Gross anatomical studies on the hind limb of the Sitatunga (Tragelaphus spekii)

The Sitatunga, Tragelaphus spekii, is a swamp dwelling antelope resident in West Africa. This study was carried out to document unique morphological and numerical information on the hind limb bones of this ruminant. Two (2) adults of both sexes were obtained as carcass at different times after post-mortem examination and prepared to extract the bones via cold water maceration for use in the study. The presence of a sharp pointed ilio-pubic eminence at the junction between the cranial border of the ilium and pubis; less prominent ischial tuber, inconspicuous ischiatic arch and a large oval obturator foramen were unique features of the Ossa coxarum that distinguished it from that of small ruminants. The Femur’s medial condyle was obliquely orientated, the fibula was absent while the long Tibia was typical of ruminant presentation. It was observed that the morphological features of the tarsals and Pes were also typical. However, the last Phalanges presented characteristic long triangular shaped bones with sharp pointed ends. The total number of bones making up the forelimb was accounted to be 45. In conclusion, this study has provided a baseline data for further biological, archeological and comparative anatomical studies.&nbsp

Formulation and evaluation of ciprofloxacin-cockle shells derived calcium carbonate aragonite nanoparticles physicochemical properties mediated in vitro bactericidal activity in Salmonella Typhimurium

Since the use of classical antibiotics in the management of resistant bacterial infections requires high dosage and regular administration for a lengthy duration, the enhanced delivery system that will ensure sustained release of antibiotic to the Site of action is important. We synthesized and formulated ciprofloxacin-cockle shell derived calcium carbonate aragonite nanoparticles (C-CSCCAN), appropriately analysed its physicochemical properties mediated antibacterial activity in Salmonella Typhimurium. The size of the formulated nanoparticles were in the range of 13.94 and 23.95 nm and Zeta potential was optimally negative. Diffraction pattern by X-ray powder diffraction (XRD) revealed strong crystallizations in all the formulations. Fourier-transform (FT-IR) spectra displayed evidence of interactions between the drug and nanoparticles at the molecular level and no change in peaks position was observed prior to and after the synthesis of the nanoparticles. Higher encapsulation (99.5) and loading capacity (5.9%) were attained at ciprofloxacin to nanoparticles ratio 1:17. No burst effect but a sustained drug release was observed from the formulation. C-CSCCAN suspension exhibited higher antibacterial activity than free ciprofloxacin. It was concluded that physicochemical properties of CSCCAN enhanced susceptibility of Salmonella Typhimurium, which could potentially improve the clinical efficacy of ciprofloxacin

Sperm attributes and morphology on Rusa timorensis: light and scanning electron microscopy

This study provides standard information on the attributes of sperm and describes the surface structure of normal and abnormal spermatozoa of Rusa timorensis. Two fertile stags were used as the source of semen collected during the first breeding season commencing from April 5 to July 2, 2012. Another five stags were used as the source of semen collected during the second breeding season commencing from April 1 to June 27, 2013. Semen samples were collected from the stags using an electro-ejaculator. The ejaculate was processed and samples prepared for light and scanning electron microscopy (SEM) according to standard methods. No significant difference (P > 0.05) was found between sperm attributes in comparison between different stags and different months of the fertile seasons. The results of this study have also demonstrated that there are no differences in size, shape and surface structure between spermatozoa of the different stags and different months of the fertile seasons. Sperm attributes (volume, pH, sperm concentration, general motility, progressive motility and viability) were 2.2 ± 0.29 ml, 7.2 ± 0.17, 886.3 ± 39.7 × 106 spermatozoa/ml, 78.7 ± 2.01%, 80.8 ± 1.85% and 83.2 ± 0.85%, respectively. Morphological analysis showed low percentage of abnormal spermatozoa 13.9 ± 2.88%. Scanning electron microscopy revealed spermatozoa which consisted of a flat paddle-shaped head, short neck and a tail, which was subdivided into midpiece, principal piece and endpiece. The average spermatozoon was 66.2 ± 0.69 μm in total length. The flat paddle-shaped head was 7.8 ± 0.28 μm long, 4.2 ± 0.15 μm at its widest width, 2.4 ± 0.18 μm basal width and 0.7 ± 0.02 μm thick. As for the tail, the midpiece length was 13.2 ± 0.14 μm, 0.6 ± 0.04 μm in diameter; the principal piece was 42.6 ± 0.04 μm, and 2.8 ± 0.06 μm for the endpiece. Abnormal spermatozoa such as tapered head, microcephalic head, decapitated spermatozoa and bent tails were observed. Results provide standard information useful for development of strategies for semen cryopreservation and assisted reproductive technology in this species

