Effects of potassium apigenin and verbena extract on the wound healing process of SKH-1 mouse skin.

Tissue repair is a complex process, which may be favoured or inhibited by different factors. Potassium apigenin (AP) and other flavonoids present in verbena extract (PLX(®) ) possess powerful anti-inflammatory properties. The aim of this study was to evaluate the effects of topical treatment with AP and PLX gels on wounds inflicted on SKH-1/CRL mice. Forty-eight SKH-1 mice were used (4 groups with 12 animals each), which were subjected to wound excision with a round scalpel, 4 mm in diameter, on the dorsal skin. The animals were divided into four groups: Group I received topical applications of apigenin gel; Group II received PLX gel; Group III received vehicle gel; Group IV acted as control. Wound contraction, reepithelialisation, inflammation and neovascularisation (by means of immunohistochemical staining with anti-laminin) were recorded at study periods established at 2, 7 and 14 days. Reepithelialisation was faster in Groups I and II at 7 days (56·25% grade 3 and 43·75% grade 4) compared with the other groups. The degree of inflammation showed improvement with a tendency towards statistical significance in Groups I and II at 2 and 7 days. Anti-laminin staining was more intense in the group treated with PLX at the 2- and 7-day periods. Topical treatment with PLX gel improved the degree of reepithelialisation and inflammation, and favoured neo-vascularisation of the wounds at 2 and 7 days following surgery.

Bone response to hydroxyapatites with open porosity of animal origin (porcine [OsteoBiol mp3] and bovine [Endobon]): a radiological and histomorphometric study.

PURPOSE: To carry out a radiological and histomorphometric evaluation of bone response to two xenografts of animal origin, one porcine, and the other bovine, inserted in rabbits' tibiae. MATERIAL AND METHODS: Twenty New Zealand rabbits weighing 3900-4500 g were used. Twenty bovine bone grafts (Endobon) in granulated form of 500-1000 µm granulometry were inserted in the proximal metaphyseal area of the animals' right tibia, and 20 porcine bone grafts (OsteoBiol mp3) in granulated form of 600-1000 µm granulometry were inserted in the proximal metaphyseal area of the animals' left tibia. Following graft insertion, the animals were sacrificed in four groups of five, after 1, 2, 3 and 4 months, respectively. Anteroposterior and lateral radiographs were taken. Samples were processed for observation under light microscopy. Histomorphometric measurements were presented as mean values ± standard deviations. RESULTS: At 4 months after treatment, the bone defects displayed radiological images that showed complete repair of osseous defects. Histomorphometric evaluation showed that for the porcine xenograft, the study averages for newly formed bone represented 22.8 ± 1.8%, for residual graft material 23.6 ± 3% and for connective tissue 53.5 ± 2.5%, while for the bovine xenograft newly formed bone represented 23.1 ± 1.8%, residual graft material 39.4 ± 3% and non-mineralized connective tissue 37.5 ± 2.5%. CONCLUSIONS: The biomaterials assessed in the study were shown to be biocompatible and osteoconductive. Collagenized porcine xenografts proved more resorbable than bovine xenografts. Both can be used as possible bone substitutes without interfering with normal reparative bone processes.

Perioperative antibiotic regimen in rats treated with pamidronate plus dexamethasone and subjected to dental extraction: a study of the changes in the jaws.

PURPOSE: Osteonecrosis of the jaws is a well-known condition associated with long-term bisphosphonate use. This study analyzed the prophylactic effect of antibiotic treatment in Sprague-Dawley rats treated with pamidronate plus dexamethasone and subjected to oral surgery in the form of dental extraction. MATERIALS AND METHODS: One hundred twenty animals were included in a randomized prospective study. Animals in group I (n = 60) were treated with dexamethasone 1 mg/kg and pamidronate 3 mg/kg 3 times per week for 7, 14, and 21 days. All were subjected to right mandibular or maxillary molar extraction 8, 15, and 22 days after the start of dosing. Animals in group II (n = 60) received the same treatment except for the addition of penicillin 0.1 mL/kg per day for 3 days before and 4 days after extraction. Rats in the 2 groups were sacrificed 14 and 28 days after extraction. A clinical and histologic evaluation was performed. RESULTS: In group I, osteonecrosis was documented in 18 cases (34.6%; affecting the upper and lower jaws in 10 and 8 cases, respectively). In group II, osteonecrosis was documented in 5 cases (9.61%; affecting the upper and lower jaws in 3 and 2 cases, respectively). The difference between the 2 groups was statistically significant (P = .002). CONCLUSIONS: The adoption of preventive measures (antibiotic prophylaxis) in invasive dental procedures results in a significant decrease in osteonecrosis of the jaws associated with bisphosphonate use.

An experimental study of bisphosphonate-induced jaws osteonecrosis in Sprague-Dawley rats.

