Bone histomorphometric measurements are required to understand the efficacy
of treatment on bone remodelling. Previous studies used the Weibel technique
as a quantitative stereological method to determine bone cellular and dynamic
changes. However, there was no description on how this technique was applied.
This studyaimed to provide a full picture about the utilization of the Weibel
technique to measure static and dynamic bone histomorphometric indices.
Technical expertise, processing of bone samples, randomization of the trabecular
sections and an adequate number of analysed images for each section are required to achieve reliable results with a low possibility of errors.
Phytosterols are plant sterols with a chemical structure similar to cholesterol. It has anti-cholesterol, anti-cancer and anti-oxidant properties which are probably mediated by suppression of lipid peroxidation. However, there are limited studies on the effects of phytosterols on lipid peroxidation. The aim of this study is to determine the effects of phytosterols on plasma and tissue malondialdehyde (MDA) of rats exposed to carbon tetrachloride. The rats were divided into four groups of normal control (NC), carbon tetrachloride (CCl4), phytosterol (P) and phytosterol+carbon tetrachloride (P+CCl4).
The P and P+CCl4 groups were pretreated with subcutaneous phytosterol at 140 mg/kg once weekly for 5 weeks while the NC and CCl4 groups only received olive oil (vehicle). A single oral dose of carbon tetrachloride was then given to rats in the CCl4and P+CCl4 groups to induce lipid peroxidation. After 24 hours, all the rats were sacrificed and the plasma and tissue MDA were measured. Our results showed carbon tetrachloride had caused significant elevations of the plasma and hepatic MDA of the CCl4 group compared to the NC group. Phytosterol pretreatment (P+CCl4 group) were able to prevent the MDA elevations. Phytosterols treatments in normal rats (P group) were found to reduce the hepatic MDA level. The conclusion of this study was that phytosterols are effective suppressor of plasma and hepatic lipid peroxidation. They have potential as supplements to further reduce lipid peroxidation in healthy individuals.
Oxidative stress has been associated with postmenopausal osteoporosis which pre-disposes to risk of fracture. Palm tocotrienol is a potent antioxidant and has the poten-tial to be used for treatment of post-menopausal osteoporosis. The aim of the study is to determine if palm tocotrienol supplementation could alleviate oxidative stress in ovariectomised rat model and improve its bone strength. The rats were di- vided into four groups: (i) sham-operated group (SHAM) (ii) ovariectomised-control group (OVX) (iii) ovariectomised and given 60mg/kg α-tocopherol by oral gavage (OVX + ATF) (iv) ovariectomised and given 60mg/kg palm tocotrienols by oral gavage (OVX + PTT). After eight weeks of treatment, blood samples were taken to measure oxida-tive status (MDA, SOD and GPX) while the femurs were biomechanically tested for strength and resistance to fracture. Ovariectomy was shown to induce oxidative stress as shown by the raised MDA levels and reduced GPX activity. Palm tocotrienols seemed to offer protection against the ovariectomy-induced oxidative stress as shown by the suppression of MDA levels and raised GPX and SOD activities in the OVX+PTT group. In comparison, α-tocopherol was only able to raise the SOD but not as high as palm tocotrienols. The biomechanical tests have shown that ovariectomy has not af-fected the bone strength significantly after eight weeks. Palm tocotrienols supplemen-tation for eight weeks was effective in preventing oxidative stress in a post-meno-pausal rat.
Introduction: Childhood malnutrition is common and severe among indigenous community. The Community Feed-ing Program was first launched in 2010 among 15 villages in Kemar indigenous settlement among children below six years old. The objective of this study was to improve the nutrition status of indigenous children in Kemar settlement, Hulu Perak. Methods: All the indigenous children aged below six received high calorie food, full cream milk and multivitamin with an average of 500kcal/day. Ready to Use Therapeutic Food (RUTF), specified for malnourished children, provides nutrition that accounts for one-half to two-thirds of a child’s daily needs. The feeding sessions was carried out once a day, five days a week and managed by a group of trained local volunteers and research assistants. Weight and height were measured monthly. Results: The coverage throughout 2013 to 2018, ranged from 90.3% to 100%. The percentage of children with normal body weight had increased from only 38.7 % in 2010 to 60.6% in 2018. From year 2013 to 2018, the percentage of stunted children had reduced from 77% to 72.5%, and severe stunting reduced from 35.2% in 2015 to 24.9% in 2018. Conclusion: The continuity of this program is essential to sustain normal nutritional status and hence the wellbeing of this group of children in the interior remote community.
