ABSTRACT
Delayed union and nonunion fractures are clinical challenges for orthopedic surgeons. The development of fracture complications, such as delayed union and nonunion fractures, is still difficult to predict. Various methods are being investigated to improve fracture healing and prevent complications in patients. There are various methods to promote fracture healing, broadly divided into biological, chemical, and physical methods. One of the most widely used physical methods to promote fracture healing is the pulsed electromagnetic field (PEMF). This study aimed to evaluate the healing process of delayed union fracture after being stimulated by PEMF. Twenty-four rats were randomly divided into two groups: the control group (n = 12) and the PEMF group (n = 12). Delayed union fracture was performed on the left femur of all rats. Subsequently, the PEMF group was given PEMF stimulus with a magnetic field intensity of 1.6 mT and a frequency of 50 Hz for 4 hours/day and 7 days/week. The fracture healing process was evaluated on days 5, 10, 18, and 28 based on the bone callus histology using safranin O fast green (SOFG) staining. The results of the histological analysis showed that bone cartilage was higher in the PEMF group than in the control group throughout the observation period. In addition, the PEMF group had less fibrous tissue at the beginning of the healing. This finding indicates PEMF stimulation has an effect on inducing osteogenesis on fracture healing and reducing the risk of delayed union.
ABSTRACT
BACKGROUND: Recently, Phosphatidylcholine (PC) has been used as an off-label treatment for lipolysis injection, which is associated with inflammatory reaction due to sodium deoxycholate, an emulsifier, so that inflammation as side effect occurs in those patients. Liposome formulation from soybean lipid was thought to be a better and safer alternative. This study aimed to analyze the mechanism of Liposomal Soybean Phosphatidylcholine (LSPC) extract from Indonesian soybeans (containing 26% PC) to induce Adipose-derived Stem Cells (ASCs) death in vitro. METHODS: Liposomes were prepared using thin film hydration method followed by a stepwise extrusion process to produce a small amount of 41.0-71.3 nm. Liposomal soybean phosphatidylcholine extract (LSPCE), liposomal purified PC (LPCC), and solution of PC+SD were used for comparison. Annexin V fluorescein Isothiocyanate/Propidium Iodide (FITC/PI) double staining by flow cytometry and also measurement of caspase-3 activity using ELISA were used to quantify the rate of apoptosis. ASCs viability was measured using MTT assay after induction with liposomes. Morphological changes were shown using a phase-contrast, inverted microscope and Transmission-Electron Microscope (TEM). RESULTS: The flow cytometry results showed that cells treated with both LSPCE and LPCC showed increase in early apoptosis beginning at 6 hr after incubation, which was confirmed by caspase 3 measurement. MTT assay showed that both LSPCE and LPCC could decrease viability of cells. Cells treated with LSPCE and LPCC showed some rounded cells, which was an early sign of cell death. Cells treated with SD showed extensive membrane damage with necrosis features using TEM. CONCLUSION: The results above demonstrated that LSPCE induced apoptosis of ASCs.
ABSTRACT
BACKGROUND: Epididymal sperm maturation occurs via interactions between sperm and proteins secreted by the epididymal epithelium. Although this is an important process, the genes that encode the involved proteins remain largely uncharacterized. Previous studies have demonstrated that the genes involved in sperm maturation are regulated by androgen. Spag11a is an epididymal gene that is influenced by androgen. However, little is known about the putative role of this gene in the sperm maturation process. The objective of this study was to characterize Spag11a in the mouse epididymis. METHODS: In silico analyses were performed to predict signal peptides and functional domains. Spag11a expression was measured by quantitative real-time RT-PCR. Western blots and immunocytochemistry were performed to determine protein expression. RESULTS: SPAG11A is a member of the beta defensin protein family and constitutes a secretory protein. Spag11a was expressed exclusively in the epididymis. Moreover, it exhibited region-specific expression in the caput, which is typical for genes that are involved in creating a suitable microenvironment for sperm maturation. Mouse Spag11a was regulated by androgen. A significant decrease of Spag11a expression was observed at third day following a gonadectomy (P < 0.001). Interestingly, testosterone replacement therapy was able to maintain the expression almost at the normal level, indicating a dependency on androgen. Besides androgen, testicular factors influenced Spag11a expression in a different way. This was revealed by efferent duct ligation in which Spag11a was transiently up-regulated at the third day following the ligation before returning to the normal level at day 5. Spag11a regional expression was also observed at protein level detected by western immunoblotting which revealed a clear band in the caput but not in other regions. The prediction that SPAG11A is a secretory protein was confirmed by immunocytochemical analyses indicating cell-specific expression mainly in the caput principal cells and detection of the protein in epididymal luminal fluid and spermatozoa. CONCLUSIONS: Based on the characteristics of Spag11a, it is likely that this gene has a specific role in epididymal sperm maturation. Further studies using functional assays are necessary to confirm this finding.
