Home Medicine Investigation of the relationship cellular and physiological degeneration in the mandible with AQP1 and AQP3 membrane proteins
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Investigation of the relationship cellular and physiological degeneration in the mandible with AQP1 and AQP3 membrane proteins

  • Mustafa Çiçek , Velid Unsal ORCID logo EMAIL logo and Mehmet Kemal Tümer
Published/Copyright: January 23, 2020
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Turkish Journal of Biochemistry
From the journal Turkish Journal of Biochemistry Volume 45 Issue 5

Abstract

Objective

In this study, we aimed to investigate the changes in the levels of oxidative stress and antioxidant enzymes on the mandibular bone caused by the expression of aquaporin-1 and aquaporin-3 proteins.

Material and method

14 Balb/C white mice were divided into two groups of seven, based on whether they are young or old. Mandibular tissue samples were taken for biochemical and histological analysis.

Results

Findings of our study has shown that, AQP-1 and AQP-3 immunoreactivity significantly decreased in mandibular bone tissues of aged mice in comparison to younger mice (p < 0.05). MDA and AOPP levels, which are the indicators of oxidative stress, increased in elderly mice and antioxidant defense system SOD enzyme activity was decreased (p < 0.05). The TNF-α cytokine level, which is the indicator for inflammations, was found to be higher in older mice than in young mice (p < 0.05).

Conclusion

As a result, it was observed that cellular damage, disruption in water – electrolyte balance and increased inflammation that occur during the natural process of aging had caused serious and irreversible disturbances.

Öz

Amaç

Bu çalışmada, aquaporin-1 ve aquaporin-3 proteinlerinin ekspresyonunun neden olduğu mandibular kemik üzerindeki oksidatif stres seviyelerindeki ve antioksidan enzim aktivitelerindeki değişiklikleri araştırmayı amaçladık.

Gereç ve Yöntem

14 Balb/C beyaz fare genç veya yaşlı olup olmadıklarına göre yedişer iki gruba ayrıldı. Biyokimyasal, histolojik analiz için mandibular doku örnekleri alındı.

Bulgular

Çalışmamızın bulguları, AQP-1 ve AQP-3 immünoreaktivitesinin, yaşlı farelerin mandibular kemik dokularında, genç farelere kıyasla anlamlı olarak azaldığını göstermiştir (p < 0.05). Oksidatif stresin göstergesi olan MDA ve AOPP düzeyleri yaşlı farelerde artmış ve antioksidan savunma sistemi SOD enzim aktivitesi düşmüştür (p < 0.05). Enflamasyonun göstergesi olan TNF-α sitokin seviyesinin yaşlı farelerde genç farelere göre daha yüksek olduğu bulunmuştur. (p < 0.05).

Sonuç

Doğal yaşlanma sürecinde meydana gelen hücresel hasar, su – elektrolit dengesinde bozulma ve artmış enflamasyonun ciddi ve geri dönüşü olmayan bozukluklara neden olduğu görülmüştür.

Keywords: Aquaporin-1; Aquaporin-3; Oxidative stress; TNF-alpha
Anahtar Kelimeler: Aquaporin-1; Aquaporin-3; Oksidatif Stres; TNF-alfa

Introduction

Mandibular bone, fossa mandibulare in temporal bone and temporomandibular joint (TMJ) are all part of the stomatognathic system [1]. Among the most important changes that aging has on the organism, the changes that occur in the oral cavity and changes that occur in the tissues that surround the cavity are among the most significant [2]. With aging, calcified tissue in all bones, often in women, decrease, which results in porosity and increases the brittleness of the bones. Blood flow in both maxilla and mandible is also decreased in elderly. [3]. Mandibular subluxations or dislocations occur as a result of trauma more often in elderly individuals and fractures develop more often due to thinned chondyle structure [4]. The volume and osmolarity of the cell is maintained by the carrier properties of the membrane-embedded proteins. The plasma membrane acts as a barrier for the passage of water and the special water channels (aquaporins) were identified for the first time in cells with high water permeability [5]. Thirteen different types of aquaporin (AQP) families were identified in mammals and these channels also aid in the transportation of glycerol, urea, arsenite and some ions. Aquaporins that pass through the membrane 6 times do not allow the passage of protons, however, they allow the passage of 5–9 water molecules through the channels they form through the special amino acid sequences they contain [6].

