The impact of burial period on compact bone microstructure: Histological analysis of matrix loss and cell integrity in human bones exhumed from tropical soil.

Human bone histological analysis is a useful tool to assess post mortem diagenesis and to predict successful nuclear DNA typing of forensic material. This study is part of a series of studies developed by the authors intended to improve the understanding of post mortem diagenesis and to develop applications for DNA analysis of skeletal species from tropical soils, in order to optimize genetic and anthropological protocols. The aim of this study was to analyze the impact of burial period on the integrity of exhumed compact bone microstructure from tropical climate. In fragments of exhumed human femora from 39 individuals from the same cemetery (exhumed group) and 5 fresh femora from routine autopsies (control group), sections stained by hematoxylin-eosin were analyzed in order to measure bone microstructural integrity. We found that bone integrity index in exhumed group was negatively influenced by the period of burial (r = -0.37, p < 0.05) and highly significantly decreased (p < 0.0001) in comparison to control group. The period of burial and nitric acid decalcification time was positively correlated (r = 0.51; p < 0.01), leading to imply a bone petrification process during inhumation. Exhumed group showed higher level of matrix bone loss (p < 0.001), as expected, and 87% of cases analyzed were "tunneled" as described by Hackett. Bone integrity index and bone matrix tend to decrease in bones buried in tropical soil between 8-14 years of inhumation. This period is short if we consider cases in which there are preserved bones interred for longer periods in other environments. These data must be considered in cases where genetic identification of exhumed skeletons from tropical environment is required. The diagenesis in these bones and the variations of results found are discussed, clarifying some challenges for forensic laboratories, especially in DNA analysis.

Adhesives for fixed orthodontic brackets.

BACKGROUND: Bonding of orthodontic brackets to teeth is important to enable effective and efficient treatment with fixed appliances. The problem is bracket failure during treatment which increases operator chairside time and lengthens treatment time. A prolonged treatment is likely to increase the oral health risks of orthodontic treatment with fixed appliances one of which is irreversible enamel decalcification. This is an update of the Cochrane Review first published in 2003. A new full search was conducted on 26 September 2017 but no new studies were identified. We have only updated the search methods section in this new version. The conclusions of this Cochrane Review remain the same. OBJECTIVES: To evaluate the effects of different orthodontic adhesives for bonding. SEARCH METHODS: Cochrane Oral Health's Information Specialist searched the following databases: Cochrane Oral Health's Trials Register (to 26 September 2017), the Cochrane Central Register of Controlled Trials (CENTRAL; 2017, Issue 8) in the Cochrane Library (searched 26 September 2017), MEDLINE Ovid (1946 to 26 September 2017), and Embase Ovid (1980 to 26 September 2017). The US National Institutes of Health Ongoing Trials Register (ClinicalTrials.gov) and the World Health Organization International Clinical Trials Registry Platform were searched for ongoing trials. No restrictions were placed on the language or date of publication when searching the electronic databases. SELECTION CRITERIA: Trials were selected if they met the following criteria: randomised controlled trials (RCTs) and controlled clinical trials (CCTs) comparing two different adhesive groups. Participants were patients with fixed orthodontic appliances. The interventions were adhesives that bonded stainless steel brackets to all teeth except the molars. The primary outcome was debond or bracket failure. DATA COLLECTION AND ANALYSIS: Data were recorded on decalcification as a secondary outcome, if present. Information regarding methods, participants, interventions, outcome measures and results were extracted in duplicate by pairs of review authors. Since the data were not presented in a form that was amenable to meta-analysis, the results of the review are presented in narrative form only. MAIN RESULTS: Three trials satisfied the inclusion criteria. A chemical cured composite was compared with a light cured composite (one trial), a conventional glass ionomer cement (one trial) and a polyacid-modified resin composite (compomer) (one trial). The quality of the trial reports was generally poor. AUTHORS' CONCLUSIONS: There is no clear evidence on which to make a clinical decision of the type of orthodontic adhesive to use.

[MICROECOLOGICAL CHANGES IN THE INTESTINE AND ITS EFFECT ON THE CONDITION OF HARD TISSUES OF TEETH IN ADOLESCENCE].