Long term glucocorticoids administration induces oxidative stress which leads to alteration of bone structure and strength. Palm oil is rich in tocotrienol, an antioxidant. It can be used for the prevention of oxidative stress related diseases. The main objective of this study was to determine the mechanism of palm tocotrienol in maintaining the bone structure and strength in glucocorticoidinduced osteoporosis. Thirty two adult male Sprague-Dawley rats, aged 3 months, weighing 300-320 g rats were used in this study. Sixteen rats undergone adrenalectomy and were administered with 120µg/kg/day intramuscular injection of dexamethasone. Eight rats were supplemented with oral palm tocotrienol 60 mg/kg/day (Adrx+Dex+PTT) and the other eight rats were given oral vehicle palm olein 0.1 ml/kg/day (Adrx+Dex). Eight rats underwent sham procedure and were given vehicle palm olein 0.05 ml/kg/day by intramuscularly and oral 0.1 ml/kg/day (Sham). The rats were euthanized after two months of treatments. Eight rats were euthanized after acclimatic action without receiving any treatment (Baseline). The right femurs were used for bone biomechanical strength and histomorphometry analysis while the left for gene expression and oxidative stress enzymes activities. The results indicated that long-term glucocorticoid treatment significantly increased bone resorption marker, CTX (6060.7 ± 410 pg/ml) and decreased bone structure and strength. Osteoblast and osteoclast related genes expressions indicated an increase in bone turnover. Supplementation of palm tocotrienol had maintained serum resorption (2619.4 + 209 pg/ml) marker level and preserved bone structure and strength. Gene expression analysis showed decrease in bone resorption. The findings suggested that palm tocotrienol has potential benefits against glucocorticoid-induced osteoporosis by regulating osteoblast and osteoclast related gene expressions.
Vitamin E deficiency has been found to impair bone calcification. This study was done to determine the effects of vitamin E deficiency and supplementation on parathyroid hormone, i.e. the hormone involved in bone regulation. Female Sprague-Dawley rats were divided into 4 groups: 1) normal rat chow (RC), 2) vitamin E deficiency (VED), vitamin E deficient rats supplemented with 3) 60 mg/kg alpha-tocotrienol (ATT) and 4) 60 mg/kg (alpha-tocopherol (ATF). Treatment was carried out for 3 months. Vitamin E deficiency caused hypocalcaemia during the first month of the treatment period, increased the parathyroid hormone level in the second month and decreased the bone calcium content in the 4th lumbar bone at the end of the treatment. Vitamin E supplementation (ATT and ATF) failed to improve these conditions. The bone formation marker, osteocalcin, and the bone resorption marker, deoxypyridinoline did not change throughout the study period. In conclusion vitamin E deficiency impaired bone calcium homeostasis with subsequent secondary hyperparathyroidism and vertebral bone loss. Replacing the vitamin E with pure ATF or pure ATT alone failed to correct the changes seen.
Pharmacology teaching during preclinical years is important for medical students to make rational choices in choosing suitable treatment for patients in future. Therefore, the present study determined the adequacy and effectiveness of pharmacology teaching in the undergraduate medical program at the Universiti Kebangsaan Malaysia Medical Center (UKMMC). Suggestions for improvement of the curriculum were also identified. An online questionnaire on the perceptions of pharmacology teaching methodology was distributed to a total of 459 medical students in 4th and 5th year at UKMMC. The questionnaire covered demographics, perceptions about pharmacology teaching, the ideal teaching learning methodology for learning pharmacology, pharmacology topics which are useful for future clinical practice, the pharmacology topic which was most interesting and recommendations for improvement. The response rate was 46.4% and majority of the participants were females (65.7%). Most of the students agreed that interactive learning was more helpful than didactic lectures (88.0%). Seventy percent of the students reported that pharmacology lectures in the preclinical years were helpful during the clinical years. Percentage of students who agreed that pharmacology teaching in their preclinical was adequate for their clinical practice was 47.0%. There was no association between demographic variables (gender, race, year of study and medical family background) and interest in pharmacology (p>0.05). In conclusion, the pharmacology teaching during preclinical years was perceived to be effective and useful for students’ clinical practice. More pharmacology teaching sessions in clinical years was suggested which may improve adequacy of pharmacology teaching.