In this study, we have aimed to show the biochemical changes, expressions of aquaporin 1 and aquaporin 3 and the histopathology and immunohistochemical changes that occur in the of mandibular bone, which is a part of temporomandibular joint component, due to aging.

Materials and methods

Procurement of animals

After obtaining the permission of the local ethics committee (2015-HADYEK-28), this study was carried out in experimental medicine research unit of Gaziosmanpaşa University. In the study 14 Balb/C species white mice were used. Animals were divided into two equal groups of seven. Group 1 included young animals which were 2 months old (n=7) and Group 2 included old animals which were 18 months old (n=7). Prior to application of the experiment, the mice were kept at room temperature (22±1°C) and 40–50% humidity. The light pattern was adjusted to 12 h of day and 12 h of night. Eating and drinking habits of the animals were left untouched. The rats were kept under observation for a week and their daily physical examinations were carried out.

Taking samples

Mice without any health problems were put down using the excretion technique while they were under an anesthesia of ketamine/xylazine (50/10 mg/kg). Mandibulae and a portion of the temporal bone of the mice were removed in a manner which would sustain the bone integrity and these were placed inside a 10% EDTA solution for the purposes of biochemical, immunohistochemical and histopathological examinations.

Histological examination of mandibular bone

Tissues were kept in the EDTA solution until histological sections could be obtained. After the routine histological follow-up, the tissues were embedded in paraffin. 4–5 Hm thick sections were taken from paraffin-embedded tissues and they were stained with hematoxylin-eosin (H&E) method. The stained sections were examined under Euromex Oxion light microscope.

Immunohistochemistry

The 4–5 μm sections taken from the paraffin blocks were placed on polylysine slides. After dehydrating the deparaffinized tissues using graded alcohol series, the tissues placed inside distilled water and they were boiled for 5 min in the microwave oven (600 W) at pH: 6 in citrate buffer solution for antigen retrieval. To prevent endogenous peroxidase activity, they were treated with H2O2. To prevent surface staining, primary antibody was incubated for 60 min using (Rabbit polyclonal to Aquaporin 1, Abcam, AB110186, Cambridge, UK; Rabbit polyclonal to Aquaporin 3, Abcam, AB34710, Cambridge, UK) after the application of Ultra V Block (Ultra V Block, TA-125-UB, Thermo Fisher Scientific Inc., Waltham, MA, USA) solution. After the application of the primary antibody (biotinylated anti-mouse IgG, Diagnostic BioSystems, KP 50A, Pleasanton, CA, USA) was administered to the secondary antibody for 30 min, streptavidin horseradish peroxidase was administered for another 30 min and 3-amino-9-ethyl carbazole chromogen was applied and contrast painting was carried out using Mayers hematoxylin. In the tissues prepared for negative control, phosphate buffered saline (PBS) was used instead of primary antibody, and other steps were performed in the same manner. The tissues that were put through PBS and distilled water were sealed using an appropriate sealing solution. The prepared preparates were examined and photographed using the research microscope (Euromex Oxion). Evaluation of immunohistochemical markers was performed using H-SCORE analyses [7].

Immunoreactivity intensity of AQP-1 and AQP-3 were evaluated using half-quantitative methods through the tracked intensity categories: 0 (no staining), 1+ (weak but detectable stain), 2+ (medium or apparent staining), and 3+ (intense staining). For each tissue, an H-score value was obtained by first calculating the percentages of cells according to the intensity category of their staining (AQP-1 and AQP-3). After that, this stain weighted value was multiplied by=ΣPi (i l+) using the H-SCORE value in which “i” represents the intensity values and “Pi” represents the related percentage of cells. All slides were evaluated under light microscope (×40 magnification). The percentage of cells in each density within these areas was determined by two researchers at different times, who were not aware of the type and source of tissues. The mean score of both observers was used.

