Scholar] [CrossRef]
  41. Hernandes, L.; Ramos, A.L.; Micheletti, K.R.; Santi, A.P.; Cuoghi, O.A.; Salazar, M. Densitometry, radiography, and histological assessment of collagen as methods to evaluate femoral bones in an experimental model of osteoporosis. Osteoporos. Int. 2012, 23, 467–473. [Google Scholar] [CrossRef] [PubMed]
  42. Sipos, W.; Pietschmann, P.; Rauner, M.; Kerschan-Schindl, K.; Patsch, J. Pathophysiology of osteoporosis. Wien. Med. Wochenschr. 2009, 159, 230–234. [Google Scholar] [CrossRef]
  43. Boulbaroud, S.; Mesfioui, A.; Arfaoui, A.; Ouichou, A.; el-Hessni, A. Preventive effects of flaxseed and sesame oil on bone loss in ovariectomized rats. Pak. J. Biol. Sci. 2008, 11, 1696–1701. [Google Scholar] [CrossRef] [Green Version]
  44. Dong, H.; Hutchins-Wiese, H.; Kleppinger, A.; Annis, K.; Liva, E.; Lammi-Keefe, C.; Durham, H.; Feinn, R.; MKenny, A. Effects of omega-3 polyunsaturated fatty acid supplementation on bone turnover in older women. Int. J. Vitam. Nutr. Res. 2014, 84, 124–132. [Google Scholar] [CrossRef]
  45. Griel, A.E.; Kris-Etherton, P.M.; Hilpert, K.F.; Zhao, G.; West, S.G.; Corwin, R.L. An increase in dietary n-3 fatty acids decreases a marker of bone resorption in humans. Nutr. J. 2007, 6, 2. [Google Scholar] [CrossRef] [Green Version]
  46. Rajaram, S.; Yip, E.L.; Reghunathan, R.; Mohan, S.; Sabaté, J. Effect of altering dietary n-6:n-3 polyunsaturated fatty acid ratio with plant and marine-based supplement on biomarkers of bone turnover in healthy adults. Nutrients 2017, 9, 1162. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  47. Watkins, B.A.; Reinwald, S.; Li, Y.; Seifert, M.F. Protective actions of soy isoflavones and n-3 PUFAs on bone mass in ovariectomized rats. J. Nutr. Biochem. 2005, 16, 479–488. [Google Scholar] [CrossRef]
  48. Watkins, B.A.; Li, Y.; Seifert, M.F. Dietary ratio of n-6/n-3 PUFAs and docosahexaenoic acid: Actions on bone mineral and serum biomarkers in ovariectomized rats. J. Nutr. Biochem. 2006, 17, 282–289. [Google Scholar] [CrossRef]
  49. Maggio, M.; Artoni, A.; Lauretani, F.; Ruggiero, C.; Cederholm, T.; Cherubini, A.; Ceda, G.P. The effect of polyunsaturated fatty acids on bone health. Rev. Clin. Gerontol. 2011, 21, 219–232. [Google Scholar] [CrossRef]
  50. Lau, B.Y.Y.; Cohen, D.J.A.; Ward, W.E.; Ma, D.W.L. Investigating the role of polyunsaturated fatty acids in bone development using animal models. Molecules 2013, 18, 14203–14227. [Google Scholar] [CrossRef] [Green Version]
  51. Sun, D.; Krishnan, A.; Zaman, K.; Lawrence, R.; Bhattacharya, A.; Fernandes, G. Dietary n-3 fatty acids decrease osteoclastogenesis and loss of bone mass in ovariectomized mice. J. Bone Miner. Res. 2003, 18, 1206–1216. [Google Scholar] [CrossRef] [PubMed]
  52. Watkins, B.; Li, Y.; Lippman, H.; Feng, S. Modulatory effect of omega-3 polyunsaturated fatty acids on osteoblast function and bone metabolism. Prostaglandins Leukot. Essent. Fatty Acids 2003, 68, 387–398. [Google Scholar] [CrossRef]
  53. Haag, M.; Magada, O.N.; Claassen, N.; Bohmer, L.H.; Kruger, M.C. Omega-3 fatty acids modulate ATPases involved in duodenal Ca absorption. Prostaglandins Leukot. Essent. Fatty Acids 2003, 68, 423–429. [Google Scholar] [CrossRef]
