Effects of Electroacupuncture on Bone Mass and Cathepsin K Expression in Ovariectomised Rats

Objective To characterise the effects of early and late electroacupuncture (EA) treatment on serum 17β-oestradiol (E2), C-terminal cross-linking telopeptide of type I collagen (CTX-I), bone mineral density (BMD), biomechanical bone strength and mRNA expression of cathepsin K in ovariectomised (OVX) rats. Methods Sixty Sprague-Dawley rats underwent ovariectomy (n=40) or sham surgery (n=20) and were randomly divided into two batches. Batch 1 (n=30) consisted of 10 sham-operated rats (Sham-0 group) and 20 OVX rats: half commenced EA immediately (early EA group, n=10) and half were left untreated (OVX-0 group, n=10). Batch 2 (n=30) consisted of 10 sham-operated rats (Sham-12 group) and 20 OVX rats: half commenced EA treatment 12 weeks after ovariectomy (late EA group, n=10) and half were left untreated (OVX-12 group, n=10). Rats in batches 1 and 2 were killed after 12 and 24 weeks, respectively. Serum E2, CTX-I, BMD, bone strength and cathepsin K expression were determined by ELISA, dual energy X-ray absorptiometry, biomechanical assessment and qRT-PCR, respectively. Results Both early and late EA treatment increased serum E2 levels, reduced serum CTX- I levels and increased BMD and bone strength of the L5 vertebral body in OVX rats. Although early EA treatment similarly increased BMD and bone strength of the femur, late EA treatment did not. However, both early and late EA treatment reduced mRNA expression of cathepsin K in OVX rats. Conclusions Early EA completely prevented and late EA partially prevented bone loss and deterioration of bone strength in OVX rats. The timing of initiation of EA treatment may be an important consideration for optimisation of effects. The influence of EA on bone strength appears to be at least partially mediated through regulation of the expression of cathepsin K.

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The Preventive Effect of Biochanin A on Bone Loss in Ovariectomized Rats: Involvement in Regulation of Growth and Activity of Osteoblasts and Osteoclasts

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2013/594857 ◽

2013 ◽

Vol 2013 ◽

pp. 1-10 ◽

Cited By ~ 7

Author(s):

Shu-Jem Su ◽

Yao-Tsung Yeh ◽

Huey-Wen Shyu

Keyword(s):

Bone Loss ◽

Mrna Expression ◽

Biological Effects ◽

Biochanin A ◽

Ovariectomized Rats ◽

Tartrate Resistant Acid Phosphatase ◽

Type I ◽

Mineral Density ◽

Ovx Rats

Biochanin A (BCA) is a major isoflavone abundant in red clover (Trifolium pretense). The protective effect of BCA on bone loss in an ovariectomized (OVX) animal model has never been clarified. The objective of this study was to investigate the biological effects of BCA on bone loss in OVX ratsin vivoand on the development of osteoblasts and osteoclastsin vitro. Ovariectomy resulted in a marked increase in body weight and a decrease in femoral bone mineral density and trabecular bone volume that was prevented by BCA or 17β-estradiol (E2) treatment. However, an increase in uterine weight was observed in E2-treated OVX rats, but not in response to BCA treatment. Treatment with BCA increased the mRNA expression of osterix, collagen type I, alkaline phosphatase (ALP), and osteocalcin and decreased the mRNA expression of tartrate-resistant acid phosphatase (TRAP) and the receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) ratio in the femur of OVX rats. Treatment with BCA or E2 prevented the OVX-induced increase in urinary deoxypyridinoline (DPD) and serum tumor necrosis factorα(TNF-α) and interleukin-1β(IL-1β).In vitro, BCA induced preosteoblasts to differentiate into osteoblasts and increased osteoblast mineralization. BCA inhibited preosteoclasts and osteoclast proliferation and decreased osteoclast bone resorption. These findings suggest that BCA treatment can effectively prevent the OVX-induced increase in bone loss and bone turnover possibly by increasing osteoblastic activities and decreasing osteoclastic activities.

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Prenylated Isoflavonoids-Rich Extract of Erythrinae Cortex Exerted Bone Protective Effects by Modulating Gut Microbial Compositions and Metabolites in Ovariectomized Rats

Nutrients ◽

10.3390/nu13092943 ◽

2021 ◽

Vol 13 (9) ◽

pp. 2943

Author(s):

Hui-Hui Xiao ◽

Xueli Yu ◽

Chen Yang ◽

Chi-On Chan ◽

Lu Lu ◽

...