Tissue sampling, homogenate preparation and biochemical measurements

Biochemical analyzes

The bone tissue separated for biochemical analysis was first divided into two with pliers. SOD, MDA, AOPP were examined with the first part of the bone, TNF-α, sodium, and potassium were examined with the second part. Each piece was weighted and homogenized separately. First, it was continued to be cut into small pieces with pliers. The crushed or comminuted bone tissue was then homogenized in mortar. To produce 10% (w/v) homogenate, bone were homogenized with ice-cold Tris-HCl buffer (0.15M, pH 7.4) for 10 min by automatic tissue homogenizer (Ultra Turrax Type T 18, IKA Labortechnic, Germany). Then the homogenate was centrifuged at 7.000×g for 15min. The pellet was separated and the clear supernatant was used for the analysis. SOD, MDA, AOPP were measured from the obtained supernatants [8], [9]. Bone tissue was demineralized in 0.5 M EDTA, pH=8 (4 mL/g) and incubated at room temperature overnight with rocking. Samples were centrifuged at 6000 rcf for 15 min and supernatants were collected. Pellets were resuspended in an additional 2 mL/g 0.5 M EDTA, incubated at room temperature for 5 days with rocking, and then centrifuged. EDTA supernatants were collected and pooled with the previous EDTA fraction. Pellets were resuspended in 2 mL/g of 6 M guanidine hydrochloride in 0.1 M Tris (pH 7.4) and incubated at 65 C on a heating block overnight. Samples were centrifuged and supernatants were collected. The solubilization step was repeated twice more (again with 2 mL/g for 1 day, then with 4 mL/g for 2 days), and all three guanidine hydrochloride supernatants were combined (total final volume 8 mL/g). Pellets were then discarded [10]. The supernatants obtained from Na+ and K+ bone were examined using indirect ion selective electrode (ISE) method and TNF-α was examined through the ELISA method using a SUNRED brand commercial set. All procedures were performed at 4°C.

Protein levels

Concentration of the protein was determined according to the procedure described by Lowry et al. [11].

Assay of AOPP and MDA levels

Similar to the previous studies, AOPP levels were evaluated with the 200 μL supernatant samples, which were diluted in the phosphate buffered saline in a proportion of 1:5 using spectrophotometric methods (readings at 340 nm) [12].

Similar to the previous studies, 200 μL s supernatant samples were mixed with 1000 mL tris-HCl buffer and 300 mL KH2PO4 for the evaluation of the MDA levels. The reaction was stopped with 1 mL 10% trichloroacetic acid after the incubation. Following the warming of samples in the boiling water for 20 min, after adding 1500 mL thiobarbituric acid, the samples were read with the spectrophotometric methods at 532 nm [13].

Determination of SOD activity

SOD activity was determined with the measurement with the spectrophotometric methods at 560 nm according to the inhibition principle of the nitro blue tetrazolium (NBT) with the xanthine-xanthine oxidase system, which was used as the superoxide generator. This method was described by Sun et al. [14].

Statistical analysis

Normality control was performed using the Shapiro-Wilk test. The data was found to be suitable for normal distribution. The level of statistical significance was taken as p<0.05. Two independent sample t tests were used to compare the groups. While investigating associations of data, correlation coefficients and their significance were calculated with Spearman’s test (for non-normally distributed variables) and Pearson’s test (for normally distributed variables).

Results

Biochemical findings

Na+, K+, TNF-α values were determined by the ELISA measurements carried out on supernatants they are as provided with the Table 1. MDA and AOPP levels, which are indicators for cell damage and lipid peroxidation in tissues, were found to be higher in elderly mice compared to young mice (p<0.05). In comparison to the younger mice, a significant decrease in SOD enzyme activity in elderly mice was observed in antioxidant defense system (p<0.05). K+ levels found in bone tissue and bone metabolism was determined to be higher in young mice in comparison to the elderly mice and it was determined that their Na+ levels were lower (p<0.05). TNF-α level, which is a intracellular inflammatory cytokine, was significantly increased in elderly mice compared to young mice (p<0.05) (Figure 1).