  54. Coetzer, H.; Claassen, N.; Van Papendorp, D.H.; Kruger, M.C. Calcium transport by isolated brush border and basolateral membrane vesicles: Role of essential fatty acid supplementation. Prostaglandins. Leukot. Essent. Fatty Acids 1994, 50, 257–266. [Google Scholar] [CrossRef]
  55. Armstrong, V.T.; Brzustowicz, M.R.; Wassall, S.R.; Jenski, L.J.; Stillwell, W. Rapid flip-flop in polyunsaturated docosahexaenoate) phospholipid membranes. Arch. Biochem. Biophys. 2003, 414, 74–82. [Google Scholar] [CrossRef]
  56. Valentine, R.; Valentine, D. Omega-3 fatty acids in cellular membranes: A unified concept. Prog. Lipid Res. 2004, 43, 383–402. [Google Scholar] [CrossRef]
  57. Stillwell, W.; Wassall, S.R. Docosahexaenoic acid: Membrane properties of a unique fatty acid. Chem. Phys. Lipids 2003, 126, 1–27. [Google Scholar] [CrossRef]
  58. Sepulveda, M.R.; Berrocal-Carrillo, M.B.; Gasset, M.; Mata, A.M. The plasma membrane Ca2+-ATPase isoform 4 is localized in lipid rafts of cerebellum synaptic plasma membranes. J. Biol. Chem. 2006, 281, 447–453. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  59. Priante, G.; Bordin, L.; Musacchio, E.; Clari, G.; Baggio, B. Fatty acids and cytokine mRNA expression in human osteoblastic cells: A specific effect of arachidonic acid. Clin. Sci. 2002, 102, 403–409. [Google Scholar] [CrossRef]
  60. Chandrasekar, B.; Fernandes, G. Decreased pro-inflammatory cytokines and increased antioxidant enzyme gene expression by omega-3 lipids in murine lupus nephritis. Biochem. Biophys. Res. Commun. 1994, 200, 893–898. [Google Scholar] [CrossRef] [PubMed]
  61. Bordin, L.; Priante, G.; Musacchio, E.; Giunco, S.; Tibaldi, E.; Clari, G. Arachidonic acid-induced IL-6 expression is mediated by PKC alpha activation in osteoblastic cells. Biochemistry 2003, 42, 4485–4491. [Google Scholar] [CrossRef] [PubMed]
  62. Ferrucci, L.; Cherubini, A.; Bandinelli, S.; Bartali, B.; Corsi, A.; Lauretani, F.; Martin, A.; Andres-Lacueva, C.; Senin, U.; Guralnik, J.M. Relationship of plasma polyunsaturated fatty acids to circulating inflammatory markers. J. Clin. Endocrinol. Metab. 2006, 91, 439–446. [Google Scholar] [CrossRef] [PubMed]
  63. Caughey, G.E.; Mantzioris, E.; Gibson, R.A.; Cleland, L.G.; James, M.J. The effect on human tumor necrosis factor alpha and interleukin 1 beta production of diets enriched in n-3 fatty acids from vegetable oil or fish oil. Am. J. Clin. Nutr. 1996, 63, 116–122. [Google Scholar] [CrossRef]
  64. Bagga, D.; Wang, L.; Farias-Eisner, R.; Glaspy, J.A.; Reddy, S.T. Differential effects of prostaglandin derived from omega-6 and omega-3 polyunsaturated fatty acids on COX-2 expression and IL-6 secretion. Proc. Natl. Acad. Sci. USA 2003, 100, 1751–1756. [Google Scholar] [CrossRef] [Green Version]
  65. Calder, P.C. N-3 Polyunsaturated fatty acids, inflammation, and inflammatory diseases. Am. J. Clin. Nutr. 2006, 83, 1505S–1519S. [Google Scholar] [CrossRef]