On the background of microecological changes in the gastrointestinal tract, mineral and protein metabolism is disrupted, biochemical changes occur in the phosphorus-calcium metabolism in the bones of the skeleton and hard tissues of the teeth. The aim of the research was to study the activity of the caries process, characterized by progressive decalcification, under conditions of microecological changes in the intestine and its effect on the processes of endogenous calcium assimilation in adolescence. Within the frames of the research 68 adolescents aged 11 to 17 years with various degrees of pathology of the gastrointestinal tract were examined. The data of the performed studies showed that in patients with more severe changes in the intestinal micro flora, the prevalence and intensity of dental caries are significantly higher (83.4% on average DMFT 5.0) than in adolescents with mild dysbiosis (71.8% DMFT 3.2% ), and in the patients with normal intestinal micro flora - the parameters were minimal. Estimating and comparing the interrelation between the incidence of caries and the presence of diseases of the gastrointestinal tract in adolescents, a directly proportional dependence of the DMF index on age was found, but the greatest increase in the individual level of incidence of dental caries (SIC) was observed in adolescents aged 14-16 (SIC=0,42) with severe gastrointestinal pathology GIT. Thus, significant changes in dental status detected in adolescents with pathology of the gastrointestinal tract confirm the validity of complex treatment and preventive dental care, considering the processes occurring in the body and the oral cavity, as well as the systematic, purposeful dental care and the allocation of patients with gastrointestinal pathology (GIT) in a separate group.

Decalcification prevention around orthodontic brackets bonded to bleached enamel using different topical agents.

BACKGROUND: The present study was conducted to evaluate the effect of different topical agents utilized for prevention of enamel decalcification around orthodontic brackets bonded to bleached and non-bleached enamel. METHODS: Human maxillary premolars (n = 120) were divided into two equal groups. Teeth in group I were left without bleaching while those in group II were bleached with Vivastyle gel. Metal brackets were bonded to all the teeth using light-cured adhesive. Each group was divided into six equal subgroups (A, B, C, D, E, and F). In subgroup A, no material was applied (control). In subgroups B, C, D, E, and F, the following materials were applied respectively: Profluorid varnish, Enamel Pro Varnish, Ortho-Choice Ortho-Coat, GC Tooth Mousse, and GC MI Paste Plus. All teeth were cycled in a demineralization solution/artificial saliva for 15 days. Laser fluorescence was used to measure the level of enamel mineralization. The data were statistically analyzed. RESULTS: Regarding the non-bleaching subgroups, all studied material revealed significant demineralization reduction in comparison to the control subgroup (P < 0.05). Ortho-Choice Ortho-Coat revealed the highest significant effect while GC Tooth Mousse showed the least effect. In bleached subgroups, Profluorid varnish, Enamel Pro Varnish, and Ortho-Choice Ortho-Coat significantly reduced demineralization (P < 0.05) while either GC MI Paste Plus or GC Tooth Mousse had no significant effects (P > 0.05). CONCLUSIONS: Ortho-Choice Ortho-Coat, and Profluorid and Enamel Pro varnishes could be utilized successfully to reduce enamel demineralization around brackets bonded to either bleached or non-bleached enamel. GC MI Paste Plus and GC Tooth Mousse were effective only in non-bleached enamel.

Calcium orthophosphates (CaPO4): Occurrence and properties.

The present overview is intended to point the readers' attention to the important subject of calcium orthophosphates (CaPO4). This type of materials is of the special significance for the human beings because they represent the inorganic part of major normal (bones, teeth and antlers) and pathological (those appearing due to various diseases) calcified tissues of mammals. For example, atherosclerosis results in blood vessel blockage caused by a solid composite of cholesterol with CaPO4, while dental caries (tooth decay) and osteoporosis (a low bone mass with microarchitectural changes) mean a partial decalcification of teeth and bones, respectively, that results in replacement of a less soluble and harder biological apatite by more soluble and softer calcium hydrogenorthophosphates. Due to the compositional similarities to the calcified tissues of mammals, CaPO4 are widely used as biomaterials for bone grafting purposes. In addition, CaPO4 have many other applications. Thus, there is a great significance of CaPO4 for the humankind and, in this paper, an overview on the current knowledge on this subject is provided.

Risk of Malignancy in Thyroid Nodules with Non-Diagnostic Fine-Needle Aspiration: A Retrospective Cohort Study.

