Evaluation of the restorative effect of ozone and chitosan-hyaluronic acid with and without mesenchymal stem cells on wound healing in rats.

This study evaluated the effect of ozone, chitosan-hyaluronic (Cs-HA) acid and mesenchymal stem cells (MSCs) on wound healing in rats. A total of 64 rats were randomly divided into four groups: control, ozone, Cs-HA + ozone and Cs-HA + ozone + MSCs. A 5 mm full-thickness wound was created on the back of each rat. The wound area was measured macroscopically on days 3, 5, 9 and 14. Tissue sections were prepared for histopathological evaluation of inflammation, collagen arrangement, neovascularization and epithelial tissue rearrangement. Macroscopic assessment showed differences in wound area on days 5, 9 and 14. Histopathological examination showed that the Cs-HA + ozone + MSCs and Cs-HA + ozone groups had significantly higher vascularization on day 3 compared to the ozone-treated and control groups. All treatment groups had significantly better collagen arrangement than the control group. On day 5, no significant difference was observed between different groups. On day 9, the inflammation level in the Cs-HA + ozone + MSCs group was significantly lower than in the other groups. All treatment groups had significantly better vascularization compared to the control group. On day 14, the rate of inflammation was significantly lower in the treatment groups than in the control group. Significantly higher collagen arrangement levels were observed in the Cs-HA + ozone and Cs-HA + ozone + MSCs groups compared to the control and ozone groups. All treatment groups had significantly better epithelial tissue rearrangement than the control group. Overall, the results of this study indicated that treatment with ozone, Cs-HA acid, Cs-HA and MSCs accelerated wound healing in rats. The effect of using Cs-HA acid with mesenchymal cells was better than the other types of treatment. Larger clinical trials are needed to assess these factors for improving chronic wound treatment.

The healing effect of a mixture of Arnebia euchroma and animal fat on burn wounds in rats in comparison with sulfadiazine.

OBJECTIVE: Thermal burn is a serious cause of morbidity and mortality that affects millions of people worldwide. The aim of this experimental study was to investigate the efficacy of Arnebia euchroma (AE) to treat burn wounds in a rat model. METHOD: A total of 80 male rats (200-250g) were shaved over the back of the neck (2×3cm2) and a second-degree burn wound was induced at this site under general anaesthesia. The rats were then randomly assigned to one of four groups (each n=20) and the burns were treated daily for 14 days as follows: (1) dressed with animal fat; (2) dressed with sulfadiazine; (3) dressed with a mixture of AE and animal fat; (4) no treatment (control). Five rats from each group were sacrificed on days 3, 5, 9 and 14 post-burn and the wounds were evaluated histologically and immunohistochemically for the expression of interleukin (IL)-1 and IL-6. RESULTS: There was a significant increase at day 3 and decrease on day 5 samples for the expression of IL-1 in the AE plus fat group and IL-6 in the AE plus fat and sulfadiazine groups, compared to the control and fat treatment groups, respectively. Both AE plus fat and sulfadiazine treatments reduced inflammation and granulation tissue formation by day 5 post-burn, while re-epithelialisation commenced by day 9 post-burn. In addition, burns treated with AE plus fat exhibited keratinised epidermis, associated with regular collagen fibres, compared to moderately dense collagen fibres without vascularisation in the sulfadiazine group. CONCLUSION: These findings suggested that AE plus fat was superior to sulfadiazine in enhancing burn wound healing in rats.

Combined treatment with Alhagi maurorum and docetaxel inhibits breast cancer progression via targeting HIF-1α/VEGF mediated tumor angiogenesis in vivo.