Keyword(s):

Bone Microarchitecture ◽

Cathepsin K ◽

Short Chain Fatty Acids ◽

Female Rats ◽

Ovariectomized Rats ◽

Protective Effects ◽

Sprague Dawley ◽

Mineral Density ◽

Beneficial Effects ◽

Ovx Rats

Flavonoids, found in a wide variety of foods and plants, are considered to play an important role in the prevention and treatment of osteoporosis. Our previous studies demonstrated that Erythrina cortex extract (EC) rich in prenylated isoflavonoids exerted bone protective effects in ovariectomized (OVX) rats. The present study aimed to investigate the interactions of gut microbiota with the EC extract to explore the underlying mechanisms involved in its beneficial effects on bone. Sprague-Dawley female rats of 3-months-old were ovariectomized and treated with EC extract for 12 weeks. EC extract reversed ovariectomy-induced deterioration of bone mineral density and bone microarchitecture as well as downregulated cathepsin K (Ctsk) and upregulated runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP) in the tibia of OVX rats. Its protective effects on bone were correlated with changes in microbial richness and the restorations of several genera. EC increased the serum circulating levels of acetate and propionate in OVX rats. We conclude that the bone protective effects of EC extract were associated with the changes in microbial compositions and serum short chain fatty acids (SCFAs) in OVX rats.

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Prolonged feeding of difructose anhydride III increases strength and mineral concentrations of the femur in ovariectomized rats

British Journal Of Nutrition ◽

10.1079/bjn20051483 ◽

2005 ◽

Vol 94 (2) ◽

pp. 268-274 ◽

Cited By ~ 19

Author(s):

Rieko Mitamura ◽

Hiroshi Hara

Keyword(s):

Bone Strength ◽

Ovariectomized Rats ◽

Control Group ◽

Sprague Dawley ◽

Mineral Density ◽

Ovx Rats ◽

Sprague Dawley Rats ◽

Promotive Effect ◽

Mineral Concentrations ◽

The Relationship

This study demonstrates that feeding difructose anhydride III (DFAIII) improves bone strength and femoral mineral concentrations in a rat model of oestrogen deficiency. We showed the relationship between Ca, Mg and P absorption and bone characteristics in rats. Two groups of female Sprague-Dawley rats (6 weeks old) underwent bilateral ovariectomy (ovariectomized rats, OVX rats) or bilateral laparotomy (sham rats). At 10 weeks old, OVX and sham rats were divided into three subgroups and fed a control, 1·5 % DFAIII or 3 % DFAIII diet for 8 weeks, respectively. Ca but not Mg absorption rates were lowered by ovariectomy; however, ingestion of the 1·5 % and 3 % DFAIII diets similarly restored the reduced Ca absorption in OVX rats at 4 and 8 weeks after feeding of the test diets. DFAIII increased Mg absorption dose-dependently in sham and OVX rats. The bone strength, femoral Ca and Mg concentrations, and distal bone mineral density in the 3 % DFAIII group were higher than those in the control group in OVX rats. The absorption rates of Ca and Mg were significantly correlated with femoral Ca and Mg concentrations and strength, which suggests that increasing both Ca and Mg absorption improves bone characteristics in OVX rats. There were no differences in any of the variables in the femur between the 1·5 % and 3 % DFAIII groups in OVX rats. In conclusion, feeding of a low dose of DFAIII increased intestinal Ca and Mg absorption, and the promotive effect of DFAIII persisted for over 8 weeks. This effect was associated with prevention of ovariectomy-induced osteopenia.

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In Men With Obesity, T2DM Is Associated With Poor Trabecular Microarchitecture and Bone Strength, and Low Bone Turnover

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/clinem/dgab061 ◽

2021 ◽

Author(s):

Francesca Vigevano ◽

Giulia Gregori ◽

Georgia Colleluori ◽

Rui Chen ◽

Vimlin Autemrongsawat ◽

...