  66. Lacey, D.L.; Grosso, L.E.; Moser, S.A.; Erdmann, J.; Tan, H.L.; Pacifici, R.; Villareal, D.T. IL-1-induced murine osteoblast IL-6 production is mediated by the type 1 IL-1 receptor and is increased by 1,25 dihydroxyvitamin D3. J. Clin. Investig. 1993, 91, 1731–1742. [Google Scholar] [CrossRef] [Green Version]
  67. Collins, D.A.; Chambers, T.J. Effect of prostaglandins E1, E2 and F2a on osteoclast formation in mouse bone marrow cultures. J. Bone Miner. Res. 1991, 6, 157–164. [Google Scholar] [CrossRef]
  68. Jee, W.S.S.; Ma, Y.F. The in vivo anabolic actions of prostaglandins in bone. Bone 1997, 21, 297–304. [Google Scholar] [CrossRef]
  69. Watkins, B.A.; Li, Y.; Allen, K.G.; Hoffmann, W.E.; Seifert, M.F. Dietary ratio of (n-6)/(n-3) polyunsaturated fatty acids alters the fatty acid composition of bone compartments and biomarkers of bone formation in rats. J. Nutr. 2000, 130, 2274–2284. [Google Scholar] [CrossRef]
  70. Gao, Q.; Xu, M.; Alander, C.B.; Choudhary, S.; Pilbeam, C.C.; Raisz, L.G. Effects of prostaglandin E2 on bone in mice in vivo. Prostaglandins Other Lipid Mediat. 2009, 89, 20–25. [Google Scholar] [CrossRef] [PubMed]
  71. Watkins, B.A.; Shen, C.L.; Mcmurtry, J.P.; Xu, H.; Bain, S.D.; Allen, K.G.D.; Seifert, M.F. Dietary lipids modulate bone prostaglandin E2 production, insulin-like growth factor-I concentration and formation rate in chicks. J. Nutr. 1997, 127, 1084–1091. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  72. Mollard, R.C.; Gillam, M.E.; Wood, T.M.; Taylor, C.G.; Weiler, H.A. (n-3) fatty acids reduce the release of prostaglandin E2 from bone but do not affect bone mass in obese (fa/fa) and lean Zucker rats. J. Nutr. 2005, 135, 499–504. [Google Scholar] [CrossRef] [PubMed]
  73. Al-Nouri, D.M.; El-Din, M.F.S.; Al-Khalifa, A.S. The effect of long-term supplementation with different dietary ω-6/ω-3 ratios on mineral content and ex vivo prostaglandin E2 release in bone of growing rabbits. Nutr. Res. Pract. 2014, 8, 360–367. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  74. Al-Nouri, D.M.; Al-Khalifa, A.S.; Shahidi, F. Long-term supplementation of dietary omega-6/omega-3 ratios alters bone marrow fatty acid and biomarkers of bone metabolism in growing rabbits. J. Funct. Foods 2012, 4, 584–593. [Google Scholar] [CrossRef]
  75. Hamid, R.; Singh, J.; Reddy, B.S.; Cohen, L.A. Inhibition by dietary menhaden oil of cyclooxygenase-1 and 2 in N-nitrosomethylurea-induced rat mammary tumors. Int. J. Oncol. 1999, 14, 523–528. [Google Scholar] [CrossRef] [PubMed]
  76. Liu, D.; Denbow, D.M. Maternal dietary lipids modify composition of bone lipids and ex vivo prostaglandin E2 production in early postnatal Japanese quail. Poult. Sci. 2001, 80, 1344–1352. [Google Scholar] [CrossRef] [PubMed]
  77. Raisz, L.G.; Alander, C.B.; Simmons, H.A. Effects of prostaglandin E3 and eicosapentaenoic acid on rat bone in organ culture. Prostaglandins 1989, 37, 615–625. [Google Scholar] [CrossRef]
  78. Zhong, X.; Gao, S.; Wang, J.J.; Dong, L.; Huang, J.; Zhang, L.L.; Wang, T. Effects of linseed oil and palm oil on growth performance, tibia fatty acid and biomarkers of bone metabolism in broilers. Br. Poult. Sci. 2014, 55, 335–342. [Google Scholar] [CrossRef] [PubMed]