Breast cancer is a challenging disease and leading cause of cancer death in women. There is no effective agent for metastatic breast cancer after surgery and chemotherapy. Alhagi maurorum (A.m) has been reported to exhibit an anticancer effect on various types of cancer cells in vitro. This study aimed to examine the suppressive effect of A.m alone and combined with docetaxel (DTX) on the breast cancer growth in mice models and the possible underlying mechanisms. In the present study, the mice were inoculated subcutaneously with the injections of 4T1 cells. Then, A.m, DTX, and their combination were administered intraperitoneally. The expressions of ß-catenin (ß-cat), FZD7, MMP2, HIF1-α, and VEGF A (vascular endothelial growth factor A) were investigated using RT-PCR method. Also, plasma alkaline phosphatase (ALP), alanine aminotransferase (GPT or ALT), aspartate transaminase (GOT or AST), serum creatinine, and urea were examined, and histological analyses of the tissues were conducted. The results demonstrated that A.m (500 mg/kg) combined with DTX significantly decreased the expression of ß-cat, MMP2, and FZD7 as compared with the negative control group and monotherapies. Also, the mRNA levels of HIF1-α and VEGF A were suppressed significantly by DTX + A.m (500 mg/kg). Tumor weights and sizes were significantly lower and tumor inhibition rate was significantly higher in the DTX + A.m group. The A.m 500 mg/kg + DTX also suppressed the serum GPT level in tumor-bearing mice and decreased the serum urea level. Taken together, our findings suggest that DTX combined with A.m at an optimal dose of 500 mg/kg as the optimal dose can inhibit ß-cat, FZD7, MMP2, and breast cancer growth via interrupting HIF-1α/VEGF signaling and might be used as a promising antiangiogenic agent for breast cancer treatment.

Effect of pancreatin on acute pancreatitis resulting from L-arginine administration in mice, a morpho-histopathological and biochemical study

Abstract Acute pancreatitis (AP) is a life-unpleasant situation with contradictory and inadequate treatments. In this regard, the present study evaluated the effect of the possible pretreatment of lipase-pancreatin on L-arginine-induced AP. Forty adult mice were selected and divided into five groups: I) control group, II and III) AP groups (i.p.) receiving L-arginine of 2×300 and 2×400 mg/100 g body weight (b.w.), IV) AP (2×300 L-arginine) group + pancreatin (mice were i.p. injected by 350 U-lipase), and V) AP (2×400 L-arginine) group + pancreatin (mice were i.p. injected by 350 U-lipase). All AP groups displayed a significant increase in serum levels of ALT, AST, TBARS, and TNF-alpha compared to the control group. Moreover, pancreatic tissue edema, inflammation, and vacuolization of acinar cells were significantly higher in the untreated L-arginine group compared to the control and pancreatin groups. Conversely, the diameter of pancreatic islets significantly declined after induction of pancreatitis compared with control and pancreatin groups. Pancreatin treatment can be used in pancreatic dysfunction, however, this medicine showed no protective effect against L-arginine-induced AP in the mouse model.

The effect of iron sulfate nanoparticles and their fortified bread on Wistar rats and human cell lines.

BACKGROUND: Ferrous sulfate nanoparticles (FSNPs) were synthesized and characterized to mitigate the undesirable effects of ferrous sulfate bulk particles (FSBPs) as a supplement or fortificant in health/food industries. METHODS: The toxicity of FSNPs and FSBPs was evaluated against AGS, PLC/PRF/5, and HGF1-PI 1 cell lines. Then, Wistar rats were fed three levels of FSNPs and FSBPs fortified-bread. Growth performance, hematological parameters, and histopathological changes in treated rats were assessed after 21 days. RESULTS: High concentrations of FSNPs (3.125 and 6.25 mM) increased the necrosis of AGS cells. A low level of FSNPs (1.57 mM) did not affect the viability of cells after 72 h. Fibroblasts did not show apoptosis and necrosis after exposing 1.57 mM of FSNPs. In rats, 9 mg elemental iron of FSNPs/day enhanced hemoglobin, PCV, and ferritin values and increased the body weight gain (p < 0.05). FSNPs fortified-bread induced no clinical symptom or histopathological lesion in rats. CONCLUSION: FSNPs affect cells in a dose-dependent manner. The results indicate that FSNPs at the low level do not have adverse effects on normal fibroblasts and rats. Significant weight gain in rats having a low level of FSNPs compared to the FSBPs indicates the negligible toxicity of FSNPs at low concentrations.