BACKGROUND: Fine-needle aspiration (FNA) of thyroid nodules is commonly performed, and despite the use of ultrasound (US) guidance, the rate of non-diagnostic FNAs is still significant. The risk of malignancy of thyroid nodules with a non-diagnostic FNA is not clearly defined. However, most studies exclude the majority of patients without a repeat biopsy or surgery, thus increasing the likelihood of selection bias. The aims of this study were to determine the malignancy risk in nodules with an initial non-diagnostic FNA, and to identify the factors associated with malignancy. METHODS: This was a retrospective cohort study of patients with thyroid nodules who underwent US-guided FNA between 2004 and 2010 and had a non-diagnostic result. Patients were followed until confirmatory diagnosis of the nature of the nodule was made. The outcome of malignant or benign disease was based on one of the following: (i) final surgical pathology following thyroidectomy; (ii) repeat biopsy; (iii) clinically, based on repeat ultrasound performed at least three years following biopsy; or (iv) report of thyroid status for patients without follow-up visits contacted by mail. RESULTS: There were 699 nodules from 665 patients included. The mean age was 59 ± 15 years, and 71.7% were women. There was complete follow-up of 495 nodules. After a median follow-up of 2.7 years, thyroid cancer was found in 15 nodules. The prevalence of malignancy was 3% (15/495). The presence of nodular calcifications was the strongest predictor of thyroid malignancy (odds ratio 5.03 [confidence interval 1.8-14.7]). Initial nodule size was inversely associated with malignancy (odds ratio 0.55 [confidence interval 0.28-0.93]). However, the 193 patients without follow-up had smaller nodules compared with those included in the analysis. None of the patients with repeatedly non-diagnostic results were diagnosed with thyroid cancer at follow-up. CONCLUSION: The prevalence of thyroid cancer in nodules with non-diagnostic results is lower than the malignancy rate in thyroid nodules in general, but not negligible. They should be followed as per guidelines with heightened suspicion for nodules containing calcifications. Nodules with repeatedly non-diagnostic FNA results especially in the absence of calcifications have a low risk of malignancy and may be observed.

Direct inhibition of osteoblastic Wnt pathway by fibroblast growth factor 23 contributes to bone loss in chronic kidney disease.

Bone loss and increased fractures are common complications in chronic kidney disease. Because Wnt pathway activation is essential for normal bone mineralization, we assessed whether Wnt inhibition contributes to high-phosphorus-induced mineralization defects in uremic rats. By week 20 after 7/8 nephrectomy, rats fed a high-phosphorus diet had the expected high serum creatinine, phosphorus, parathyroid hormone, and fibroblast growth factor 23 (FGF23) levels and low serum calcium. There was a 15% reduction in tibial mineral density and a doubling of bone cortical porosity compared to uremic rats fed a normal-phosphorus diet. The decreases in tibial mineral density were preceded by time-dependent increments in gene expression of bone formation (Osteocalcin and Runx2) and resorption (Cathepsin K) markers, which paralleled elevations in gene expression of the Wnt inhibitors Sfrp1 and Dkk1 in bone. Similar elevations of Wnt inhibitors plus an increased phospho-ß-catenin/ß-catenin ratio occurred upon exposure of the osteoblast cell line UMR106-01 either to uremic serum or to the combination of parathyroid hormone, FGF23, and soluble Klotho, at levels present in uremic serum. Strikingly, while osteoblast exposure to parathyroid hormone suppressed the expression of Wnt inhibitors, FGF23 directly inhibited the osteoblastic Wnt pathway through a soluble Klotho/MAPK-mediated process that required Dkk1 induction. Thus, the induction of Dkk1 by FGF23/soluble Klotho in osteoblasts inactivates Wnt/ß-catenin signaling. This provides a novel autocrine/paracrine mechanism for the adverse impact of high FGF23 levels on bone in chronic kidney disease.

Pediatric thyroid nodules: ultrasonographic characteristics and inter-observer variability in prediction of malignancy.

BACKGROUND: Pediatric thyroid nodules, while uncommon, have high malignancy risk. The objectives of the study were (1) to identify sonographic features predictive of malignancy; (2) to create a prediction model; and (3) to assess inter-observer agreement among radiologists. METHODS: All available cases of thyroid nodules, surgically removed between 2000 and 2009. Three radiologists reviewed the sonographic images; 2 pathologists reviewed the tissue specimens. Adult prediction models were applied. Interobserver variability was assessed. RESULTS: Twenty-seven subjects, mean age 13.1±3.4 years, were included. Nineteen nodules were differentiated thyroid carcinomas. On multivariate analysis, size was the only significant predictor of malignancy. On recursive partitioning analysis, size >35 mm with microcalcification and ill-defined margins yielded the best prediction model. Radiologist inter-observer agreement regarding malignancy was moderate (κ=0.50). CONCLUSIONS: Larger size, microcalcifications and ill-defined margins on ultrasound demonstrate the best predictive model for malignancy in the pediatric population. Experienced pediatric radiologists demonstrate moderate inter-observer agreement in prediction of malignancy.