Keyword(s):

Bone Turnover ◽

Bone Strength ◽

Type I Collagen ◽

Failure Load ◽

Bone Microarchitecture ◽

Enzyme Linked Immunosorbent Assay ◽

Type I ◽

Mineral Density ◽

Trabecular Microarchitecture ◽

Increased Risk

Abstract Introduction Obesity and type 2 Diabetes (T2D) are both associated with greater bone mineral density (BMD) but increased risk of fractures. The effect of the combination of both conditions on bone metabolism, microarchitecture and strength in the obese population remains unknown. Methods Data from 112 obese men were collected. Bone turnover and biochemical markers were measured by enzyme-linked immunosorbent assay (ELISA), body composition and BMD at all sites were assessed by dual energy X-ray absorptiometry (DXA), whereas bone microarchitecture and strength (stiffness and failure load) were measured by high-resolution peripheral computed tomography (HR-pQCT). Data were compared among metabolically healthy obese (MHO) and metabolically unhealthy obese (MUHO) with and without T2D and between obese without and with T2D. Results Compared to MHO and MUHO without T2D, MUHO with T2D had significantly lower levels of osteocalcin ((7.49±3.0 and 6.03±2.47, vs 4.24±2.72 ng/ml, respectively, p=0.003) and C-terminal telopeptide of type I collagen (CTx) (0.28±0.10 and 0.29±0.13 vs. 0.21±0.15 ng/ml, respectively, p=0.02). Dividing our subjects simply into those with and without T2D showed that obese men with T2D had significantly lower levels of osteocalcin (p=0.003) and CTx (p=0.005), greater trabecular separation at the tibia and radius (p=0.03 and p=0.04, respectively) and lower tibial failure load and stiffness (both p=0.04), relative to obese men without T2D. Conclusion In men, the combination of obesity and T2D is associated with reduced bone turnover, poorer trabecular bone microarchitecture and bone strength compared to those who are obese but without T2D suggesting worse bone disease.

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Association of Bone-Related Biomarkers with Femoral Neck Bone Strength

10.21203/rs.3.rs-854913/v1 ◽

2021 ◽

Author(s):

Ning Xia ◽

Yun Cai ◽

Wei Wang ◽

Chen Bao ◽

Yunming Li ◽

...

Keyword(s):

Bone Loss ◽

Femoral Neck ◽

Bone Strength ◽

Type I Collagen ◽

Linear Regression Analysis ◽

Fragility Fractures ◽

Maximum Load ◽

Multiple Linear Regression Analysis ◽

Type I ◽

Mineral Density

Abstract Background: Femoral neck fractures are the worst consequence of osteoporosis (OP), and its early prevention and treatment have become a public health problem.This study aims to investigate the relationship of bone-related biomarkers,femoral neck bone mineral density (BMD) and maximum load (Lmax),selecting the indicator which can reflect femoral neck bone loss and reduced bone strength. Methods: A total of 108 patients were recruited from January 2017 to December 2019. Femoral neck BMD, femoral neck Lmaxand bone-related biomarkerswere all measured and analyzed.Results: The expression of femoral neck type I collagen (COL-I)were significantly decreased, whereas other markers were all significantly increased with the decreasing of femoral neck BMD and Lmax(P<0.05). Among them, serum C-terminal telopeptide of type I collagen (CTX)levels and femoral neck osteopontin (OPN) expression were increased in osteopenia. In multiple linear regression analysis,CTX and OPN were independent factorsof femoral neck BMD and Lmax, whereasCOL-I was independent factor affectingLmax(P< 0.05). Moreover, CTX was the independent risk factor of OPN and COL-I(P<0.05).Conclusions: Bone-related biomarkersdo reflect decreased femoral neck bone mass and bone strength. Of them, the CTX increases most significantly in the process of bone loss, and can be used as a surrogate markerof OPN and COL-I for bone loss and reduced bone strength.Early measurement of CTXcouldfacilitate the diagnosis of osteopenia and provide a theoretical basis for delaying the occurrence of femoral neck OP and fragility fractures.