Comparison Capacity of Collagen Hydrogel and Collagen/Strontium Bioglass Nanocomposite Scaffolds With and Without mesenchymal Stem Cells in Regeneration of Critical Sized Bone Defect in a Rabbit Animal Model.

Bone self-healing is limited and requires additional or external intervention to promote and accelerate bone regeneration. Therefore, the aim of this study was to investigate the potential capacity of hydrogel collagen (Co) nanocomposite alone, and in combination with 2% strontium (Co/BGSr2%) in presence of mesenchymal stem cells (MSCs) in full-thickness bone defect regeneration in the rabbit animal model. A total of 72 New Zealand white rabbits were randomly divided in 6 groups of 12 rabbits with full-thickness bone defect. In five groups, the bone defect was treated with MSC, Co, Co/BGSr2%, Co + MSCs, and Co/BGSr2% + MSCs. No treatment was done in the control group. The treatments were assessed radiographically, histopathologically, and immunohistochemically on days 14, 28, 42, and 56 post-treatment. The highest radiographical and histological scores were belonged to the Co/BGSr2% + MSC followed by Co + MSCs, Co/BGSr2%, Co, MSC, and the control groups. The highest and lowest mean expression level of osteocalcin was detected in the Co/BGSr2% + MSC and control groups by 28th dayof post-implantation, respectively. In contrast, the highest and lowest mean expression level of osteocalcin on day 56 post-implantation was belonged to the control and Co/BGSr2% + MSC, respectively. The Co/BGSr2% nanocomposite scaffold seeded with MSC can accelerate bone regeneration resulted from osteoblastic production of osteocalcin protein. Therefore, collagen hydrogel combined with 2% strontium in nanocomposite form is a suitable candidate scaffold for bone tissue engineering.

Comparison capacity of collagen hydrogel, mix-powder and in situ hydroxyapatite/collagen hydrogelscaffolds with and without mesenchymal stem cells and platelet-rich plasma in regeneration of critical sized bone defect in a rabbit animal model.

The aim of this study was to investigate the effect of developed collagen (Co) hydrogel (CH), powder-mixed hydroxyapatite/collagen (HA/Co) hydrogel and in situ synthesized HA/Co (In/HA/Co) hydrogel with or without mesenchymal stem cell (MSC) and platelet-rich plasma (PRP) on the regeneration of full-thickness critical size bone defect in the rabbit animal model. In the first step of this study, the scaffolds were synthesized and characterized using FTIR spectroscopy, X-ray diffraction, and scanning electron microcopy. In the second step or animal study, the radial bone defects were filled with the synthesized scaffolds with and without MSC and PRP. One hundred sixty one year-old New Zealand white male rabbits were randomly divided in 16 groups of 10 rabbits including control with bone defect without treatment, In/HA/Co, HA/Co, CH, PRP, MSC, CH + PRP, HA/Co, In/HA/Co + PRP, HA/Co + PRP, CH + MSC, In/HA/Co + MSC, HA/Co + MSC, CH + PRP + MSC, In/HA/Co + PRP + MSC, and HA/Co + PRP + MSC. The created defects were filled using the constructed scaffolds alone or seeded with MSCs, with and without PRP injection. The treatments were assessed using histopathological, immunohistochemical and rediographical analysis on days 14, 28, 42, 56 post-treatment. The plate-like HA particles were distributed homogeneously in the in situ HA/Co scaffold compared to the HA/Co scaffold and had a similar structure to bone with carbonated plate-like HA particles and nanofibrilated Co matrix. In situ HA/Co nanocomposite seeded with MSC and enriched by PRP can accelerate bone regeneration resulted from osteoblastic production of osteocalcin protein. Therefore, in situ HA/Co hydrogel seeded with MSC and PRP can be a new approach for bone tissue engineering.

Effect of White Tea (Camellia sinensis) Extract on Skin Wound Healing Process in Rats.