Table 1:

Results of biochemical analysis of mandible bone tissues of young and old mice.

Results of biochemical analysis of mandible
Group
ParameterYoung miceOld micep-Value
x̄±±SDx̄±SD
TNF-α (ng/mg protein)99.47±8.80171.74±15.41 (+)0.001
MDA (nmol/mg protein)5.07±0.638.13±0.82 (+)0.001
AOPP (μmol/mg protein)93.62±13.57139.72±27.44 (+)0.001
SOD (U/mg protein)82.94±7.3156.97±5.82 (−)0.001
Sodium (Eq/g protein)49.54±4.7440.28±4.34 (−)0.03
Potassium (Eq/g protein)1.62±0.282.38±0.33 (+)0.02

  1. Values are expressed as x̄±SD (n=7). TNF-α, Tumor necrosis factor alpha; MDA, Malondialdehyde; AOPP, Advanced oxidation protein products; SOD, Superoxide dismutase.

Figure 1: Graphical illustration of biochemistry results of mandible bone tissue in young and old mouse.
Figure 1:

Graphical illustration of biochemistry results of mandible bone tissue in young and old mouse.

While analyzing correlations, we found significant and correlations between TNF-α and SOD activity (r=−0.594, p<0.028), MDA, and AOPP levels (r=0.638, p<0.021; r=0.575, p<0.029) (Table 2).

Table 2:

Correlations of SOD, MDA, AOPP with TNF-α, sodium, potassium.

SODMDAAOPP
rp-Valuerp-Valuerp-Value
TNF-α−0.5940.0480.6380.0210.5750.029
Sodium0.2130.57−0.3790.401−0.430.244
Potassium−0.3930.320.3020.4030.2530.512

  1. Those with significant p-values are marked in bold.

Histopathological findings

In all the sections examined, it has been observed that the mandibular bone is composed of proliferation, chondrogenic and hypertrophic segments. Increased thickness of proliferative, chondrogenic and hypertrophic layers have been observed in young mice. Irregularly distributed mature chondrocytes with large, spherical shapes were observed in the hypertrophic region of the older mice, whereas in younger mice, these cells were found to be more numerous and in a specific order. It has been observed that the bone marrow of the mandibular bone is more numerous in young mice and the boundaries of spongious form trabeculae are more pronounced in older mice. In young mice, blood vessels were observed with ease and in older mice there was a decrease in the count blood vessels and these vessels had structural deformations and expansions in arterioles and veins. A significant amount of cartilage deterioration was observed in aged mice. Bone tissue and connective tissue around the bone were observed in all mice, young and old. Substantia compacta layer of younger mice was found to be tighter and morphometrically larger compared to that of the older mice. In young mice, fibrous connective tissue formed a broader band than that of the aged mice. In young mice, numerous hypertrophic chondrocytes were found in the transition area from bone tissue to articular cartilage. In young mice, trabecular bone structure has been observed in almost all areas except the articular cartilage area of the epiphysis (Figure 2).

Figure 2: Hematoxylin–eosin staining, 40×.Mandible bone tissue (A) young mice (B) old mice (yellow arrow; blood vessels, black arrow; osteoblasts, green arrow; compact bone layer of mandible, blue arrow; osteocytes).
Figure 2:

Hematoxylin–eosin staining, 40×.

Mandible bone tissue (A) young mice (B) old mice (yellow arrow; blood vessels, black arrow; osteoblasts, green arrow; compact bone layer of mandible, blue arrow; osteocytes).

Immunohistochemical findings

AQP-1 and AQP-3 immunoreactivity

AQP-1 and AQP-3 proteins were stained immunohistochemically in the mandibular bone tissue tests of both the old and young mice. The results were evaluated semi-quantitatively and they are shown on the Table 3. AQP-1 and AQP-3 immunoreactivity of young mice in mandibular bone tissues were significantly increased (+++) (Figure 3) and a low level of (+) (p=0.001) staining was observed in the AQP and AQP-3 immunoreactivity of the elderly mice (Figure 3). The H-score results of immunopositive staining of osteoblast cells in the substantia compact and substantia spongiosa regions of the mandibular bone showed significantly higher AQP-1 and AQP-3 immunoreactivity in young mice than in older mice (p=0.001) (Table 4).