There is growing evidence that inflammation may be one of the causal factors of osteoporosis. Several cytokines such as IL-1, IL-6, RANKL, OPG, and M-CSF were implicated in the pathogenesis of osteoporosis. These cytokines are important determinants of osteoclast differentiation and its bone resorptive activity. Anticytokine therapy using cytokine antagonists such as IL-receptor antagonist and TNF-binding protein was able to suppress the activity of the respective cytokines and prevent bone loss. Several animal studies have shown that vitamin E in the forms of palm-derived tocotrienol and α-tocopherol may prevent osteoporosis in rat models by suppressing IL-1 and IL-6. Free radicals are known to activate transcription factor NFκB which leads to the production of bone resorbing cytokines. Vitamin E, a potent antioxidant, may be able to neutralise free radicals before they could activate NFκB, therefore suppressing cytokine production and osteoporosis. Vitamin E has also been shown to inhibit COX-2, the enzyme involved in inflammatory reactions. Of the two types of vitamin E studied, tocotrienol seemed to be better than tocopherol in terms of its ability to suppress bone-resorbing cytokines.
Vitamin E has been shown to affect bone metabolism. In this study we determined the effects of palm vitamin E and alpha-tocopherol on bone metabolism. Sprague-Dawley female rats fed with normal rat chow were divided into 4 groups and supplemented with either palm vitamin E 30 mg/kg rat weight, palm vitamin E 60 mg/kg rat weight or alpha-tocopherol 30 mg/kg rat weight. One group was not supplemented. Half of these rats were ovariectomised before supplementation was given for 10 months. As expected, bone mineral density of the ovariectomised rats fed on normal rat chow diet was lower compared to the intact rats. However, these changes were not seen in the supplemented group of rats. Both intact and ovariectomised rats supplemented with palm vitamin E 30 mg/kg rat weight had a lower bone calcium content in both femoral and vertebral bones whilst rats fed palm vitamin E 60 mg/kg rat weight or alpha-tocopherol 30 mg/kg rat weight were able to maintain bone calcium content. Alkaline phosphatase activity was elevated in ovariectomised rats supplemented with palm vitamin E 30 mg/kg rat weight and alpha-tocopherol 30 mg/kg rat weight compared to the intact rats. Alpha-tocopherol also reduced the activity of tartrate-resistant acid phosphatase post-ovariectomy. These findings indicate that both palm vitamin E and alpha-tocopherol maintained bone mineral density in ovariectomised rats but caused conflicting effects on bone calcium content. Further study is needed in order to determine the mechanisms involved.
Osteoporosis is characterized by skeletal degeneration with low bone mass and destruction of microarchitecture of bone tissue which is attributed to various factors including inflammation. Women are more likely to develop osteoporosis than men due to reduction in estrogen during menopause which leads to decline in bone-formation and increase in bone-resorption activity. Estrogen is able to suppress production of proinflammatory cytokines such as IL-1, IL-6, IL-7, and TNF-α. This is why these cytokines are elevated in postmenopausal women. Studies have shown that estrogen reduction is able to stimulate focal inflammation in bone. Labisia pumila (LP) which is known to exert phytoestrogenic effect can be used as an alternative to ERT which can produce positive effects on bone without causing side effects. LP contains antioxidant as well as exerting anti-inflammatory effect which can act as free radical scavenger, thus inhibiting TNF-α production and COX-2 expression which leads to decline in RANKL expression, resulting in reduction in osteoclast activity which consequently reduces bone loss. Hence, it is the phytoestrogenic, anti-inflammatory, and antioxidative properties that make LP an effective agent against osteoporosis.
Vitamin E is an antioxidant that may protect bone against oxidative stress-induced osteoporosis. This in vitro study was conducted to determine the protective effects of a-tocopherol and γ-tocotrienol on osteoblasts, the bone forming cells, against oxidative stress.
Vitamin E is found to reverse the effects of nicotine on bone and this study aimed to determine its mechanism. Male Sprague Dawley rats were divided into four groups and treated for 3 months: Group 1 was the control group (RC). Groups 2 (N), 3 (N+TT) and 4 (N+ATF) received nicotine 7 mg/kg throughout the treatment period. In addition, groups 3 and 4 received tocotrienol 60 mg/kg and alpha-tocopherol 60 mg/kg respectively during months 2 and 3. Parameters measured were serum osteoprotegerin (OPG), serum receptor activator of nuclear factor kappa B ligand (RANKL), femoral and lumbar bone calcium content and body weight. Nicotine did not affect OPG or RANKL levels but reduced bone calcium content suggesting the calcium loss is not due to increase osteoclastogenesis. OPG was increased in N+ATF while RANKL was slightly increased in N+TT. Both vitamin E supplements restored bone calcium loss induced by nicotine. Nicotine impaired weight gain in all treatment groups starting week 4 however, N+TT group was comparable to RC from week 6 onwards. Bone protective effects of ATF, but not TT, may be partly due to inhibition of osteoclastogenesis.