BACKGROUND: White tea (Camellia sinensis) has anti-inflammatory and antioxidant properties and a protective effect against wrinkles, sunburn and UV damages on the skin. Thus, we aimed to evaluate the effect of white tea extract on the healing process of skin wounds in rats. METHODS: This study was done in the Research Center of Shahrekord University of Medical Sciences, Shahrekord, Iran in 2019. Excisional skin wounds were created on five groups of healthy male Wistar rats (200-250 g, n=21) including control group, Eucerin-treated group, white tea 5% ointment (Eucerin) treated group, gel-treated group, white tea 5% gel treated group. Treatment was begun on day 1 and repeated every day at the same time until day 15. Pathologic samples were taken on days 4, 7 and 15 for histopathological examinations. Kruskal-Wallis test was used to analyze data by SPSS. Statistical significance was defined as P<0.05. RESULTS: Wound closure rate of control group was more than other groups on day 4 (P<0.05). On day 7, reepithelisation and granulation tissue of control group were more than white tea 5% ointment-treated and its inflammation was less than others (P<0.05). Neo-vascularization of white tea 5% ointment-treated group was more than control group on days 4 and 15 (P<0.05). On day 4, intact mast cells of control group were more than white tea treated groups (P<0.05). Degranulated mast cells of white tea 5% gel treated group was significantly (P<0.05) more than control group on days 4 and 15. CONCLUSION: Five percent white tea extract could not help the skin wound healing process.

Polyethersulfone/MWCNT nanocomposite scaffold for endometrial cell culture: preparation, characterization, andin vitroinvestigation.

Endometrial cell culture is a method for investigating physiological or pathological conditions or simulatingin vivoconditions for embryo culture. The natural function of the endometrium depends on a polarized epithelium and 3D stromal compartments. The polymer-based scaffolds of simple polyethersulfone (PES), laser scratched PES (PES-LS), and multiwall carbon nanotubes (MWCNT) composited PES (PES-MWCNT) were prepared and used for bovine endometrial cells (bECs) culture. For better investigation of the relationship between physical structure and cell growth behavior, the surface morphologies of the scaffolds were evaluated by atomic force microscopy (AFM) and field emission scanning electron microscopy (FESEM) techniques. Three synthesized membranes (PES, PES-LS, and PES-MWCNT) were evaluated for the cell morphology, viability and, doubling time. Results showed acceptable physical and chemical fabrication of the polymers with no significant differences in the proportions of live cells to primary cultured cells, dead to live cells, and the cell doubling time among groups during the experiment (P > 0.05). Total cell count (live and dead cells) was significantly different on Day 2 among types of polymers. The results showed the comparable potential of the PES-MWCNT membrane for the bECs culture.

Taenia ovis in Small Ruminants in Iran: Prevalence, Pathology, and Economic Loss.

Taenia ovis larvae can result in economic losses in small ruminants due to condemnation of infected tissues or whole carcasses. From 2017 to 2018, the T. ovis prevalence in 16,180 sheep and 7560 goats at the Najafabad slaughterhouse in Isfahan was determined. More sheep (477; 2.9%) than goats (90; 1.2%) were found to be infected, and the prevalence was higher in animals <1 y (p < 0.0001), and higher in spring in sheep (8.2%) and goats (2.2%). Female sheep were more frequently infected than males (p < 0.0001); this did not hold true for goats. Of the tissues examined, T. ovis was found more often in the heart muscle of sheep compared with other tissues; however, infections in the heart muscle, masseter muscle, diaphragm, and triceps were similar in goats. Granulomas and caseous necrosis in the heart muscles were associated with the accumulation of mononuclear inflammatory cells and the formation of fibrous tissue around the parasite. Based solely on infected tissues found in this study, the economic loss caused by the presence of T. ovis larvae was estimated to be 4167 United States dollars (USD). Control methods, such as proper disposal of infected tissues and anthelmintic treatment of infected dogs, are necessary to decrease infection and prevent economic loss in small ruminants.

17ß-estradiol improves the efficacy of exploited autologous bone marrow-derived mesenchymal stem cells in non-union radial defect healing: A rabbit model.