  79. Corwin, R.L. Effects of dietary fats on bone health in advanced age. Prostaglandins Leukot. Essent. Fatty Acids 2003, 68, 379–386. [Google Scholar] [CrossRef]
  80. Tian, X.Y.; Zhang, Q.; Setterberg, R.B.; Zeng, Q.Q.; Iturria, S.J.; Ma, Y.F. Continuous PGE2 leads to net bone loss while intermittent PGE2 leads to net bone gain in lumbar vertebral bodies of adult female rats. Bone 2008, 42, 914–920. [Google Scholar] [CrossRef]
  81. Liu, X.H.; Kirschenbaum, A.; Yao, S.; Levine, A.C. Interactive effect of interleukin-6 and prostaglandin E2 on osteoclastogenesis via the OPG/RANKL/RANK system. Ann. N. Y. Acad. Sci. 2006, 1068, 225–233. [Google Scholar] [CrossRef]
  82. Coetzee, M.; Haag, M.; Kruger, M.C. Effects of arachidonic acid, docosahexaenoic acid, prostaglandin E(2) and parathyroid hormone on osteoprotegerin and RANKL secretion by MC3T3-E1 osteoblast-like cells. J. Nutr. Biochem. 2007, 18, 54–63. [Google Scholar] [CrossRef]
  83. Udagawa, N.; Koide, M.; Nakamura, M.; Nakamichi, Y.; Yamashita, T.; Uehara, S.; Kobayashi, Y.; Furuya, Y.; Yasuda, H.; Fukuda, C.; et al. Osteoclast diferentiation by RANKL and OPG signaling pathways. J. Bone Miner. Metab. 2021, 39, 19–26. [Google Scholar] [CrossRef]
  84. Wright, H.L.; McCarthy, H.S.; Middleton, J.; Marshall, M.J. RANK, RANKL and osteoprotegerin in bone biology and disease. Curr. Rev. Musculoskelet. Med. 2009, 2, 56–64. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  85. Ono, K.; Kaneko, H.; Choudhary, S.; Pilbeam, C.C.; Lorenzo, J.A.; Akatsu, T. Biphasic effect of prostaglandin E2 on osteoclast formation in spleen cell cultures: Role of the EP2 receptor. J. Bone Miner. Res. 2005, 20, 23–29. [Google Scholar] [CrossRef] [PubMed]
  86. Tsutsumi, R.; Xie, C.; Wei, X.; Zhang, M.; Zhang, X.; Flick, L.M. PGE2 signaling through the EP4 receptor on fibroblasts upregulates RANKL and stimulates autolysis. J. Bone Miner. Res. 2009, 24, 1753–1762. [Google Scholar] [CrossRef]
  87. Kakhki, R.A.M.; Shouldice, V.L.; Price, K.R.; Moats, J.; Kiarie, E.G. n-3 fatty acids fed to ISA brown and Shaver white breeders and their female progeny during rearing: Impact on egg production, eggshell, and select bone attributes from 18 to 42 weeks of age. Poult. Sci. 2020, 99, 3959–3970. [Google Scholar] [CrossRef] [PubMed]
  88. Sobol, M.; Skiba, G.; Raj, S. Dietary n-3 PUFA content as a modulator of the femur properties in growing pigs. Br. J. Nutr. 2019, 121, 508–518. [Google Scholar] [CrossRef] [PubMed]
  89. Weiler, H.A.; Fitzpatrick-Wong, S.C. Modulation of essential (n-6):(n-3) fatty acid ratios alters fatty acid status but not bone mass in piglets. J. Nutr. 2002, 132, 2667–2672. [Google Scholar] [CrossRef] [Green Version]
  90. Abdulla, N.R.; Loha, T.C.; Akit, H.; Sazili, A.Q.; Fooe, H.L.; Kareem, K.Y.; Mohamad, R.; Rahim, R.A. Effects of dietary oil sources, calcium and phosphorus levels on growth performance, carcass characteristics and bone quality of broiler chickens. J. Appl. Anim. Res. 2017, 45, 423–429. [Google Scholar] [CrossRef]
Table
Table 1. Fatty acid composition of camelina oil. The contents of particular fatty acids are presented as a percentage of total FAME.