Safety assessments of subcutaneous doses of aragonite calcium carbonate nanocrystals in rats

Calcium carbonate nanoparticles have shown promising potentials in the delivery of drugs and metabolites. There is however, a paucity of information on the safety of their intentional or accidental over exposures to biological systems and general health safety. To this end, this study aims at documenting information on the safety of subcutaneous doses of biogenic nanocrystals of aragonite polymorph of calcium carbonate derived from cockle shells (ANC) in Sprague-Dawley (SD) rats. ANC was synthesized using the top-down method, characterized using the transmission electron microscopy and field emission scanning electron microscope and its acute and repeated dose 28-day trial toxicities were evaluated in SD rats. The results showed that the homogenous 30 ± 5 nm-sized spherical pure aragonite nanocrystals were not associated with mortality in the rats. Severe clinical signs and gross and histopathological lesions, indicating organ toxicities, were recorded in the acute toxicity (29,500 mg/m2) group and the high dose (5900 mg/m2) group of the repeated dose 28-day trial. However, the medium- (590 mg/m2 body weight) and low (59 mg/m2)-dose groups showed moderate to mild lesions. The relatively mild lesions observed in the low toxicity dosage group marked the safety margin of ANC in SD rats. It was concluded from this study that the toxicity of CaCO3 was dependent on the particulate size (30 ± 5 nm) and concentration and the route of administration used

Preparation and characterization of cockle shell aragonite nanocomposite porous 3D scaffolds for bone repair

The demands for applicable tissue-engineered scaffolds that can be used to repair load-bearing segmental bone defects (SBDs) is vital and in increasing demand. In this study, seven different combinations of 3 dimensional (3D) novel nanocomposite porous structured scaffolds were fabricated to rebuild SBDs using an extraordinary blend of cockle shells (CaCo3) nanoparticles (CCN), gelatin, dextran and dextrin to structure an ideal bone scaffold with adequate degradation rate using the Freeze Drying Method (FDM) and labeled as 5211, 5400, 6211, 6300, 7101, 7200 and 8100. The micron sized cockle shells powder obtained (75 µm) was made into nanoparticles using mechano-chemical, top-down method of nanoparticles synthesis with the presence of the surfactant BS-12 (dodecyl dimethyl bataine). The phase purity and crystallographic structures, the chemical functionality and the thermal characterization of the scaffolds’ powder were recognized using X-Ray Diffractometer (XRD), Fourier transform infrared (FTIR) spectrophotometer and Differential Scanning Calorimetry (DSC) respectively. Characterizations of the scaffolds were assessed by Scanning Electron Microscopy (SEM), Degradation Manner, Water Absorption Test, Swelling Test, Mechanical Test and Porosity Test. Top-down method produced cockle shell nanoparticles having averagely range 37.8±3–55.2±9 nm in size, which were determined using Transmission Electron Microscope (TEM). A mainly aragonite form of calcium carbonate was identified in both XRD and FTIR for all scaffolds, while the melting (Tm) and transition (Tg) temperatures were identified using DSC with the range of Tm 62.4–75.5 °C and of Tg 230.6–232.5 °C. The newly prepared scaffolds were with the following characteristics: (i) good biocompatibility and biodegradability, (ii) appropriate surface chemistry and (iii) highly porous, with interconnected pore network. Engineering analyses showed that scaffold 5211 possessed 3D interconnected homogenous porous structure with a porosity of about 49%, pore sizes ranging from 8.97 to 337 µm, mechanical strength 20.3 MPa, Young's Modulus 271±63 MPa and enzymatic degradation rate 22.7 within 14 days

Synthesis, characterization, and cytocompatibility of potential cockle shell aragonite nanocrystals for osteoporosis therapy and hormonal delivery