Loss of c-Kit and bone marrow failure upon conditional removal of the GATA-2 C-terminal zinc finger domain in adult mice.

Heterozygous mutations in the transcriptional regulator GATA-2 associate with multilineage immunodeficiency, myelodysplastic syndrome (MDS), and acute myeloid leukemia (AML). The majority of these mutations localize in the zinc finger (ZnF) domains, which mediate GATA-2 DNA binding. Deregulated hematopoiesis with GATA-2 mutation frequently develops in adulthood, yet GATA-2 function in the bone marrow remains unresolved. To investigate this, we conditionally deleted the GATA-2 C-terminal ZnF (C-ZnF) coding sequences in adult mice. Upon Gata2 C-ZnF deletion, we observed rapid peripheral cytopenia, bone marrow failure, and decreased c-Kit expression on hematopoietic progenitors. Transplant studies indicated GATA-2 has a cell-autonomous role in bone marrow hematopoiesis. Moreover, myeloid lineage populations were particularly sensitive to Gata2 hemizygosity, while molecular assays indicated GATA-2 regulates c-Kit expression in multilineage progenitor cells. Enforced c-Kit expression in Gata2 C-ZnF-deficient hematopoietic progenitors enhanced myeloid colony activity, suggesting GATA-2 sustains myelopoiesis via a cell intrinsic role involving maintenance of c-Kit expression. Our results provide insight into mechanisms regulating hematopoiesis in bone marrow and may contribute to a better understanding of immunodeficiency and bone marrow failure associated with GATA-2 mutation.

Bilateral choroidal osteoma: visual acuity at 45-year follow-up.

PURPOSE: To report a case of bilateral choroidal osteoma, which demonstrated different visual outcomes in both eyes over a long follow-up of 45 years. METHODS: Case report. RESULTS: After 45 years of follow-up, choroidal osteoma showed slight enlargement in each eye and choroidal neovascularization and retinal pigment epithelium (RPE) changes had developed in the right eye. Visual acuity of the right eye was decreased to 20/200 from macular RPE loss and choroidal neovascularization scarring. The left eye preserved a visual acuity of 20/30 where choroidal osteoma demonstrated only partial decalcification of the nasal margin with preservation of RPE in the macular region. Spectral domain optical coherence tomography of the right macula confirmed broad RPE and choroidal atrophy and scarring related to previous choroidal neovascularization. Spectral domain optical coherence tomography of the left macula showed preserved retinal layers and RPE with underlying calcified choroidal osteoma. CONCLUSION: Choroidal osteoma can occasionally demonstrate intact calcification, preserved RPE, and photoreceptors with excellent vision.

[Association of consumption of carbonated beverages and decalcification in woman on reproductive and non-reproductive age of Mexico City]. / Asociación del consumo de bebidas carbonatadas y descalcificación en mujeres en edad reproductiva y no reproductiva de la Ciudad de México.

INTRODUCTION AND OBJECTIVE: There is not enough information about carbonated beverages (CB) consumption in Mexican women and its association with bone mineral density (BMD). The objective was to identify the association of CB consumption with BMD in two groups of Mexican women. METHODS: Transversal study with 328 women in reproductive age (WRA) and not reproductive (WNRA) with diet and anthropometric evaluation and BMD. RESULTS: Mean age of 18.7 years in WRA and 47 in WNRA. Energy adequacy percentage was lower in WNRA (98 ± 28% vs 144 ± 55) (p = 0.001), calcium intake was < 700 mg/day in women with osteopenia of both groups and median consumption of CB was higher in WRA, with 200 mL/day (0-462) vs 0 (0-250) in WNRA (p = 0.001), WRA with osteopenia drank 500 mL/day (253-750) of CB vs 100 (0-200) in who had not (p = 0.001). Anthropometric parameters were higher in WNRA (p =?0.002) and femoral neck BMD was lower (1.031 ± 0.14 vs 1.107 ± 0.10) (p = 0.001) and higher in L2-L4 (1.114 ± 0.13 vs 1.003 ± 0.09) (p = 0.001) in WRA. Variables associated with risk of osteopenia: drink CB (OR 11.186, p = 0.001), consuming < 700 mg of calcium (OR 5.774, p = 0.001) and dinner no milk (OR 1.942, p = 0.042). CONCLUSIONS: To drink CB increases risk of osteopenia in both groups, WRA drink more CB than WNRA and they will have high probability of fractures younger.