Exploiting mesenchymal stem cells (MSCs) appears to be an appealing alternative to the traditional clinical approach in the treatment of non-union bone defects. It has been shown that 17ß-estradiol improves the osteogenesis and proliferation potential of the MSCs via estrogen receptors. We investigated the effect of 17ß-estradiol on exploiting autologous BMSCs (bone marrow-derived MSCs) for the purpose of healing of radial non-union segmental defect in rabbit. Twenty rabbits were divided into 4 experimental groups: 1. Control group; 2. MSC treatment group; 3. 17ß-estradiol (E2) treatment group; and 4. E2+MSC treatment group. Isolated BMSCs were seeded in a critical-sized defect on radial mid-diaphysis that was filled with autologous fibrin clot differently in 4 groups: 1. intact fibrin clot (control); 2. Fibrin clot containing MSCs; 3. Estradiol; and 4. E2 and MSCs. Defect healing was assessed by radiological (week 0, 2, 4, 6, 8 and 10) and histopathological evaluation (week 10). Radiological evaluation data demonstrated that quantities for the E2+MSC group were significantly the greatest in comparison with the other groups at week 4 to 10 inclusive. Moreover, Histopathological evaluation indicated that the E2+MSC group had the highest score which was significantly greater than the E2 group and the control group (P<0.05). In-vivo application of in situ 17ß-estradiol provides the seeded BMSCs with improved osteogenic capacity in tandem with an accelerated rate of bone healing. This obviously more qualified approach that yields in a shorter time appears to be promising for the future cell-based clinical treatments of the non-union bone fractures. Exploiting mesenchymal stem cells (MSCs) appears to be an appealing alternative to the traditional clinical approach in the treatment of non-union bone defects. It has been shown that 17ß-estradiol improves the osteogenesis and proliferation potential of the MSCs via estrogen receptors. We investigated the effect of 17ß-estradiol on exploiting autologous BMSCs (bone marrow-derived MSCs) for the purpose of healing of radial non-union segmental defect in rabbit. Twenty rabbits were divided into 4 experimental groups: 1. Control group; 2. MSC treatment group; 3. 17ß-estradiol (E2) treatment group; and 4. E2+MSC treatment group. Isolated BMSCs were seeded in a critical-sized defect on the radial mid-diaphysis that was filled with autologous fibrin clot differently in 4 groups: 1. intact fibrin clot (control); 2. Fibrin clot containing MSCs; 3. Estradiol; and 4. E2 and MSCs. Defect healing was assessed by radiological (week 0, 2, 4, 6, 8 and 10) and histopathological evaluation (week 10). Radiological evaluation data demonstrated that quantities for the E2+MSC group were significantly the greatest in comparison with the other groups at week 4 to 10 inclusive. Moreover, Histopathological evaluation indicated that the E2+MSC group had the highest score which was significantly greater than the E2 group and the control group (P<0.05). In-vivo application of in situ 17ß-estradiol provides the seeded BMSCs with improved osteogenic capacity in tandem with an accelerated rate of bone healing. This obviously more efficient approach that yields in a shorter time appears to be promising for future cell-based clinical treatments of the non-union bone fractures.

Spinal fusion with demineralized calf fetal growth plate as novel biomaterial in rat model: a preliminary study.