Table 1. Fatty acid composition of camelina oil. The contents of particular fatty acids are presented as a percentage of total FAME.
Fatty AcidsContent
C14:00.05
C15:00.02
C16:05.12
C16:10.13
C17:00.06
C18:02.42
C18:114.94
C18:217.42
C18:333.95
C20:01.35
C20:114.35
C20:21.44
C20:41.03
C22:00.3
C22:12.91
C24:00.18
C24:10.67
Total n-6 PUFA19.89
Total n-3 PUFA33.95
Table
Table 2. Final body weight, parameters of body composition, and densitometric and mechanical properties of femur.
Table 2. Final body weight, parameters of body composition, and densitometric and mechanical properties of femur.
ParametersSHOOVX-PhSOVX-CO5OVX-CO9
BW (g)295 ± 2 a305 ± 6324 ± 7 b324 ± 10 b
LM (g)213 ± 5 a258 ± 8 b264 ± 7 b247 ± 6 b
FM (g)23.8 ± 2.4 a36.6 ± 5.847.1 ± 7,058.0 ± 11.9 b
Tot.BMC (g)8.58 ± 0.20 a7.55 ± 0.11 b8.49 ± 0.24 a8.94 ± 0.35 a
Tot.BMD (g/cm2)0.143 ± 0.003 a0.133 ± 0.002 b0.137 ± 0.0010.136 ± 0.001
Femur
weight (g)0.78 ± 0.05 a0.77 ± 0.04 a0.85 ± 0.06 b0.89 ± 0.08 b
length (mm)34.9 ± 0.0534.8 ± 0.0635.3 ± 0.0335.1 ± 0.06
fBMD (g/cm2)0.094 ± 0.002 a0.091 ± 0.001 b0.098 ± 0.002 a0.096 ± 0.001 a
fBMC (g)0.336 ± 0.010 a0.301 ± 0.006 b0.341 ± 0.008 a0.338 ± 0.007 a
Ultimate force (N)111.3 ± 7.3 a96.4 ± 7.1 a176.9 ± 13.2 b157.8 ± 12.3 b
Work to fracture (mJ)14.35 ± 2.08 ac10.86 ± 2.77 a37.75 ± 6.33 b29.82 ± 4.94 bc
Young’s modulus (GPa)3.38 ± 0.152.79 ± 0.133.43 ± 0.393.03 ± 0.34
The results are means ± S.E.M (n = 10). The values in the rows are significantly different (p ≤ 0.05) as indicated by the superscripts: a, b, c. Abbreviations: SHO —shame operated rats receiving physiological saline; OVX-PhS —ovariectomized rats receiving physiological saline; OVX-CO5 —ovariectomized rats receiving camelina oil in the amount of 5 g/kg/BW; OVX-CO9 —ovariectomized rats receiving camelina oil in the amount of 9 g/kg/BW; BW—body weight; LM—lean mass; FM—fat mass; Tot. BMC—bone mineral content of total skeleton; Tot. BMD—bone mineral density of total skeleton; fBMD—bone mineral density of femur; fBMC—bone mineral content of femur.
Table
Table 3. Tomographic analysis (pQCT) of bone tissue in the middle part of femur diaphysis.
Table 3. Tomographic analysis (pQCT) of bone tissue in the middle part of femur diaphysis.
