Collagen Extract Derived from Yeonsan Ogye Chicken Increases Bone Microarchitecture by Suppressing the RANKL/OPG Ratio via the JNK Signaling Pathway.

Yeonsan Ogye is a traditional Korean chicken breed (Gallus domesticus, GD), with a dominant gene for fibromelanosis, showing entirely black fluffy head feathers, ear lobes, and pupils. GD collagen extract (78.6 g per 100 g total protein) was derived from the flesh of Yeonsan Ogye. The effects of GD collagen on bone mass, microarchitecture, osteogenic, osteoclastogenic differentiations, and function factor expression were investigated in ovariectomized (OVX) rats. GD collagen stimulated osteogenesis in OVX rats and increased tibial bone strength and calcium content. Micro-computed tomography analysis of tibia cross-sections revealed that GD collagen attenuated the OVX-induced changes in trabecular thickness, spacing, and number. GD collagen stimulated alkaline phosphatase activity, bone-specific matrix proteins (alkaline phosphatase (ALP), osteocalcin, collagen type I (COL-I)) and mineralization by activating bone morphogenetic protein 2 (BMP-2)/mothers against decapentaplegic homolog 5 (SMAD5)/runt-related transcription factor 2 (Runx2). GD collagen inhibited osteoclast differentiation and function gene markers (TRAP, cathepsin K) by interfering with the Wnt signaling, increasing OPG production, and reducing the expression of RANKL, TRAP, and cathepsin K. GD collagen promoted osteogenesis by activating the p38 signal pathway and prevented osteoclastogenesis by lowering the RANKL/OPG ratio and blocking the JNK signaling pathway. Dietary supplementation with GD collagen might inhibit osteoclastogenesis, stimulate osteoblastogenesis, and regulate bone metabolism.

Vitamin C Activates Osteoblastogenesis and Inhibits Osteoclastogenesis via Wnt/ß-Catenin/ATF4 Signaling Pathways.

This study evaluated the effects of vitamin C on osteogenic differentiation and osteoclast formation, and the effects of vitamin C concentration on bone microstructure in ovariectomized (OVX) Wistar rats. Micro-computed tomography analysis revealed the recovery of bone mineral density and bone separation in OVX rats treated with vitamin C. Histomorphometrical analysis revealed improvements in the number of osteoblasts, osteoclasts, and osteocytes; the osteoblast and osteoclast surface per bone surface; and bone volume in vitamin C-treated OVX rats. The vitamin C-treated group additionally displayed an increase in the expression of osteoblast differentiation genes, including bone morphogenetic protein-2, small mothers against decapentaplegic 1/5/8, runt-related transcription factor 2, osteocalcin, and type I collagen. Vitamin C reduced the expression of osteoclast differentiation genes, such as receptor activator of nuclear factor kappa-B, receptor activator of nuclear factor kappa-B ligand, tartrate-resistant acid phosphatase, and cathepsin K. This study is the first to show that vitamin C can inhibit osteoporosis by promoting osteoblast formation and blocking osteoclastogenesis through the activation of wingless-type MMTV integration site family/ß-catenin/activating transcription factor 4 signaling, which is achieved through the serine/threonine kinase and mitogen-activated protein kinase signaling pathways. Therefore, our results suggest that vitamin C improves bone regeneration.

Image of the Micro-Computed Tomography and Atomic-Force Microscopy of Bone in Osteoporosis Animal Model.

Osteoporosis, a major public health problem, is characterised by a reduction in and deterioration of bone tissue microarchitecture, with a consequent increase in bone frailty and susceptibility to fractures. We performed ex vivo micro-computed tomography (micro-CT) and atomic-force microscopy (AFM) between normal and surgically ovariectomized animals (OVX). The tibial architecture was investigated using maximum intensity projection images. Micro-CT analysis of the tibia metaphysis was used to calculate the trabecular spacing (Tb.Sp). Following OVX surgery, the rats exhibited increased Tb.Sp. Between the bundles, there are holes in the fabric with a diameter in the range of 3 to 5 µm in normal tibia. The diameter of hole in the osteoporosis tibia is the range of 5 to 10 µm, and its area increased than normal tibia. AFM can be used for the inspection of bone surfaces and offers the possibility of quantification of local geometries, such as pit size, collagen fibril diameters, or collagen ultrastructure. Our results suggest that AFM will surely become one of the most important tools in the realm of medical research for imaging such as bone and tissues.