Calcium carbonate is a porous inorganic nanomaterial with huge potential in biomedical applications and controlled drug delivery. This study aimed at evaluating the physicochemical properties and in vitro efficacy and safety of cockle shell aragonite calcium carbonate nanocrystals (ANC) as a potential therapeutic and hormonal delivery vehicle for osteoporosis management. Free and human recombinant parathyroid hormone 1-34 (PTH 1-34)-loaded cockle shell aragonite calcium carbonate nanocrystals (PTH-ANC) were synthesized and evaluated using standard procedures. Transmission electron microscopy and field emission scanning electron microscopy results demonstrated highly homogenized spherical-shaped aragonite nanocrystals of 30±5 nm diameter. PTH-ANC had a zeta potential of −27.6 ± 8.9 mV. The encapsulation efficiency of the formulation was found to be directly proportional to the concentrations of the drug fed. The X-ray diffraction patterns revealed strong crystallizations with no positional change of peaks before and after PTH-ANC synthesis. Fourier transform infrared spectroscopy demonstrated no detectable interactions between micron-sized aragonite and surfactant at molecular level. PTH-ANC formulation was stabilized at pH 7.5, enabling sustained slow release of PTH 1-34 for 168 h (1 week). A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cytocompatibility assay in Human Foetal Osteoblast Cell Line hFOB 1.19 showed that ANC can safely support osteoblast proliferation up to 48 h whereas PTH-ANC can safely support the proliferation at 72 h and beyond due to the sustained slow release of PTH 1-34. It was concluded that due to its biogenic nature, ANC is a cytocompatible antiosteoporotic agent. It doubles as a nanocarrier for the enhancement of efficacy and safety of the bone anabolic PTH 1-34. ANC is expected to reduce the cost, dosage, and dose frequency associated with the use of PTH 1-34 management of primary and secondary forms of osteoporosis

In vitro evaluation of a novel pH sensitive drug delivery system based cockle shell-derived aragonite nanoparticles against osteosarcoma

Background: Osteosarcoma (OS) is a highly malignant primary bone cancer. Severe side effects and multidrug resistance are obstacles faced with chemotherapy against OS. With the hope to overcome the obstacles of the conventional chemotherapy, various targeted drug delivery systems using nanotechnology have been explored in the past few decades. Biogenic calcium carbonate (CaCO3) has great potential to be a smart drug delivery system. Results: In this study, cockle shells-derived aragonite nanoparticles (ANPs) were developed and loaded with doxorubicin (DOX). The physicochemical properties of the DOX-loaded ANPs (DOX-ANPs) were characterised by various techniques. The results of drug-loading study demonstrated that DOX was loaded onto ANPs at high loading and encapsulation efficiency (11.09% and 99.58%, respectively). The pH-sensitive release of DOX from DOX-ANPs was successful. At lower pH values (4.8), the release of DOX was much quicker than that at pH 7.4. Additionally, cellular uptake study using fluorescence microscopy showed obviously cellular uptake of DOX-ANPs through endocytosis. Moreover, the flow cytometric analysis revealed DOX-ANPs-induced cell cycle arrest, which was consistent with the mechanism of DOX. DOX-ANPs also showed an efficient cytotoxicity against OS cancer cells, close to the toxicity effect of free DOX at the same concentration. Morphological observations showed microvilli disappearance, chromatin condensation, cell shrinkage, membrane blebbing, and formation of apoptotic bodies, which confirmed both DOX-ANPs- and DOX-induced apoptosis of OS cancer cells in vitro. Conclusion: Our findings indicated that ANPs could act as a pH-sensitive drug delivery against OS

Formulation of a sustained release docetaxel loaded cockle shell-derived calcium carbonate nanoparticles against breast cancer

Purpose: Here, we explored the formulation of a calcium carbonate nanoparticle delivery system aimed at enhancing docetaxel (DTX) release in breast cancer. Methods: The designed nano- anticancer formulation was characterized thorough X-ray diffraction (XRD), Fourier transformed infrared (FTIR), transmission electron microscopy (TEM) and field emission scanning electron microscopy (FESEM) and Brunauer-Emmett-Teller (BET) methods. The nano- anticancer formulation (DTX- CaCO3NP) was evaluated for drug delivery properties thorough in vitro release study in human body simulated solution at pH 7.4 and intracellular lysosomal pH 4.8. Results: Characterization revealed the successful synthesis of DTX- CaCO3NP, which had a sustained release at pH 7.4. TEM showed uniformly distributed pleomorphic shaped pure aragonite particles. The highest entrapment efficiency (96%) and loading content (11.5%) were obtained at docetaxel to nanoparticles ratio of 1:4. The XRD patterns revealed strong crystallizations in all the nanoparticles formulation, while FTIR showed chemical interactions between the drug and nanoparticles with negligible positional shift in the peaks before and after DTX loading. BET analysis showed similar isotherms before and after DTX loading. The designed DTX- CaCO3NP had lower (p 0.05) effects at 48 h and 72 h. However, the DTX- CaCO3NP released less than 80% of bond DTX at 48 and 72 h but showed comparable effects with free DTX. Conclusions: The results showed that the developed DTX- CaCO3NP released DTX slower at pH 7.4 and had comparable cytotoxicity with free DTX at 48 and 72 h in MCF-7 cells

