Osteogenic Induction with Silicon Hydroxyapatite Using Modified Autologous Adipose Tissue-Derived Stromal Vascular Fraction: In Vitro and Qualitative Histomorphometric Analysis.

Large bone defects requiring invasive surgical procedures have long been a problem for orthopedic surgeons. Despite the use of autologous bone grafting, satisfactory results are often not achieved due to associated limitations. Biomaterials are viable alternatives and have lately been used in association with Stromal Vascular Fraction (SVF), stem cells, and signaling factors for bone tissue engineering (BTE). The objective of the current study was to assess the biocompatibility of Silicon Hydroxyapatite (Si-HA) and to improve osteogenic potential by using autologous adipose-derived SVF with Si-HA in a rabbit bone defect model. Si-HA granules synthesized using a wet precipitation method were used. They were characterized using scanning electron microscopy (SEM), Fourier transform infrared (FTIR), and X-ray diffraction (XRD). A hemolysis assay was used to assess the hemolytic effects of Si-HA, while cell viability was assessed through Alamar Blue assay using MC3T3 mouse osteoblasts. The osteogenic potential of Si-HA both alone and with enzymatically/non-enzymatically-derived SVF (modified) was performed by implantation in a rabbit tibia model followed by histomorphometric analysis and SEM of dissected bone after six weeks. The results showed that Si-HA granules were microporous and phase pure and that the addition of Silicon did not influence Si-HA phase composition. Si-HA granules were found to be non-hemolytic on the hemolysis assay and non-toxic to MC3T3 mouse osteoblasts on the Alamar Blue assay. Six weeks following implantation Si-HA showed high biocompatibility, with increased bone formation in all groups compared to control. Histologically more mature bone was formed in the Si-HA implanted along with non-enzymatically-derived modified SVF. Bone formation was observed on and around Si-HA, reflecting osseointegration. In conclusion, Si-HA is osteoconductive and promotes osteogenesis, and its use with SVF enhances osteogenesis.

Adverse Effects of Caffeine on Development of Femur of BALB/c Mice and Protective Role of Vitamin D3.

OBJECTIVE: To determine the effects of caffeine ingestion on the development of femur and role of vitamin D3 in preventing these effects in BALB/c mice. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Department of Anatomy, Army Medical College, Rawalpindi, in cooperation with NIH (National Institute of Health), Islamabad, from October 2014 to October 2015. METHODOLOGY: Thirty (100%) BALB/c mice, 50% male and female each, three weeks old, weighing 12-14 grams were taken and divided equally and randomly into three groups, each having 10 (33.3%) mice; 5 (16.6%) male and female. G1 (control group) was given normal diet with water ad libitum. G2and G3(experimental groups) were given 10 mg of caffeine per 100g body weight, three days a week, through oral gavage for 60 days on alternate days. However, experimental group G3was additionally provided 0.1µg vitamin D3daily, through oral gavage for 60 days. Experimental groups were compared with control group and data was analyzed statistically. RESULTS: The mean weight of mice femur of G1(control group) was 0.387 ±0.019 g; while mean weights of right femur of G2and G3(experimental groups) were 0.316 ±0.020 g and 0.345 ±0.020 g, respectively. Similarly, mean right femur length of group G1 was 20.70 ±0.609 mm; while for groups G2 and G3, it was 24.382 ±1.087 mm and 22.966 ±0.822 mm, respectively. In comparison with group G1for groups G2and G3, femur weight decreased, however femur length increased. CONCLUSION: Caffeine intake caused femur length to increase and weight to decrease, but treatment with vitamin D3.

Histological changes induced by tamoxifen versus tamoxifen plus 13-cis-retinoic acid on rabbit uterine glands.

OBJECTIVE: To study the effects of tamoxifen versus tamoxifen plus 13-cis-retinoic acid on the histology of uterine glands in rabbits. METHODS: The experimental, randomised, controlled trial was conducted at the Army Medical College, Rawalpindi, from March 2009 to June 2009 and comprised rabbits acquired from the National Institute of Health, Islamabad. The animals were randomly divided into three equal groups: group A had controls, group B was treated with tamoxifen, and group C with tamoxifen plus retinoic acid. The uterine weight and cross-sectional diameter of uterine horns were measured after sacrifice. The uteri were processed for paraffin embedding. The sections were then assessed for stratification of glandular epithelium, changes in the glandular shape and glandular epithelial height. SPSS 13 was used for statistical analysis. RESULTS: Tamoxifen administration resulted in variation of glandular shape and increase in glandular epithelial height in group B as compared to control group, p < 0.001 and 0.005 respectively. The adjuvant administration of 13-cis-retinoic acid showed a suppressive effect only on glandular epithelial height, when compared with Group B (p=0.01). CONCLUSIONS: The 13-cis-retinoic acid has no significant inhibitory effect on uterine glandular proliferation induced by tamoxifen after a short-term administration of three months.

Amelioration of lead induced changes in ovary of mice, by garlic extract.