Table 3:

Aquaporin-1 and aquaporin-3 immunoreactivity of the young and old mice.

Intensity of immunoreactivity staining
Group
ParameterYoung miceOld mice
IntensitynIntensitynp-Value
Mandibulae AQP-1+++7+70.001a
Mandibulae AQP-3+++7+70.001a

  1. The results were evaluated semiquantitatively. AQP-1, Aquaporin-1; AQP-3, Aquaporin-3.

Figure 3: The structures indicated by arrows are aquaporin protein positive stained cells.Immunohistochemical staining of aquaporin-1 and aquaporin-3, 40× (A) aquaporin-1 young mice (B) aquaporin-1 old mice (C) aquaporin-3 young mice (D) aquaporin-3 old mice.
Figure 3:

The structures indicated by arrows are aquaporin protein positive stained cells.

Immunohistochemical staining of aquaporin-1 and aquaporin-3, 40× (A) aquaporin-1 young mice (B) aquaporin-1 old mice (C) aquaporin-3 young mice (D) aquaporin-3 old mice.

Table 4:

Aquaporin-1 and aquaporin-3 H-score comparisons of the young and old mice.

Comparison of H-score values
Group
ParameterYoung miceOld micetp-Value
x̄±SDnx̄±SDn
Mandibulae AQP-1159.47±20.2137106.35±6.21376.180.001a
Mandibulae AQP-3148.71±18.427100.17±1.1275.840.001a

  1. Values are expressed as x̄±SD (n=7). AQP-1, Aquaporin-1; AQP-3, Aquaporin-3.

Discussion

Mandibular bone has a significant role in the masticatory system. Since morphological deterioration of its structure causes chewing problems and teeth problems in both humans and animals, this matter possesses a clinical significance [15]. Some in vitro studies have shown that the maximum force during chewing is exerted on the lateral and anterior surface of the condyle of the mandibular bone [16]. In this study, biochemical, histopathological and immunohistochemical changes in bone tissue were evaluated in order to better understand the damage caused by aging on the mandibular bone tissue. The literature review has revealed that our study is the most extensive research on the effects that aging has on the mandibular bone tissue. The sodium concentration difference between intracellular fluid and intercellular fluid is a result of active transport by Na+-K+ ATPase [17]. One study reported that physiological processes of aging is associated with changes in water metabolism and sodium balance and it causes volumetric changes in plasma osmolality and body fluid levels [18]. Studies have shown that osteoclast-mediated bone resorption occurs to protect sodium homeostasis. According to this hypothesis, the Na+, absorbed from the bones through the detection of low Na+ content in the body by the osteoclasts through low extracellular fluid levels is retained by the kidney by activation of the renin-angiotensin-aldosterone system [19].

Mandibular bone tissue homogenization analysis of our study has revealed a significant decrease in Na+ levels which occurred in parallel with aging. It is our belief that Na+ content of the bone tissue is low due to irreversible nature of osteoclast activity caused by aging. Through the Na+ K+ ATPase activity, K+ ions are kept in high concentration within these cells and this ion, together with proteins, has an active role in the regulation of the osmotic pressure and fluid volume, and also in maintaining the acid-base balance of the cell [20]. Bone buffer system is very important in chronic acidosis and bone resorption leads to Na+, K+, HCO3− and Ca++ loss from bone. A recent study has indicated that in the cells with Na+/K+-ATPase canal loss, there may be large amounts of K+ translocation from tissues that has suffered from ischemia or necrosis on the extracellular area [21].