Blood parameter and histopathological changes in osteoporotic rat model treated with cockle shell - derived calcium carbonate nanocrystals

Osteoporosis is a chronic systemic metabolic condition characterized by decreased density of normally mineralized bone due to faulty remodeling process. The success of age long studies on the prevention and treatment of osteoporosis have always been hindered by the ineffectiveness and hazards of the therapeutics and routes of administration used. Nanotechnology is poised to address these issues. This study was aimed at investigating the effectiveness of cockle shell-derived CaCO3 nanocrystals aragonite polymorph as a therapeutic and a hormonal-carrier in the management of primary osteoporosis. Standard techniques were used in the synthesis and evaluations of physicochemical and in vitro/in vivo potentials and safety of ANC and the human recombinant parathyroid hormone (PTH 1-34) - loaded ANC (PTH-ANC) for the management of primary osteoporosis.Transmission Electron Microscopy (TEM) and Field Emission Scanning Electron Microscopy (FESEM) results demonstrated highly homogenized spherical-shaped aragonite nanocrystals of 30±5 nm in diameter. PTH-ANC had a Zeta potential of -27.6 ± 8.9 mV. The encapsulation efficiency of the formulation were found to be directly proportional to the concentrations of the drug fed. The X-ray diffraction (XRD) patterns revealed strong crystallizations with no positional change of peaks before and after PTH-ANC synthesis. Fourier Transform Infrared (FT-IR) spectroscopy demonstrated no detectable interactions between micron aragonite and surfactant at molecular level. PTH-ANC formulation was stabilized at pH 7.5, enabling sustained slow release of PTH 1-34 for 168 hours (one week). A 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) cytocompatibility assay in Human Foetal Osteoblast Cell Line hFOB 1.19 showed that ANC can safely support osteoblast proliferation up to 48 hours while PTH-ANC can safely support the proliferation at 72 hours and beyond due to the sustained slow release of PTH 1-34. A14 day acute toxicity and repeat dose 28 day trial studies of ANC in Sprague Dawley rats recorded no mortality. However, significant haematological anomalies, clinical signs and gross and histopathological lesions were recorded in the acute toxicity groups and the high dose (1g/kg body weight) and medium (0.1 g/kg/kg body weigh) toxicity groups of the repeat dose 28 day trial study. The low dose groups (0.01 g/kg body weight) recorded mild. In vivo efficacy evaluation of antiosteoporotic and drug delivery efficacy of ANC and PTH-ANC demonstrated significant interactions between treatments and regimens. Daily administration of ANC or PTH 1-34, and weekly and fortnightly administrations of PTH-ANC, enabled best gains in bone mass density and strength in ovariectomized and orchidectomized rats. These were consistently demonstrated by results from proliferation and resorption proteomic analyses, bone morphometry and densitometry, flexural 3 point biomechanical bending test, serum calcium and phosphorus analyses, bone ash, calcium and phosphorus analyses and immuhistochemistry. It was concluded that due to its biogenic nature, ANC is a biocompatible antiosteoporotic agent with a cheap method of synthesis. It doubles as a nanocarrier for the enhancement of efficacy and safety of the bone anabolic PTH 1-34. ANC will reduce the cost, dosage and dose frequency associated with the use of PTH 1-34 management of primary form of osteoporosis and enable better compliance to its prescriptions

Morphometric Study on the Developing Kidneys of the Prenatal Dromedary (Camelus dromedarius)

A morphometric study was carried out on the developing kidneys of forty (40) apparently normal fetal dromedary, one-humped camel (Camelus dromedarius) sampled at the Metropolitan abattoir, Maiduguri, Nigeria. The fetuses were obtained from the four prenatal developmental phases of the dromedary: 2 – 4 months, 4 – 7 months, 7 – 10 months and 10 – 13 months. The renal length and diameter showed significant progressive increase in sizes (p<0.005 to 0.001), so were the renal cortical and medullary lengths (p<0.001).  The absolute mean weights of the prenatal kidneys showed significant increase across the growth periods (p<0.001). These progressive increases in absolute mean renal weights were without corresponding increases in the relative mean renal weights known with organ system establishment in mammals. Thus, the dromedary was concluded to have a unique pattern of kidney development