Optimal extent of prophylactic central neck dissection for papillary thyroid carcinoma: Comparison of unilateral versus bilateral central neck dissection.

OBJECTIVE: The aim of the study was to determine the optimal extent of prophylactic central neck dissection (pCND) in papillary thyroid carcinoma (PTC). MATERIALS AND METHODS: We studied 384 patients with clinically node-negative unilateral PTC who had undergone total thyroidectomy with pCND. Of these, 169 patients underwent unilateral pCND, and 215, bilateral pCND. RESULTS: Age, sex, and TNM stage did not differ between the two groups. The rates of occult central lymph node metastasis were 34.3% and 37.2% in the unilateral and bilateral pCND groups, respectively (p = 0.558). Metastasis to the contralateral paratracheal lymph node occurred in 4.2% of the bilateral pCND group. Major complication rates did not differ between the two groups, except for transient hypoparathyroidism, which was higher in the bilateral group (43.7% vs. 33.7%, p = 0.047). Stimulated thyroglobulin and RAI ablation uptake rates were similar in the two groups, as were recurrence and disease-free survival. CONCLUSION: Contralateral paratracheal node dissection may be not indicated for prophylactic central neck dissection in clinically node-negative unilateral PTC because the rate of contralateral paratracheal node metastasis is low and transient hypoparathyroidism is higher following bilateral dissection.

Melanin extract from Gallus gallus domesticus promotes proliferation and differentiation of osteoblastic MG-63 cells via bone morphogenetic protein-2 signaling.

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of 50-250 µg/mL. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to 250 µg/mL and were 149% and 129% at 250 µg/mL concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of 500 µg/mL. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

Calcium Supplement Derived from Gallus gallus domesticus Promotes BMP-2/RUNX2/SMAD5 and Suppresses TRAP/RANK Expression through MAPK Signaling Activation.

The present study evaluated the effects of a calcium (Ca) supplement derived from Gallus gallus domesticus (GD) on breaking force, microarchitecture, osteogenic differentiation and osteoclast differentiation factor expression in vivo in Ca-deficient ovariectomized (OVX) rats. One percent of Ca supplement significantly improved Ca content and bone strength of the tibia. In micro-computed tomography analysis, 1% Ca supplement attenuated OVX- and low Ca-associated changes in bone mineral density, trabecular thickness, spacing and number. Moreover, 1% Ca-supplemented diet increased the expression of osteoblast differentiation marker genes, such as bone morphogenetic protein-2, Wnt3a, small mothers against decapentaplegic 1/5/8, runt-related transcription factor 2, osteocalcin and collagenase-1, while it decreased the expression of osteoclast differentiation genes, such as thrombospondin-related anonymous protein, cathepsin K and receptor activator of nuclear factor kappa B. Furthermore, 1% Ca-supplemented diet increased the levels of phosphorylated extracellular signal-regulated kinase and c-Jun N-terminal kinase. The increased expression of osteoblast differentiation marker genes and activation of mitogen-activated protein kinase signaling were associated with significant increases in trabecular bone volume, which plays an important role in the overall skeletal strength. Our results demonstrated that 1% Ca supplement inhibited osteoclastogenesis, stimulated osteoblastogenesis and restored bone loss in OVX rats.

Hederagenin Supplementation Alleviates the Pro-Inflammatory and Apoptotic Response to Alcohol in Rats.