BACKGROUND: Spinal fusions are being performed for various pathologies of the spine such as degenerative diseases, deformities, tumors and fractures. Recently, other bone substitutes such as demineralized bone matrix (DBM) have been developed for spinal fusion. Therefore, this study was conducted to evaluate the intertransverse posterolateral fusion with the Bovine fetal growth plate (DCFGP) and compare it with commercial DBM in rat model. METHODS: A total of 16 mature male rats (aged 4 months and weighing 200-300 g) were randomly divided in two groups. After a skin incision on posterolateral site, two separate fascial incisions were made 3 mm from the midline. A muscle-splitting approach was used to expose the transverse processes of L4 and L5. Group I (n = 8) underwent with implanted Bovine fetal growth plate among decorticated transverse processes. In group II (n = 8) commercial DBM was placed in the same manner. Fusion was evaluated by manual palpation, radiographical, gross and histopathological analysis. RESULTS: The manual palpation, radiological, gross and histopathological findings indicate high potential of the DCFGP in spinal fusion. At the 42nd postoperative day, new bone formation as evidenced by a bridge between L4 and L5 was visualized in all rats implanted with DCFGP and commercial DBM. The newly formed bone tissue was observed in all implanted areas on the 42nd day after operation in the two groups. CONCLUSIONS: The spinal fusion of the animals of both groups demonstrated more advanced osteogenic potential and resulted in proper fusion of the transverse process of lumbar vertebra.

Effects of adipose tissue stem cell concurrent with greater omentum on experimental long-bone healing in dog.

Repair of large bone defects resulting from trauma, tumors, and osteitis is a current challenge to surgeons. Adipose-derived adult stem cells (ASCs) are multipotent cells that are able to differentiate into osteoblasts in the presence of certain factors. In this study, the role of greater omentum as a scaffold incorporation of ASCs was evaluated in long-bone defect healing in dog model. Sixteen 3-4-year-old, male adult mongrel dogs, weighing 25.2 ± 3.5 kg, were used in this study. In the control group (n = 4), the defect was left empty. In the omental group (n = 4), the defect was filled with harvested omentum. In the omental-ASCs group (n = 4), the defect was filled with omentum and 1 mL of ASCs was injected into the grafted omentum. In the omental-culture medium group (n = 4), 1 mL of culture medium was injected into the grafted omentum. Finally, the injured radial bones were fixed with plate and screw. Radiographs of each forelimb was taken postoperatively on the first day and at the second, fourth, sixth, and eighth weeks postinjury to evaluate bone formation, union, and remodeling of the defect. The operated radii were removed on the 56th postoperative day and were histopathologically evaluated. In this study, both omental-culture medium and omental-ASCs groups demonstrated superior osteogenic potential in healing the radial bone defect. Compared to those of the omental and control groups, more advanced bone healing criteria were present in the omental-culture medium and omental-ASCs groups at radiological and histopathological levels at 8 weeks postsurgery.

Effects of captopril on the cysteamine-induced duodenal ulcer in the rat.

Oxidative stress is important factor underlying in a variety of diseases. Antioxidative enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) are part of the physiological defenses against oxidative stress. Malondialdehyde (MDA) is a lipid peroxidation biomarker and its elevated level in various diseases is related to free radical damage. Cysteamine is a cytotoxic agent, acting through generation of reactive oxygen species (ROS) and may decrease defense activity of antioxidative enzymes against ROS and induce duodenal ulcer. Captopril, acts as free radical scavengers and protect against injuries from oxidative damage to tissues.The aim of this study was the evaluation of the effect of captopril against cysteamine-induced duodenal ulcer by determining duodenal damage, duodenal tissue SOD and GSH-PX activities and plasma MAD level. This study was performed on 3 groups of 7 rats each: saline, cysteamine and cysteamine plus captopril treated groups. The effect of captopril against cysteamine-induced duodenal ulcer is determined by evaluating the duodenal damage, duodenal tissue SOD and GSH-PX activities and plasma MDA level. All animals were euthanized 24h after the last treatment and 2 ml blood and duodena samples were collected for calculation of ulcer index, histopathological assessment and measurement of tissue SOD, GSH-PX activities and plasma MDA level. Cysteamine produced severe duodenal damage, decreased the activity of duodenal tissue SOD and GSH-PX and increased the plasma MDA level compared with saline pretreated rats. Pretreatment with captopril decreased the cysteamine-induced duodenal damage and plasma level of MDA and increased the activities of SOD and GSH-PX in duodenal tissue compared with cysteamine pretreated animal. Our results suggest that captopril protects against cysteamine-induced duodenal ulcer and inhibits the decrease in SOD and GSH-PX activities and lipid peroxidation by increasing antioxidant defenses.