Since the water retention capacity of the cells is decreased and cellular damage level is increased with aging, the amount of K+ in bone tissue significantly higher in older mice compared to young mice. It has been suggested that the mechanism underlying the initiation and progression of the aging process is oxidative stress [22]. Neutrophil derivatives are defined as ditrosin, which are advanced oxidation protein products (AOPP) formed by reaction between chlorinated oxidants and proteins that contain protein cross-linker products. The increased AOPP levels indicate the oxidative damage on the proteins [23].

A study, which examined the relationship between oxidative stress and age-related bone loss has determined that AOPP levels in plasma and femur tissue increased with aging. In a previous study, the same researchers have demonstrated that AOPP can inhibit the osteogenic differentiation of osteoblast-like cells [24]. When the AOPP levels in bone tissue were examined in the results of our study, it was determined that older mice had higher AOPP levels than younger ones. MDA is seen as one of the final products of ROS-induced lipid peroxidation and it is a parameter of oxidative stress [25]. In a recent clinical study, plasma MDA levels were observed to be higher in postmenopausal women with osteoporosis [26].

According to the findings of our study, tissue MDA levels that we thought to be caused by aging and the cellular damage caused by aging increased significantly in aged mice. SOD is one of the antioxidant enzymes and it can prevent superoxide accumulation by the conversion of superoxide to hydrogen peroxide [25]. An experiment carried on rats have revealed that that SOD activity of older rats was lower than that of young and mature rats [24]. Findings of our study were similar. According to our findings, SOD activity of older mice were significantly lower than young mice. Based on our data on oxidative stress and antioxidant activity, we can say that the aging process causes the accumulation of oxidative modified products and the degradation of the antioxidant defense system. TNF-α promotes bone absorption by stimulating the proliferation and differentiation of osteoclasts or by activating mature osteoclasts [27]. Some studies have revealed that as a result of estrogen deficiency, osteoclastogenesis synthesis increases, which in turn increases the production of TNF-α [28]. In a study on fracture healing, whether the increase in expression of TNF-α in osteoporosis was related to upregulation of osteoclasts was examined. As a result, it was found that up-regulation of osteoplasty was correlated with increase of TNF-α [29].

A similar situation was also noted in our study. TNF-α levels of elderly mice were increased significantly compared to young mice. We believe that this is caused by the up-regulated expression of TNF-α, which increases the number of osteoclasts due to aging.

Many factors play a role in growth and development of the mandible such as diet, testosterone, estrogen, gender, age and genetics [30]. In one study, changes in the cartilages of the mandibular bone and changes in the subchondral bone were simultaneously analyzed and it was determined that in young mice, age and gender based changes in the size of subchondral bone and mandibular bone were significant [31]. According to the histopathological results of our study, proliferative, chondrogenic and hypertrophic layers of the young mice were found to be thicker than the older mice. In aged mice, chondrocyte cells in the hypertrophic layer were found to be more irregular and smaller in number than in young mice. Compared with aged mice, the mandibular bone marrow of young mice was greater in number and the boundaries of trabeculae in the substantia spongiosa layer showed a less pronounced but more orderly lay-out. A significant number of pronounced blood vessels were observed in young mice. In aged mice, there was a serious degradation in the connective tissue around periosteum and around bone and cartilage tissue. The substantia compacta layer of younger mice was thicker, tighter and larger. Aquaporins provide the transportation of water and some small soluble substances through the cell membrane. It is expressed in many tissues and has important functions in the body’s fluid homeostasis [32]. Water and oxygen can easily pass through the AQP-1 channels and these channels facilitate the diffusion of oxygen from the plasma membranes. It also prevents rapid volume deformations in cases of osmotic stress [33]. In a study on the pathogenesis of osteoarthritis, an increased expression of AQP-1 was observed and it was believed that this played a role in the pathogenesis of osteoarthritis [34].