This study was conducted to determine the effectiveness of three forms of vitamin E supplements following nicotine treatment on bone histomorphometric parameters in an adult male rat model. Rats were divided into seven groups: baseline (B, killed without treatment), control (C, normal saline for 4 months), nicotine (N, nicotine for 2 months), nicotine cessation (NC), tocotrienol-enhanced fraction (TEF), gamma-tocotrienol (GTT), and alpha-tocopherol (ATF). Treatments for the NC, TEF, GTT, and ATF groups were performed in two phases. For the first 2 months they were given nicotine (7 mg/kg), and for the following 2 months nicotine administration was stopped and treatments with respective vitamin E preparations (60 mg/kg) were commenced except for the NC group, which was allowed to recover without treatment. Rats in the N and NC groups had lower trabecular bone volume, mineral appositional rate (MAR), and bone formation rate (BFR/BS) and higher single labeled surface and osteoclast surface compared to the C group. Vitamin E treatment reversed these nicotine effects. Both the TEF and GTT groups, but not the ATF group, had a significantly higher trabecular thickness but lower eroded surface (ES/BS) than the C group. The tocotrienol-treated groups had lower ES/BS than the ATF group. The GTT group showed a significantly higher MAR and BFR/BS than the TEF and ATF groups. In conclusion, nicotine induced significant bone loss, while vitamin E supplements not only reversed the effects but also stimulated bone formation significantly above baseline values. Tocotrienol was shown to be slightly superior compared to tocopherol. Thus, vitamin E, especially GTT, may have therapeutic potential to repair bone damage caused by chronic smoking.
Adequate pain relief is a requisite for a successful closed manipulative reduction (CMR) of fractures and dislocations. This prospective study was undertaken to assess the mode and adequacy of pain relief given to patients undergoing such procedures at Seremban Hospital from the 1st April to the 31st May 2001. All patients with fractures and dislocations scheduled to undergo CMR were included in this study. The type of sedative agents and analgesia administered were recorded. Demographic data and the type of fracture or dislocation of the selected patients were documented. A visual analogue scale (VAS) for pain perception was given to both to the patients and the medical personnel who performed the procedure. All data were collected manually before entered into computerized database for analysis. Of 72 patients included in this study, 47% were Malay, 26% Indian, 21% Chinese and 6% others. There was male predominance and the patients' age ranged between 9 to 79 years (average 27.4 years). Upper limb injuries (79%) were mainly fractures of the radius and ulna (29%) and isolated fracture radius (21%). For the lower limb injuries (21%), combined tibia and fibula fractures constituted 10% of the total cases followed by isolated tibia fractures (10%) and hip dislocation (1%). The most common pain relieving agents given during the CMR were intravenous pethidine alone (43%) followed by combination of intravenous pethidine and valium (36%), intramuscular pethidine (17%) and intramuscular tramal (4%). The Visual Analogue Score (VAS) for pain perception revealed that 61% of the patients had moderate pain while 21% had severe pain during the course of the procedures. Suboptimal pain relief administered during CMR should prompt positive actions to ensure that the patient is not subjected to undue pain just for the sake of an acceptable fracture reduction.