Introducción y objetivo: No hay información suficiente sobre el consumo de bebidas carbonatadas (BC) y su asociación con la densidad mineral ósea (DMO) en mexicanas. El objetivo fue identificar la asociación del consumo de BC con la DMO en dos grupos de mujeres mexicanas. Métodos: Estudio transversal con 328 mujeres en edad reproductiva (MER) y no reproductiva (MENR), con evaluación dietética, antropométrica y DMO. Resultados: Edad promedio de 18,7 años en MER y 47 años en MENR. El porcentaje de adecuación del consumo de energía fue inferior en MENR (98 ± 28% vs 144 ± 55) (p = 0,001), el consumo de calcio fue < 700 mg/día en mujeres con osteopenia de ambos grupos y la mediana del consumo de BC fue mayor en MER, con 200 mL/día (0- 462) vs 0 (0-250) en MENR (p = 0,001), las MER con osteo penia bebían 500 mL/día (253-750 mL/día) de BC/día vs 100 (0-200 mL/día) en las que no la presentaron (p = 0,001). Los parámetros antropométricos fueron superiores en MENR (p =?0.002) y la DMO en cuello de fémur fue inferior 1,031 ± 0,14 vs 1,107 ± 0,10 (p = 0,001), en MER fue mayor en L2-L4: 1,114 ± 0,13 vs 1,003 ± 0,09 (p = 0,001). Variables asociadas al riesgo de osteopenia: beber BC (OR 11,186; p = 0,001), consumir < 700 mg de calcio (OR 5,774; p = 0,001) y no cenar leche (OR 1,942; p = 0,042). Conclusiones: Beber BC aumenta el riesgo de osteopenia en ambos grupos, las MER consumen más BC que las MENR y tienen aún más probabilidad de fracturas a edades más tempranas.

Importance of citrate and the calcium : citrate ratio in patients with calcium renal lithiasis and severe lithogenesis.

UNLABELLED: Different studies have shown the importance of citrate in the formation of calcium stones. It has further been shown that the states of metabolic acidosis result in an increase in bone resorption and lower urinary citrate levels. Increasing the intake of citrate in these patients can reduce the lithogenic risk and improve bone mineral density (BMD), contributing to control of both diseases. The study shows the importance of citrate in patients with calcium stones and BMD loss. The deficit in citrate excretion is associated with a decrease in bone mineralization and increased ß-crosslaps. A calcium : citrate ratio >0.25 in patients with calcium stones and loss of mineral density may predict severe lithogenic activity. OBJECTIVE: To analyse the importance of urinary citrate and the urinary calcium : citrate ratio in patients with calcium renal lithiasis and severe lithogenesis compared with a control group of patients without lithiasis. MATERIAL AND METHODS: A cross-sectional study of 115 patients in eastern Andalusia, Spain was conducted. The patients were divided into two groups: Group A: 56 patients aged 25-60 years without calcium renal lithiasis; Group B: 59 patients aged 25-60 years, presenting with calcium renal lithiasis and severe lithogenesis. The citrate levels and the calcium : citrate ratio in the patients' urine and the relationship of these two factors to lithiasic activity were analysed and compared. RESULTS: In Group B, 32.2% of the patients presented with hypocitraturia, compared with 14.3% of the patients in Group A (P = 0.02). The urinary citrate levels were lower in Group B than in Group A (P = 0.001) and the calcium : citrate ratio was higher in Group B than in Group A (P = 0.005). The results suggest that a patient urinary calcium : citrate ratio > 0.25 indicates severe lithogenesis (with a sensitivity of 89% and a specificity of 57%). After linear regression analysis, we found that the urinary citrate level is an independent factor associated with the changes in bone densitometry T-score values of patients. CONCLUSIONS: The patients with severe lithogenesis presented with hypocitraturia, which was associated with lower bone mineral density. The calcium : citrate ratio, which is linearly related to the bone resorption marker ß-crosslaps, could be useful in evaluating the risk of severe lithogenesis when this ratio is >0.25.