ParametersSHOOVX-PhSOVX-CO5OVX-CO9
T.vBMD (mg/mm3)805 ± 6 a764 ± 9 b852 ± 7 c827 ± 6 ac
T.BMC (mg/mm)7.30 ± 0.117.15 ± 0.04 a7.74 ± 0.19 b7.81 ± 0.18 b
Ct.vBMD (mg/mm3)1420 ± 41419 ± 31424 ± 51430 ± 4
Ct.BMC (mg/mm)6.83 ± 0.12 a6.74 ± 0.06 a7.19 ± 0.177.32 ± 0.14 b
T.Ar (mm2)9.09 ± 0.189.08 ± 0.199.31 ± 0.239.69 ± 0.17
Ct.Ar (mm2)4.73 ± 0.074.77 ± 0.06 a5.13 ± 0.11 b5.11 ± 0.09 b
Ct.Th (mm)0.527 ± 0.003 a0.522 ± 0.004 a0.569 ± 0.011 b0.558 ± 0.006 b
Peri.C (mm)10.7 ± 0.110.7 ± 0.110.8 ± 0.110.9 ± 0.1
Endo.C (mm)7.41 ± 0.106.89 ± 0.647.37 ± 0.157.46 ± 0.10
PMI (mm4)10.35 ± 0.3710.17 ± 0.3311.24 ± 0.4511.58 ± 0.42
SSI (mm3)5.67 ± 0.125.45 ± 0.10 5.97 ± 0.196.17 ± 0.22
The results are means ± S.E.M (n = 10). The values in the rows are significantly different (p ≤ 0.05) as indicated by the superscripts: a, b, c. Abbreviations: T.vBMD—total volumetric bone mineral density; T.BMC—total bone mineral content; Ct.vBMD—cortical volumetric bone mineral density; Ct.BMC—cortical bone mineral content; T.Ar—total bone area; Ct.Ar—cortical bone area; Ct.Th—cortical thickness; Peri.C—periosteal circumferences; Endo.C—endocortical circumferences; PMI—polar moment of inertia of the cortical bone; SSI—strength-strain index.
Table
Table 4. Biochemical markers of bone metabolism.
Table 4. Biochemical markers of bone metabolism.
ParametersSHOOVX-PhSOVX-CO5OVX-CO9
Osteocalcin (ng/mL)129.7 ± 8.4 a246.9 ± 14.8 b378.2 ± 25.1 c356.7 ± 23.0 c
CTX-I (ng/mL)18.37 ± 1.95 a30.01 ± 2.11 b22.13 ± 1.68 a22.14 ± 1.74 a
IL-6 (pg/mL)280.3 ± 26.2 a457.2 ± 53.7 b307.5 ± 32.1292.7 ± 39.9
The results are means ± S.E.M (n = 10). The values in the rows are significantly different (p ≤ 0.05) as indicated by the superscripts: a, b, c. Abbreviations: CTX-1—C-terminal telopeptide of type I collagen; IL-6—interleukin-6.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Puzio, I.; Graboś, D.; Bieńko, M.; Radzki, R.P.; Nowakiewicz, A.; Kosior-Korzecka, U. Camelina Oil Supplementation Improves Bone Parameters in Ovariectomized Rats. Animals 2021, 11, 1343. https://doi.org/10.3390/ani11051343

AMA Style

Puzio I, Graboś D, Bieńko M, Radzki RP, Nowakiewicz A, Kosior-Korzecka U. Camelina Oil Supplementation Improves Bone Parameters in Ovariectomized Rats. Animals. 2021; 11(5):1343. https://doi.org/10.3390/ani11051343

Chicago/Turabian Style

Puzio, Iwona, Dorota Graboś, Marek Bieńko, Radosław P. Radzki, Aneta Nowakiewicz, and Urszula Kosior-Korzecka. 2021. "Camelina Oil Supplementation Improves Bone Parameters in Ovariectomized Rats" Animals 11, no. 5: 1343. https://doi.org/10.3390/ani11051343

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop