The effect of growth factors on induction of mesenchymal stem cell to the skeletal muscle cells.

OBJECTIVES: The objective of this study was to evaluate the effect of IGF1 and FGF2 growth factors on the differentiation of human adipose tissue­derived mesenchymal stem cells (MSCs) into skeletal muscle cells. METHODS: MSCs were divided into four groups. Group I (control group) was incubated only in myogenic medium, group II was incubated by adding 100 ng/ml FGF2 to the myogenic medium, group III was incubated by adding 20 ng/ml IGF1 to the myogenic medium, group IV was incubated by adding 100 ng/ml FGF2 and 20 ng/ml IGF1 to the myogenic medium. Cells dyed with MyoD1 antibodies were analyzed by flow cytometry so as to determine the myogenic differentiation on day 28. RESULTS: It was confirmed that 11.5 % of the control group, 39.2 % of the FGF2 group, 23.1 % of the IGF1 group, and 39.3 % of the IGF1-FGF2 group showed differentiation. CONCLUSION: Our results show that IGF1 and FGF2 have a positive contribution to myogenic differentiation of MSCs. They contribute to the studies related to muscle diseases and their treatment by the fact that growth factors support the feature of regeneration capacity and differentiation (Tab. 2, Fig. 7, Ref. 23).

The investigation of the antitumoral effect of Cornus mas L in mice with ehrlich solid tumor.

AIM: Cornus mas L is commonly used due to its anti-inflammatory, anti-carcinogenic and anti-oxidant properties. In the study, the effects of C. mas L extract on a solid tumor were examined in the Ehrlich solid tumor model developed in Balb/C type mice. METHODS: Ehrlich acid tumor (EAT) cells (1x106 EAT cell) from the stock animal were injected subcutaneously (s.c.) through the nape of the mice. Treatment groups of solid tumor-induced animals received 100 mg/kg and 200 mg/kg of C. mas L extract intraperitoneally (i.p.) for 14 days. RESULTS: Tumor volumes and animal weights were found to be statistically significant compared to the control group (p < 0.05). AgNOR staining was performed in tumor tissues. Statistically significant differences were observed between the groups in terms of TAA/NA ratio (p < 0.05). Immunohistochemical and biochemical parameters were also evaluated. An estimation of tumor proliferation of the lung, liver, brain, kidney, testis and tumor antioxidant parameters viz. lipid peroxidation, reduced glutathione (GSH), glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) was made. CONCLUSIONS: Our study showed that the anti-tumor effect of C. mas L in assisted tumor development with EAT cells, was mediated by the enhancement of oxidative stress with multiple mechanisms (Tab. 6, Fig. 12, Ref. 38).

The detection of curcumins' antitumoral effects via argyrophilic nucleolar organizing region-associated protein synthesis in mice with ehrlich's ascitic carcinoma.

BACKGROUND: Curcumin is a polyphenol compound that has antioxidant, anticancer, anti-inflammatory, anti-hyperlipidemic and antimicrobial effects. Nucleolar-organizing regions are the sites of the gene on chromosomes. The present study was aimed to show the antitumoral effect of curcumin via AgNOR protein synthesis in Ehrlich's ascitic carcinoma (EAC) bearing mice. METHODS: Twenty three mice with EAC were randomly divided into 3 groups as positive control (n = 7), group 2 (n = 8) and 3 (n = 8) treated intraperitoneally with curcumin (25 mg/kg) and (50 mg/kg), respectively. The animals were sacrificed on Day 16, the solid tumors were removed out. Then, total AgNOR area/nuclear area (TAA/NA) and the mean AgNOR number were estimated for each mice. RESULT: Statistically significant differences were determined among the whole groups for TAA/NA ratio (p = 0.000), conversely mean AgNOR number (p = 0.361). When comparingthe two groups; while no difference was determined between the control and curcumin (25 mg/kg) groups (p = 0.061), the significant differences were detected between the control and curcumin (50 mg/kg) groups (p = 0.000) and between curcumin (25 mg/kg) and curcumin (50 mg/kg) groups (p = 0.000) for TAA/NA ratio. However, there was no significant difference for the mean AgNOR number in double comparison of the groups. CONCLUSIONS: The current study showed that curcumin had a crucial function against cancer development. Also, both AgNOR values might be used as biomarkers for detection of the most reliable therapeutic dose selection of cancer treatment (Tab. 3, Fig. 2, Ref. 27).

Effect of interleukin 12 (IL-12) on embryonic development and yolk sac vascularisation.

BACKGROUND: In the recent days there has been an increase in diseases known as "angiogenic diseases" characterized by pathologic vascularisation. In the rat, the development of embryonic vessel starts to occur at 9.5 days of gestation. In mammals, the vascular system starts developing in a very early embryonic stage. The majority of rat embryo circulation system gets complete approximately at 11 - 12 days. Therefore the in vitro study of 9.5 - 11.5-day old embryo culture could be a suitable model to study the effects of angiogenic and antiangiogenic substances on yolk sac vascularisation. In the present study, the effects of Interleukin-12 (IL-12) on the yolk sac vascularisation are investigated during the in vitro embryo culture, where the latter angiogenic factor was added to serum. METHODS: After 48-hour culture period, effects of different doses of IL-12 (50 ng/ml, 100 ng/ml, and 200 ng/ml) were estimated morphologically. RESULTS: According to morphologic scoring system, the total morphologic score, yolk sac diameter, crown rump length, and somite number were retarded in all experimental groups when compared to control. These developmental retardations were statically significant. There was also a poor development in the yolk sac vascularisation and the heart. CONCLUSION: As a result, the IL-12 could cause developmental retardation of embryos owing to its antiangiogenic effect (Tab. 3, Fig. 2, Ref. 39).

Ropivacaine 0.25% is as effective as bupivacaine 0.25% in providing surgical anaesthesia for lumbar plexus and sciatic nerve block in high-risk patients: preliminary report.

Ropivacaine is potentially less cardiotoxic and neurotoxic than bupivacaine. The aim of this study was to compare the effectiveness of ropivacaine 0.25% and bupivacaine 0.25% for surgical anaesthesia and postoperative analgesia during lumbar plexus and sciatic nerve block in high-risk patients. We performed combined lumbar plexus and sciatic nerve blockade on 62 consecutive ASA III or IV patients undergoing unilateral hip or femur surgery. The first 30 patients received bupivacaine (Group 1) and the remaining 32 patients received ropivacaine (Group 2). Perioperative management was otherwise similar The groups were compared for the time of onset of the block, additional analgesics and sedatives required, time from end of surgery to the first analgesic requirement and the need for rescue analgesia. Ninety percent (29/32) of the patients in the ropivacaine group and 86% (26/30) of the patients in the bupivacaine group reached surgical anaesthesia. The time from the end of the surgery to the first analgesic requirement was similar between the two groups (10.3 +/- 5.2 hours for ropivacaine, 11.2 +/- 4.6 hours for bupivacaine). There was no statistically significant difference between the two groups in any of the measured variables (P > 0.05). The results of this preliminary study suggest that ropivacaine 0.25% is as effective as bupivacaine 0.25% when used for blocking lumbar plexus and sciatic nerve in high-risk patients undergoing hip or femur surgery.

Effects of hysterectomy on ovarian morphology and serum FSH level in rats.

OBJECTIVE: To investigate the effects of hysterectomy with ovarian conservation on ovarian histology and FSH plasma level. METHODS: Fifty female Wistar albino rats (30 for hysterectomy and 20 sham operated for control) were used after two complete estrous cycles. Six months after hysterectomy, the blood samples were collected from both the groups to test FSH plasma level and all animals were sacrificed by decapitation to obtain ovaries for histological examination. RESULTS: Histologic study showed that ovaries from control group exhibited many follicles in various stages of development including primary, secondary and tertiary follicles. There were one or two secondary and tertiary follicles but no primary follicle in each histological section of the 6 months after hysterectomy. The ovaries showed that the cortex of ovary completely covered by corpora lutea, and there were many cystic follicles and atretic follicles with few normal follicles. Theca interna is absent in cysts and most of the mural granulose cell population has been depleted. Serum FSH levels showed significantly increase in hysterectomized group compared control after 6 months operation. CONCLUSIONS: The results of present study support previous studies and suggest that hysterectomy with ovarian conservation in young patients could preserve a woman's normal hormonal milieu. The uterus could have controlling on ovulation and hysterectomy might accelerate to initiate of early menopause.

The effect of vascular endothelial growth factor on in vitro embryonic heart development in rats.

In vitro effects of vascular endothelial growth factor (VEGF) on heart development and total embryonic growth were investigated in 84 rat embryos (obtained from nine pregnant females) at 9.5 days of gestation that were cultured in whole rat serum (WRS), in <30 kDa + >50 kDa serum fractions [retenate (R)], and in R + VEGF. After 24-h culture, the embryos from each group were harvested and divided into two groups. One group was analysed morphologically and biochemically to obtain embryo protein content, the second group was serially sectioned and examined by light microscopy. Morphological score, embryo protein content, somite number and crown-rump length of embryos indicated that embryos cultured in R had significant embryonic retardation, whereas the addition of VEGF to R increased embryonic growth and development. The morphological scores for WRS, R and R + VEGF were 57.7 +/- 0.87, 46.6 +/- 1.90 and 52.1 +/- 0.97, somite numbers were 26.5 +/- 0.47, 20.1 +/- 0.63 and 24.4 +/- 0.46, crown-rump lengths were 3 +/- 0.07, 2.4 +/- 0.06 and 2.7 +/- 0.06 mm, and embryo protein contents were 160.5 +/- 7.41, 98.2 +/- 4.81 and 141.1 +/- 10.96 mug per embryo, respectively. The results of histological examination of heart development were similar. The hearts of embryos grown in R were unseptated and tubular. The atrioventricular endocardial cushions were incompletely developed. The addition of VEGF to R improved heart development. There were no gross morphological differences in the cardiac development between embryos grown in WRS and R + VEGF. In both groups, development of the muscular interventricular septum had begun. Development of the atrioventricular cushions was also similar in both groups and had caused narrowing of the atrioventricular canals, but the atrial septation was not observed.

The growth promoting effects of bFGF, PD-ECGF and VEGF on cultured postimplantation rat embryos deprived of serum fractions.

Serum components in which embryos are cultured in vitro are very important for normal embryonic development. In this study, rat serum was fractionated using Macrosep filters to study the effect of a single growth factor. The fractionated serum, both that containing only material greater than 30 kDa molecular weight (> 30 kDa) and that from which material between 30 kDa and 50 kDa had been removed (< 30 kDa+ > 50 kDa), caused significant embryonic growth retardation. Addition of different concentrations of basic fibroblast growth factor (bFGF, 18 kDa), vascular endothelial growth factor (VEGF, 45 kDa) and platelet-derived endothelial growth factor (PD-ECGF, 45 kDa), to fractionated serum (bFGF to > 30 kDa serum and VEGF or PD-ECGF to < 30 kDa+ > 50 kDa serum) partially restored embryonic growth and development according to a morphological scoring system and protein assay. This restoration was clear by all criteria, as well as in yolk sac vascularisation and heart development. The growth promoting effects of all 3 factors were significant but did not reach the level seen in embryos grown in whole rat serum. The effect of these growth factors was also investigated on anembryonic yolk sac development using a concentration for which maximum whole embryonic growth was seen (128 ng/ml bFGF, 1.6 ng/ml VEGF and 4 ng/ml PD-ECGF), and significant anembryonic yolk sac development was found. These findings suggest that the angiogenic factors may have a growth promoting effect on total embryonic development and vascularisation.

Protection by free oxygen radical scavenging enzymes against salicylate-induced embryonic malformations in vitro.

Salicylates are among the oldest and most widely used drugs and are known to lead to foetal death, growth retardation and congenital abnormalities in experimental animals. In this study, the effects of acetyl salicylic acid (ASA), salicylic acid (SAL) and sodium salicylate (NaSAL) on early organogenesis and the interaction of these molecules with free radicals has been investigated. Postimplantation rat embryos were cultured in vitro from day 9.5 of gestation for 48 hr. ASA, SAL and NaSAL were added to whole rat serum at concentrations between 0.1 and 0.6 mg/ml. Also, the lowest effective concentration of ASA for all parameters (0.3 mg/ml) and the same concentration of NaSAL and SAL was added to the culture media in the presence of superoxide dismutase (SOD) (30 U/ml) or glutathione (0.5 micromol/ml). The growth and development of embryos was compared and each embryo was evaluated for the presence of any malformations. When compared to growth of control embryos, the salicylates decreased all growth and developmental parameters in a concentration-responsive manner. There was also a concentration-related increase in overall dysmorphology, including the incidence of haematoma in the yolk sac and neural system, open neural tube, abnormal tail torsion and the absence of fore limb bud. When SOD was added in the presence of ASA, growth and developmental parameters were improved and there was a significant decrease in the incidence of malformations. Addition of SOD also decreased the incidence of malformations in the presence of SAL, but did not effect the growth and developmental parameters of SAL and NaSAL. There was no significant difference between the embryos grown in the presence of these three molecules on the addition of glutathione. The effects of salicylates might involve free oxygen radicals by the non-enzymatic production of the highly teratogenic metabolites 2,3- and 2,5-dihydroxybenzoic acid. An enhanced production of these metabolites in embryonic tissues may be directly related to the increased risk of congenital malformations.

Teratogenicity of edoferon kappa A, a molecule derived from salicylate, in cultured rat embryos: differences from salicylate and interaction with free oxygen radical scavenging enzymes.

The effect of edoferon kappa A (E-KA), a non-specific immunomodulatory and anti-neoplastic chemical substance derived from the methyl form of salicylate (acetyl salicylic acid; ASA), on mammalian embryos was studied and compared to the effects of ASA. Rat embryos were cultured in vitro from 9.5 days of gestation for 48 h. E-KA (0.1-12.8 mg/ml) and ASA (0.1-0.6 mg/ml) were added to the whole rat serum. To investigate the interaction of these molecules with antioxidant agents, the lowest effective concentrations of E-KA (0.6 mg/ml) and ASA (0.3 mg/ml) for all parameters were added to the culture media in the presence of superoxide dismutase (SOD) (30 U/ml) or glutathione (0.5 mumol/ml). The growth and development of embryos was compared and each embryo was evaluated for the presence of any malformations. E-KA and ASA decreased growth and development in a concentration-responsive manner. There was also a concentration-related increase in overall dysmorphology (haematoma in the yolk sac and neural system, open neural tube, abnormal tail torsion and the absence of fore limb bud). There were no statistically significant differences between the control and embryos grown in the presence of 0.1-0.4 mg/ml E-KA, although the effects of ASA started at a concentration of 0.2 mg/ml. Embryos showed significant growth retardation in all scoring criteria and severe malformations when 0.5-3.2 mg/ml E-KA and 0.3-0.6 mg/ml ASA were added. When SOD was added, there was a significant decrease in the incidence of malformations and growth and developmental parameters were increased but this decrease never reached the control level. We concluded that E-KA has direct toxic effects on the developing embryo but at much higher concentrations than ASA, and the teratogenic effects of these molecules might be related to free oxygen radicals.