Space flight calcium: implications for astronaut health, spacecraft operations, and Earth.

The space flight environment is known to induce bone loss and, subsequently, calcium loss. The longer the mission, generally the more bone and calcium are lost. This review provides a history of bone and calcium studies related to space flight and highlights issues related to calcium excretion that the space program must consider so that urine can be recycled. It also discusses a novel technique using natural stable isotopes of calcium that will be helpful in the future to determine calcium and bone balance during space flight.

Multimodal fundus imaging in choroidal osteoma.

PURPOSE: To describe the morphologic features of calcified and decalcified choroidal osteomas using multimodal imaging and correlate these findings with a previous histopathologic study. DESIGN: Retrospective observational case series. METHODS: Three patients with choroidal osteoma underwent complete ophthalmologic examination, fundus photography, and multimodal fundus imaging, including Fourier-domain optical coherence tomography (FD-OCT) and blue-light fundus autofluorescence (bAF). RESULTS: FD-OCT imaging of calcified tumors revealed a distinctive latticework pattern of reflectivity resembling the spongy bone structure seen histopathologically. On bAF the fluorescence was relatively well preserved overlying calcified tumors. In decalcified areas 2 patterns of reflectivity were identified: the first consisted of areas of relative hyperreflectivity with a lamellar appearance while the second was characterized by heterogeneous, hyperreflective, mound-like irregular areas associated with some posterior optical shadowing. Decalcified tumor areas had reduced overall fluorescence on bAF. CONCLUSION: FD-OCT demonstrated different reflectivity patterns in both calcified and decalcified portions of the choroidal osteoma, which may correspond to different stages of tumor evolution. A distinctive latticework pattern of reflectivity similar to spongy bone was seen in calcified tumors. These observations improve our knowledge of the in vivo structure of choroidal osteomas and may have implications for the diagnosis and management of this tumor.

Spectral-domain optical coherence tomography of the choroid in choroidal osteoma.

The authors report spectral-domain optical coherence tomography (SD-OCT) findings in a patient with decalcifying choroidal osteoma accompanied by a choroidal neovascular membrane and serous retinal detachment. A 13-year-old girl was found to have a choroidal osteoma in the left eye. The clinical diagnosis was confirmed by B-scan ultrasonography, computed tomography, and fluorescein and indocyanine green angiography. The SD-OCT findings over the decalcified portion included serous retinal detachment, photoreceptor outer segment disorganization, retinal pigment epithelial atrophy, deformed Bruch's membrane, and choroidal neovascular membrane. In contrast, the retinal structures over the calcified portion appeared to be preserved. SD-OCT showed loss of a vascular appearance and increased thickness in the affected choroid, especially in the decalcified portion. Choroidal thickening may be associated not only with choroidal osteomas, but also with tumor decalcification. These unique features on SD-OCT may be important in understanding poor visual prognosis when decalcification involves the fovea.

The anti-osteoporotic effect of Eurycoma Longifolia in aged orchidectomised rat model.

Osteoporosis in elderly men is becoming an important health issue with the aging society. Elderly men with androgen deficiency are exposed to osteoporosis and can be treated with testosterone replacement. In this study, Eurycoma longifolia (EL), a plant with androgenic effects, was supplemented to an androgen-deficient osteoporotic aged rat as alternative to testosterone. Aged 12 months old Sprague-Dawley rats were divided into groups of normal control (NC), sham-operated (SO), orchidectomised-control (OrxC), orchidectomised and supplemented with EL (Orx + El) and orchidectomised and given testosterone (Orx + T). After 6 weeks of treatment, serum osteocalcin, serum terminal C-telopeptide Type 1 collagen (CTX) and the fourth lumbar bone calcium were measured. There were no significant differences in the osteocalcin levels before and after treatment in all the groups. The CTX levels were also similar for all the groups before treatment. However, after treatment, orchidectomy had caused significant elevation of CTX compared to normal control rats. Testosterone replacements in orchidectomised rats were able to prevent the rise of CTX. Orchidectomy had also reduced the bone calcium level compared to normal control rats. Both testosterone replacement and EL supplementation to orchidectomised rats were able to maintain the bone calcium level, with the former showing better effects. As a conclusion, EL prevented bone calcium loss in orchidectomised rats and therefore has the potential to be used as an alternative treatment for androgen deficient osteoporosis.