OBJECTIVE: To observe the effects of lead acetate and protective role of garlic extract on the histomorphology of the ovary in an animal model. METHODS: The experimental trial was conducted at the Department of Anatomy, Army Medical College Rawalpindi, in association with the National Institute of Health (NIH), Islamabad, from April to June 2013. It comprised 30 adult non-pregnant female mice (BALBc strain) weighing 25-27 gms. They were divided into three equal groups of 10 mice each. Group A, taken as control, was given normal diet. Group B was given lead acetate at a dose of 30 mg/kg/day. Group C was given lead acetate 30 mg/kg/day and garlic extract 500 mg/kg/day through oral gavage tube for two months. Animals were dissected a day after the last dose. Size, shape, colour and consistency of the ovary was observed. The right ovary was processed, embedded and stained for histological study. Primary follicles were counted and noted. SPSS 18 wsa used for statistical analysis. RESULTS: The primary follicular count decreased significantly in Group B while it relatively increased in Group C. Morphology of the ovary was affected after exposure to lead acetate in Group B, while in Group C results were the same as in the Group A controls regarding gross architecture of the ovary. CONCLUSION: Lead alters the normal histology and affects the physiology of the ovary. It interferes with the development of growing follicles in the ovary. Lead, being a reproductive toxicant, can be a cause of infertility in exposed females.

Patterns of sudanophilic lesions at ostia of descending thoracic and abdominal aorta in cholesterol-fed adult rabbits / Patrones de lesiones sudanofílicas en el ostio de las partes torácica y abdominal de la aorta en conejos adultos alimentados con colesterol

Sudan-positive atherosclerotic lesions preferentially occur at downstream and lateral margins of arterial branch ostia in human neonates and weanling rabbits but tend to develop at lateral and upstream margins in old subjects. We investigated (i) the pattern of sudanophilic lesions at aortic ostia of cholesterol-fed adult rabbits and (ii) determined any differences in lesion distribution between descending thoracic and abdominal aorta. Ten adult males, New Zealand white rabbits were fed 2 percent high cholesterol diet. After six weeks, aortas were excised, opened longitudinally and stained with Sudan-IV for gross examination of atherosclerotic lesions. A total of 156 descending thoracic and 90 abdominal ostia were examined. Mean lesion frequencies upstream, downstream and at lateral margins of the affected ostia were calculated and compared. Sudanophilic lesions were detected around 32 percent ostia of descending thoracic aorta and 25 percent those of abdominal aorta. At ostia of descending thoracic aorta, lesion frequencies were significantly higher (P<0.001) downstream (95 percent) and at lateral margins (92 percent) than upstream (2 percent). In abdominal aorta, lateral (100 percent) and upstream (43 percent) margins were significantly (P<0.05) more affected while minimal lesion frequencies were seen at downstream branch points (9 percent). Comparison between descending thoracic and abdominal aorta showed an insignificant difference of lesion frequencies at lateral margins (P>0.05) but a highly significant difference at upstream versus downstream of ostia (P<0.001). We concluded that in cholesterol-fed adult rabbits, juvenile pattern of downstream lipid deposition persists at ostia of descending thoracic aorta while a switch towards the upstream pattern of old subjects occurs at ostia of abdominal aorta.

Las lesiones ateroscleróticas Sudán-positivas se producen preferentemente en los márgenes posteriores y laterales del ostio de las ramas arteriales en neonatos humanos y en conejos destetados, pero tienden a desarrollarse en los márgenes laterales y superiores en sujetos de edad avanzada. Investigamos el patrón de lesiones sudanofílicas en el ostio aórtico de conejos adultos alimentados con colesterol y determinamos las diferencias en la distribución de lesiones entre la aorta torácica descendente y abdominal. Diez conejos machos blancos adultos New Zealand fueron alimentados con una dieta alta en colesterol al 2 por ciento. Después de seis semanas, fueron extraídas las aortas, se disecaron longitudinalmente y se tiñeron con Sudan-IV para el examen macroscópico y se observaron las lesiones ateroscleróticas. Fueron examinados 156 ostios de aortas torácicas descendentes y 90 de aortas abdominales. Fueron calculadas y comparadas las frecuencias medias de lesiones superiores, inferiores y de los márgenes laterales de los ostios afectados. Las lesiones sudanofílicas se detectaron en alrededor del 32 por ciento de ostios de la aorta torácica descendente y en el 25 por ciento de las aortas abdominales. En el ostio de la aorta torácica descendente, las frecuencias de lesiones fueron significativamente mayores (p <0,001) por superior (95 por ciento) y en los márgenes laterales (92 por ciento) que por inferior (2 por ciento). En la aorta abdominal, los márgenes laterales (100 por ciento) y superiores (43 por ciento) fueron significativamente más afectados (p <0,05), mientras que las frecuencias mínimas de lesiones se observaron en los puntos de la rama descendente (9 por ciento). La comparación entre la aorta torácica descendente y abdominal mostró una diferencia no significativa de las frecuencias de lesiones en los márgenes laterales (p> 0,05), pero muy significativa al comparar el margen superior con el inferior (p <0,001). Llegamos a la conclusión que en los conej...