In this study the effects of vitamin E deficiency and supplementation on bone calcification were determined using 4-month-old female Sprague-Dawley rats. The rats weighed between 180 and 200 g. The study was divided in three parts. In experiment I the rats were given normal rat chow (RC, control group), a vitamin E deficient (VED) diet or a 50% vitamin E deficient (50%VED) diet. In experiment 2 the rats were given VED supplemented with 30 mg/kg palm vitamin E (PVE30), 60 mg/kg palm vitamin E (PVE60) or 30 mg/kg pure alpha-tocopherol (ATF). In experiment 3 the rats were fed RC and given the same supplements as in experiment 2. The treatment lasted 8 months. Vitamin E derived from palm oil contained a mixture of ATF and tocotrienols. Rats on the VED and 50%VED diets had lower bone calcium content in the left femur compared to the RC group (91.6 +/- 13.3 mg and 118.3 +/- 26.0 mg cf 165.7 +/- 15.2 mg; P < 0.05) and L5 vertebra (28.3 +/- 4.0 mg and 39.5 +/- 6.2 mg compared with 51.4 +/- 5.8 mg; P < 0.05). Supplementing the VED group with PVE60 improved bone calcification in the left femur (133.6 +/- 5.0 mg compared with 91.6 +/- 13.3 mg; P < 0.05) and L5 vertebra (41.3 +/- 3.3 mg compared with 28.3 +/- 4.0 mg; P < 0.05) while supplementation with PVE30 improved bone calcium content in the L5 vertebra (35.6 +/- 3.1 mg compared with 28.3 +/- 4.0 mg; P < 0.05). However, supplementation with ATF did not change the lumbar and femoral bone calcium content compared to the VED group. Supplementing the RC group with PVE30, PVE60 or ATF did not cause any significant changes in bone calcium content. In conclusion, vitamin E deficiency impaired bone calcification. Supplementation with the higher dose of palm vitamin E improved bone calcium content, but supplementation with pure ATF alone did not. This effect may be attributed to the tocotrienol content of palm vitamin E. Therefore, tocotrienols play an important role in bone calcification.
The use of repeatedly heated frying oils and intake of high cholesterol diet have been linked to bone damage. The aim of this study is to determine the combined effects of taking repeatedly heated frying oils (palm or soy oil) and high cholesterol diet on the dynamic histomorphometric parameters of bone. Ovariectomised rats were used as animal model of post-menopausal osteoporosis. After six months of treatment, Double-labeled Surface (dLS/BS), Mineralising surface (MS/BS) and Bone Formation Rate (BFR/BS) of ovariectomised rats (OvxC) were significantly reduced compared to the normal control group. Additions of fresh or once-heated palm or soy oil into high cholesterol diet seem to have improved the dynamic parameters towards the normal control values. However, when these oils were repeatedly heated, the protective effects were lost and the dynamic parameters except MS/BS dropped back towards the ovariectomised-control values.
In this work, the effect of solvent pre-treatment (hexane, petroleum ether and ethanol) on the physicochemical, thermal and morphology behavior of Mangifera pajang seed fat (MPSF) were investigated. Fat extraction was performed using Soxhlet method, and results showed that the yield, physicochemical, and crystalline structures of the MPSF were significantly (p < 0.05) influenced by the extraction solvents. Hexane gave the highest fat yield (7.67 %) with low unsaturation value (52.13 g iodine/g) compared with petroleum ether and ethanol. Hexane MPSF also had low oxidation rate (peroxide value of 1.1 mEq/g). Both non-stabilized and stabilized hexane MPSF showed a single melting endothermic peak at high temperature with onset, maximum peak and offset temperature of 16.23 ˚C-18.21 °C, 28.22 ˚C-31.25 °C and 34.85 ˚C-39.58 °C, respectively. Hexane MPSF crystallized rapidly at high temperature with single maximum peak starting at 16.51 ˚C-16.68 °C and ending at 0.23 ˚C-1.13 °C. In comparison with ethanol extract, hexane MPSF demonstrated a compact crystalline structure with a large densely packed center. Therefore, MPSF obtained from hexane presented better overall quality than those obtained from other extraction solvents. MPSF exhibited similar melting and morphological behavior to mango kernel fat and commercial cocoa butter. These results suggested that hexane was the best solvent for the extraction of MPSF. This fat also has the potential to be applied as a cocoa butter alternative fat or functional fat.
Osteoporosis is a proven complication of long-term glucocorticoid therapy. Concern on glucocorticoid induced osteoporosis has increased dramatically in recent years with the widespread use of synthetic glucocorticoids. Glucocorticoid action in bone depends upon the activity of 11βhydroxysteroid dehydrogenase type 1 enzyme (11βHSD1). This enzyme plays an important role in regulating corticosteroids by locally interconverting cortisone into active cortisol. This has been demonstrated in primary cultures of human, mouse or rat osteoblasts. Therefore, inhibition of this enzyme may reduce bone resorption markers. Piper sarmentosum (Ps) is a potent inhibitor of 11βHSD1 in liver and adipose tissue. In this study we determined the effect of Ps on 11βHSD1 activity in bones of glucocorticoid-induced osteoporotic rats.