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Cells in regenerating deer antler cartilage provide a microenvironment that supports osteoclast differentiation

Journal of Experimental Biology ◽

10.1242/jeb.204.3.443 ◽

2001 ◽

Vol 204 (3) ◽

pp. 443-455

Author(s):

C. Faucheux ◽

S. Nesbitt ◽

M. Horton ◽

J. Price

Keyword(s):

Type I Collagen ◽

Mononuclear Cells ◽

Osteoclast Differentiation ◽

Cathepsin K ◽

Collagen Matrix ◽

Tartrate Resistant Acid Phosphatase ◽

Type I ◽

Deer Antler

Deer antlers are a rare example of mammalian epimorphic regeneration. Each year, the antlers re-grow by a modified endochondral ossification process that involves extensive remodelling of cartilage by osteoclasts. This study identified regenerating antler cartilage as a site of osteoclastogenesis in vivo. An in vitro model was then developed to study antler osteoclast differentiation. Cultured as a high-density micromass, cells from non-mineralised cartilage supported the differentiation of large numbers of osteoclast-like multinucleated cells (MNCs) in the absence of factors normally required for osteoclastogenesis. After 48 h of culture, tartrate-resistant acid phosphatase (TRAP)-positive mononuclear cells (osteoclast precursors) were visible, and by day 14 a large number of TRAP-positive MNCs had formed (783+/−200 per well, mean +/− s.e.m., N=4). Reverse transcriptase/polymerase chain reaction (RT-PCR) showed that receptor activator of NF κ B ligand (RANKL) and macrophage colony stimulating factor (M-CSF) mRNAs were expressed in micromass cultures. Antler MNCs have the phenotype of osteoclasts from mammalian bone; they expressed TRAP, vitronectin and calcitonin receptors and, when cultured on dentine, formed F-actin rings and large resorption pits. When cultured on glass, antler MNCs appeared to digest the matrix of the micromass and endocytose type I collagen. Matrix metalloproteinase-9 (MMP-9) may play a role in the resorption of this non-mineralised matrix since it is highly expressed in 100 % of MNCs. In contrast, cathepsin K, another enzyme expressed in osteoclasts from bone, is only highly expressed in resorbing MNCs cultured on dentine. This study identifies the deer antler as a valuable model that can be used to study the differentiation and function of osteoclasts in adult regenerating mineralised tissues.

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(deleted)

Bulletin of Russian State Medical University ◽

10.24075/10.24075/brsmu.2021.003 ◽

2021 ◽

Author(s):

MV Osikov ◽

EV Davydova ◽

KS Abramov

Keyword(s):

Bone Healing ◽

Standard Therapy ◽

Type I Collagen ◽

Femoral Shaft ◽

Control Group ◽

Blood Levels ◽

Type I ◽

Mineral Density ◽

Fracture Consolidation ◽

To Receive

Efferent physical therapy holds promise as an adjunct to the combination treatment of femoral fractures in young, working-age individuals. The aim of the study was to investigate the dynamics of bone turnover markers at different stages of femoral fracture consolidation in patients undergoing ozone therapy. The study enrolled 20 men (group 2, 47.8 ± 3.5 years) with a femoral shaft fracture (AO/ASIF 32А, 32В). The control group (group 1, 46.8 ± 3.7 years) comprised 10 healthy males. Subgroup 2a (n = 10) was assigned to receive standard therapy; subgroup 2b (n = 10) was assigned to receive standard therapy complemented by minor autohemotherapy (MAHT) at 20 mg/L ozone concentrations. On days 7, 30 and 90, fracture consolidation was assessed on the RUST scale and blood levels of С-terminal telopeptides of type I collagen (bCTx, pg/ml) and procollagen type I carboxy-terminal propeptide (PICP, ng/ml) were measured. On day 7, the total RUST score in subgroups 2a and 2b was 4 points; on day 30, it was 6.5 and 8.7 points, respectively, and on day 90, it reached 10 and 11.5 points, respectively. Bone mineral density was as high as 90% in the MAHT subgroup vs. 78% in subgroup 2а, indicating faster bone healing. On day 30, bCTx levels in subgroup 2b were higher than in subgroup 2a (2289.4 [2145.3; 2365.4] vs. 1894.6 [1745.3; 2098.2], respectively. On day 7, PICP was significantly elevated in subgroup 2b in comparison with subgroup 2a; its levels peaked on days 30 and 90 (day 30: 268.3 [231.2; 286.3] vs. 183.2 [174.6; 195.6]; day 90: 584.6 [512.3; 589.3] vs. 351.2 [312.3; 369.4]. Thus, MAHT produces a positive effect on the quality and intensity of bone healing in men with isolated closed femoral shaft fractures.