In one study, the presence of AQP-1 was expressed in bone tissues of both young and old rats. This proves that, both in young and old mice, AQP 1 and 3 are physiologically involved in the discs of the joint [35]. In our study, positive immunoreactivity of AQP-1 in mandibular bone tissue of young and old mice was demonstrated. In the light of these results, the presence of AQP-1 in mandibular bone tissue, which is a component of the TMJ, demonstrates that it has a physiological role in the joint disc. Especially in young mice, intense staining of bone marrow, endosteum, osteoblasts in trabeculae, osteoclasts and osteocytes were observed. Clinical data obtained from a study has determined a high incidence rate of AQP-1 in osteosarcoma tissues and it has been reported that AQP-1 can be used as a useful prognostic marker and potential target in the treatment of osteosarcoma cancer. In the same study, the researches have predicted that AQP-1 plays a key role in the proliferation, apoptosis and metastasis of osteosarcoma cells, and that AQP-1 can regulate this through TGF-b and focal adhesion signalling [36]. In the data of our study, H-Score analysis of AQP-1 has revealed that young mice had high AQP-1 cell count and that this cell count was decreased significantly in elderly mice. We believe that in young mice there is an intensive substance exchange between the extracellular material and the intracellular environment through AQP-1 channels in cell membrane and in elderly mice, due to the increase in cellular degradation and the natural process of aging, water channels tasked with substance exchange may decrease in number, which causes homeostasis function to deteriorate. AQP-3 has been proven to be present in many tissues such as kidney, respiratory epithelium, choroid plexus, endometrium and subchondral osteoblasts [33]. In addition to its normal physiological functions, AQP-3 is thought to play a role in the pathophysiology of some diseases. Increased expression of AQP-3 mRNA in different regions of osteoarthritis cartilage and the presence of AQP-3 proteins suggest that it has important functions in the pathogenesis of osteoarthritis [37]. In our study, the positive immunoreactivity of AQP-3 in the mandibular bone tissue of young and old mice proves the presence of AQP-3 in bone tissue. The expression of AQP-3 in tissues transforms with aging [35]. Increased arginine and vasopressin levels are common in the elderly and these increased levels can cause water retention and hypoanthremia and may lead increased risk of fracture by causing osteoporosis through stimulating excess calcium release from the bones [38].

In our study, H-Score analyses revealed that young mice had a high AQP-3 cell counts, which decreased significantly in older mice. AQP-3 immunoreactivity was found to denser in older mice compared to older mice. When the decreases in Na+ increases of MDA levels and K+, which are the indicators of cell damage, were considered in our study, it was determined that, the degradation of the water-electrolyte balance caused by expressions of AQP-1 and AQP-3 increases the risk of hyponatremia, osteoporosis and bone tissue fracture.

Conclusion

As a result of this, mandibular bone tissue is weakened with aging and material strength of bone is also decreased. We think that this is caused by the expansion of the internal medullary cavity and the increase in the number and size of the Hawers channels. Aging related bone loss occurs largely due to increased osteoclastic activity. Since all animals we used in our experiments were female, we attributed the increase of osteoclastic activity to the reduction of ovarian hormones after menopause, which is the natural process of aging, degradation in water and electrolyte balance, decrease in number of aquaporin water channels in the cell membrane and morphological, physiological and functional deformations.

Acknowledgments

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article. The author(s) received no financial support for the research, authorship, and/or publication of this article.

  1. Conflict of interest: Authors have no conflict of interest.

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Received: 2019-05-06
Accepted: 2019-12-25
Published Online: 2020-01-23