In this study, we determined the effects of hederagenin isolated from Akebia quinata fruit on alcohol-induced hepatotoxicity in rats. Specifically, we investigated the hepatoprotective, anti-inflammatory, and anti-apoptotic effects of hederagenin, as well as the role of AKT and mitogen-activated protein kinase (MAPK) signaling pathways in ethanol-induced liver injury. Experimental animals were randomly divided into three groups: normal (sham), 25% ethanol, and 25% ethanol + hederagenin (50 mg/kg/day). Each group was orally administered the respective treatments once per day for 21 days. Acetaldehyde dehydrogenase-2 mRNA expression was higher and alcohol dehydrogenase mRNA expression was lower in the ethanol + hederagenin group than those in the ethanol group. Pro-inflammatory cytokines, including TNF-α, IL-6, and cyclooxygenase-2, significantly increased in the ethanol group, but these increases were attenuated by hederagenin. Moreover, Western blot analysis showed increased expression of the apoptosis-associated protein, Bcl-2, and decreased expression of Bax and p53 after treatment with hederagenin. Hederagenin treatment attenuated ethanol-induced increases in activated p38 MAPK and increased the levels of phosphorylated AKT and ERK. Hederagenin alleviated ethanol-induced liver damage through anti-inflammatory and anti-apoptotic activities. These results suggest that hederagenin is a potential candidate for preventing alcoholic liver injury.

Predictive factors and pattern of central lymph node metastasis in unilateral papillary thyroid carcinoma.

OBJECTIVE: Prophylactic central neck dissection (CND) for papillary thyroid carcinoma (PTC) remains controversial. The aim of this study was to evaluate the patterns and predictive factors of central lymph node metastasis in cases of PTC that were clinically determined to be node negative. METHODS: We studied 485 patients who have unilateral PTC without clinical lymph node metastasis and underwent total thyroidectomy and prophylactic bilateral CND from 2003 to 2012, retrospectively. The frequency, subsite and predictive factors of central lymph node metastasis were analyzed. RESULTS: In total, 166 (32.4%) patients had occult central lymph node metastases. The most common subsite of central node metastases was the ipsilateral paratracheal lymph node (26.0%), followed by pretracheal (12.5%), prelaryngeal (5.0%), and contralateral paratracheal (3.9%) lymph nodes. The tumor size larger than 0.5cm (p=0.003), age under 45 (p<0.001) and extrathyroidal extension (p=0.028) were associated with ipsilateral central compartment metastasis in multivariate analysis. Contralateral central node metastasis was associated with tumor size >3cm, age under 45, and multifocality and ipsilateral central node metastasis in univariate analysis, but it was associated with only ipsilateral central node metastasis in multivariate analysis (p=0.001). CONCLUSION: Prophylactic CND might be considered for PTC patients with large tumor size or extrathyroidal extension based on rates of lymph node metastasis. Unilateral CND might be appropriate as prophylactic CND due to the low metastatic rate to the contralateral paratracheal node.

Nasal and exhaled nitric oxide in chronic rhinosinusitis with polyps.

BACKGROUND: The ciliary epithelial cells in the paranasal sinuses produce nasal nitric oxide (NO) continuously and plays a variety of roles in the paranasal sinuses. The purpose of this study was to assess whether we can use the levels of nasal NO (nNO) and exhaled NO (eNO) as a tool for evaluation in chronic rhinosinusitis (CRS) with nasal polyp patients. METHODS: We used chemiluminescent NO analyzer to measure nNO and eNO among normal controls (32) and CRS with polyp (30) and CRS with polyp and allergic rhinitis patients (27) and compared it with various clinical symptoms, laboratory data, and computed tomography (CT) scores. RESULTS: Levels of nNO were significantly lower in patients with CRS with polyps (88.5 ± 54.7 ppb) compared with controls (241.0 ± 89.5 ppb). Levels of nNO in CRS with polyps and allergic rhinitis (167.0 ± 47.6 ppb) were significantly higher than CRS with polyps and lower than controls. A significant inverse relationship was observed between nNO and sinus CT scores, severity of nasal obstruction, and purulent rhinorrhea in CRS with polyps. Low values of nNO separated well patients with CRS with polyps, and the cutoff value of <163 ppb was associated with the best combination of specificity (93%) and sensitivity (81%). A significant positive relationship was observed between eNO and CT scores. CONCLUSION: The nNO could be used for another screening of CRS with polyps for the more severe phenotypes, which may eventually have to be treated with surgery.