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Pharmacological inhibitors to identify roles of cathepsin K in cell-based studies: a comparison of available tools

Biological Chemistry ◽

10.1515/bc.2009.092 ◽

2009 ◽

Vol 390 (9) ◽

Cited By ~ 36

Author(s):

Sylvie Desmarais ◽

Frédéric Massé ◽

M. David Percival

Keyword(s):

Human Cell ◽

Type I Collagen ◽

Cathepsin K ◽

The Other ◽

Enzyme Assays ◽

Type I ◽

Treatment Of Osteoporosis ◽

Cysteine Cathepsins ◽

Rodent Cell ◽

Enzyme Selectivity

Abstract Cathepsin K (Cat K) degrades bone type I collagen and is a target for the pharmacological treatment of osteoporosis. Further roles for Cat K have been recently described, some of which are supported by the use of purportedly selective Cat K inhibitors in human and rodent cell-based assays. Twelve commercial and non-commercial Cat K inhibitors were profiled against a panel of purified human, rat, and mouse cysteine cathepsins and in two cell-based enzyme occupancy assays for activity against Cat K, B, and L. Ten inhibitors, including the carbohydrazide Cat K inhibitor II (Boc-Phe-Leu-NHNH-CO-NHNH-Leu-Z), the non-covalent K4b, and the epoxide NC-2300, have either little Cat K selectivity, or appear poorly cell penetrant. The amino-acetonitrile-containing inhibitors L-873724 and odanacatib show greater than 100-fold human Cat K enzyme selectivity and have similar IC50 values against each cathepsin in cell-based and enzyme assays. The basic inhibitor balicatib has greater cellular potencies than expected on the basis of purified enzyme assays. The accumulation of [14C]-balicatib in fibroblasts is blocked by prior treatment of the cells with NH4Cl, consistent with balicatib having lysosomotropic properties. These results support the use of L-873724 and odanacatib as tools to identify novel roles for Cat K using human cell-based systems, but suggest using caution in the interpretation of studies employing the other compounds.

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Electroacupuncture Ameliorates Subchondral Bone Deterioration and Inhibits Cartilage Degeneration in Ovariectomised Rats

Acupuncture in Medicine ◽

10.1136/acupmed-2016-011258 ◽

2018 ◽

Vol 36 (1) ◽

pp. 37-43 ◽

Cited By ~ 3

Author(s):

Jun Zhou ◽

Peirui Zhong ◽

Ying Liao ◽

Jing Liu ◽

Yuan Liao ◽

...

Keyword(s):

Subchondral Bone ◽

Type I Collagen ◽

Cartilage Degeneration ◽

Cross Linking ◽

Control Group ◽

Sham Operation ◽

Type I ◽

Trabecular Thickness ◽

Ovx Rats ◽

Time Point

Objectives To investigate the effects of electroacupuncture (EA) on subchondral bone mass and cartilage degeneration in an experimental animal model of osteoarthritis (OA) induced by ovariectomy (OVX). Methods Ninety 3-month-old female Sprague-Dawley rats were randomly divided into the following three groups (n = 30 each): sham operation without treatment (control group); OVX without treatment (OVX group);, and ovariectomy with EA treatment (EA group). Rats in the EA group received EA treatment from the day of OVX. Ten rats in each group were randomly killed at 4, 8 and 12 weeks after operation. Results EA reduced urine C-terminal cross-linking telopeptide of type I collagen from 4 weeks after OVX, reduced C-terminal cross-linking telopeptide of type II collagen and body weight from 8 weeks after OVX, and increased serum 17β-oestradiol from 4 weeks after OVX compared with the OVX group (all p<0.01). In the EA group, trabecular bone volume ratio, trabecular thickness and trabecular number increased, and trabecular separation were reduced at each time point compared with the OVX group (p<0.05, p<0.01, respectively). In the EA group, osteoprotegerin (OPG) expression was increased and receptor activator of nuclear factor kappa-B ligand (RANKL) expression was reduced at each time point compared with the OVX group (p<0.05, p<0.01, respectively). Mankin scores and mRNA expression of matrix metalloproteinase-13 (MMP-13) were lower in EA versus OVX groups at 12 weeks after OVX (both p<0.01). Conclusion The results suggest that EA inhibits subchondral bone loss by regulating RANK/RANKL/OPG signalling and protects articular cartilage by inhibiting MMP-13 in OVX rats.

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