©2020 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Review Article
  3. Newly developed diagnostic methods for SARS-CoV-2 detection
  4. Short Communication
  5. Effect of hemolysis on prealbumin assay
  6. Research Articles
  7. BioVar: an online biological variation analysis tool
  8. High dose ascorbic acid treatment in COVID-19 patients raised some problems in clinical chemistry testing
  9. Immunoassay biomarkers of first and second trimesters: a comparison between pregnant Syrian refugees and Turkish women
  10. Association of maternal serum trace elements with newborn screening-thyroid stimulating hormone
  11. PIK3CA and TP53 MUTATIONS and SALL4, PTEN and PIK3R1 GENE EXPRESSION LEVELS in BREAST CANCER
  12. Evaluation of E2F3 and survivin expression in peripheral blood as potential diagnostic markers of prostate cancer
  13. Age, gender and season dependent 25(OH)D levels in children and adults living in Istanbul
  14. Original Article
  15. Fractional excretion of magnesium as an early indicator of renal tubular damage in normotensive diabetic nephropathy
  16. Research Articles
  17. Diagnostic value of laboratory results in children with acute appendicitis
  18. Evaluation of thiol disulphide levels in patients with pulmonary embolism
  19. Relationship between renal tubulointerstitial fibrosis and serum prolidase enzyme activity
  20. Comparison of test results obtained from lithium heparin gel tubes and serum gel tubes
  21. MHC Class I related chain A (MICA), Human Leukocyte Antigen (HLA)-DRB1, HLA-DQB1 genotypes in Turkish patients with ulcerative colitis
  22. An overview of procalcitonin in Crimean-Congo hemorrhagic fever: clinical diagnosis, follow-up, prognosis and survival rates
  23. Comparison of different equations for estimation of low-density lipoprotein (LDL) – cholesterol
  24. Case-Report
  25. A rare case of fructose-1,6-bisphosphatase deficiency: a delayed diagnosis story
  26. Research Articles
  27. Atypical cells in sysmex UN automated urine particle analyzer: a case report and pitfalls for future studies
  28. Investigation of the relationship cellular and physiological degeneration in the mandible with AQP1 and AQP3 membrane proteins
  29. In vitro assessment of food-derived-glucose bioaccessibility and bioavailability in bicameral cell culture system
  30. Letter to the Editor
  31. The weighting factor of exponentially weighted moving average chart
https://doi.org/10.1515/tjb-2019-0174
Keywords for this article
Aquaporin-1; Aquaporin-3; Oxidative stress; TNF-alpha

Articles in the same Issue

  1. Frontmatter
  2. Review Article
  3. Newly developed diagnostic methods for SARS-CoV-2 detection
  4. Short Communication
  5. Effect of hemolysis on prealbumin assay
  6. Research Articles
  7. BioVar: an online biological variation analysis tool
  8. High dose ascorbic acid treatment in COVID-19 patients raised some problems in clinical chemistry testing
  9. Immunoassay biomarkers of first and second trimesters: a comparison between pregnant Syrian refugees and Turkish women
  10. Association of maternal serum trace elements with newborn screening-thyroid stimulating hormone
  11. PIK3CA and TP53 MUTATIONS and SALL4, PTEN and PIK3R1 GENE EXPRESSION LEVELS in BREAST CANCER
  12. Evaluation of E2F3 and survivin expression in peripheral blood as potential diagnostic markers of prostate cancer
  13. Age, gender and season dependent 25(OH)D levels in children and adults living in Istanbul
  14. Original Article
  15. Fractional excretion of magnesium as an early indicator of renal tubular damage in normotensive diabetic nephropathy
  16. Research Articles
  17. Diagnostic value of laboratory results in children with acute appendicitis
  18. Evaluation of thiol disulphide levels in patients with pulmonary embolism
  19. Relationship between renal tubulointerstitial fibrosis and serum prolidase enzyme activity
  20. Comparison of test results obtained from lithium heparin gel tubes and serum gel tubes
  21. MHC Class I related chain A (MICA), Human Leukocyte Antigen (HLA)-DRB1, HLA-DQB1 genotypes in Turkish patients with ulcerative colitis
  22. An overview of procalcitonin in Crimean-Congo hemorrhagic fever: clinical diagnosis, follow-up, prognosis and survival rates
  23. Comparison of different equations for estimation of low-density lipoprotein (LDL) – cholesterol
  24. Case-Report
  25. A rare case of fructose-1,6-bisphosphatase deficiency: a delayed diagnosis story
  26. Research Articles
  27. Atypical cells in sysmex UN automated urine particle analyzer: a case report and pitfalls for future studies
  28. Investigation of the relationship cellular and physiological degeneration in the mandible with AQP1 and AQP3 membrane proteins
  29. In vitro assessment of food-derived-glucose bioaccessibility and bioavailability in bicameral cell culture system
  30. Letter to the Editor
  31. The weighting factor of exponentially weighted moving average chart
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