OBJECTIVE: Patients undergoing treatment with bisphosphonates may develop jaw lesions consisting mainly of bone necrosis. The present study describes a model of maxillary osteonecrosis in Sprague-Dawley rats, applying bisphosphonates and examines the changes occurring after tooth extraction. MATERIAL AND METHODS: A total 200 animals were included in a randomized prospective study involving the following groups: group I (control, 20 rats without drug treatment), group II (60 animals administered dexamethasone 1 mg/kg/day for 7, 14 and 21 days, in subgroups of 20 animals each), group III (60 animals administered pamidronate daily at a dose of 3 mg/kg) and group IV (60 animals administered pamidronate and dexamethasone). In all groups, molar extraction was carried out on the right upper maxillary or mandibular side 8, 15 and 22 days after the start of dosing. The rats were killed 14 and 28 days after extraction in all groups. RESULTS: A total of 18 cases of osteonecrosis were recorded in the group administered pamidronate and dexamethasone. Osteonecrosis affected the upper maxilla in 10 cases and the mandible in eight cases, and was circumscribed to the extraction zone in all cases. Osteonecrosis was not seen in any of the other groups. CONCLUSIONS: The administration of pamidronate and dexamethasone in rats subjected to molar extraction increases the risk of osteonecrosis.

Effects of plasma rich in growth factors on wound healing of the tongue. Experimental study on rabbits.

OBJECTIVES: To apply autologous Plasma Rich in Growth Factors (PRGF) in wounds provoked in the tongue of New Zealand albino rabbits and to study its effects in the epithelialization and inflammation of the wounds at 7 and 28 days after its application. STUDY DESIGN: A prospective study carried out on 20 adult rabbits. Two wounds were made on the midline of the dorsal surface of the tongue in each animal, one control, and the other in which PRGF was applied. A histological study of the epithelialization and inflammation of wounds at 7 and 28 days was made. RESULTS: At 7 days were not observed differences between the study group and the control, nevertheless at 28 days all the wounds in which we applied the PRGF were completely epithelialized and with resolution of the inflammatory process, finding significant differences with respect to the control (p=0.031) and (p=0.023). CONCLUSIONS: The PRGF accelerates epithelialization and reduces inflammation at 28 days of provoking wounds in the oral mucosa.

The use of plasma rich in growth factors on wound healing in the skin: experimental study in rabbits.

The objective of this study was to use autologous plasma rich in growth factors (PRGF) on wound healing in the skin in New Zealand albino rabbits and to study reepithelialisation and inflammation at 7 and 28 days. A prospective study carried out on 20 adult rabbits. Two wounds were made on the in the skin on the back of each animal; one control, and the other in which PRGF was applied. The PRGF preparation was obtained from 10 ml of whole blood. The reepithelialisation and inflammation of wounds were measured at 7 and 28 days. Reepithelialisation improved in skin at 7 days (P = 0.007), with resolution of the inflammatory process (P = 0.005), having significant differences with respect to the control. Therefore, PRGF accelerates reepithelialisation and reduces inflammation at 7 days in skin.

In vivo treatment of melanoma (B16F10) with a homeopathic agent and with a cytokine (IFN-alpha).

Among the numerous agents tested on melanoma, cytokines have attracted much attention over recent decades, in particular interferon-alpha (IFN-alpha). However, in a small number of experimental assays, homeopathic products have also been used. This study aimed to analyze the effects of INF-alpha and Lymphomyosot, administered individually or in combination, on the growth of B16F10 melanoma transplanted in C57BL/6J mice. Two experiments were performed using 72 young male mice, treated with 1 x 10(6) B16F10 cells and treated with phosphate-buffered saline (I), INF-alpha (II), Lymphomyosot (III), and both INF-alpha and Lymphomyosot (IV). Subsequent morphological and immunohistochemical studies were performed. All treatments produced a reduction in tumor weight with significant differences in those treated with INF-alpha and Lymphomyosot. INF-alpha reduced the cell proliferation index and the spread of inflammatory infiltrates and produced an increase in the extent of intratumoral necrosis. An antitumour effect was displayed by both agents, as was the cytotoxicity of INF-alpha and the immune response-stimulating effect of Lymphomyosot.

Radioprotective effects of lycopene and curcumin during local irradiation of parotid glands in Sprague Dawley rats.

Radiotherapy effectively treats cancers of the head and neck. We investigated the possible protective effects of lycopene and curcumin on the parotid glands of 40 female Sprague Dawley rats during irradiation. The study followed European Union regulations 86/609/EEC, 2010/63/EU for animal experimentation. The animals were divided into 4 groups: those treated with curcumin and radiation, those treated with lycopene and radiation, those treated with dimethyl sulphoxide (DMSO) and radiation, and those treated with radiation alone. All compounds were given intraperitoneally the day before irradiation. The total dose of radiation was 20Gy. Morphological and histopathological analyses showed less cell necrosis in the group treated with curcumin than in the other groups, but the difference was not significant. Analysis of structural damage to the parotid ducts and vacuolisation showed significant differences among all groups (p=0.023, p<0.01). Lycopene and curcumin given 24 hours before irradiation reduced the structural damage to the salivary glands. Further studies are needed to confirm these findings.

Treatment of metastatic melanoma B16F10 by the flavonoids tangeretin, rutin, and diosmin.

Melanoma is one of the most frequently metastasizing malignant neoplasias. This study examines an experimental model of pulmonary metastasis and the B16F10 cell subline, highly metastatic in the lung. Antimetastatic effects of the flavonoids tangeretin, rutin, and diosmin were analyzed, and at the same time an analysis of the metastatic activity of ethanol was performed, considered to be necessary because it is used as a vehicle for administering the flavonoids. Lentini's model, which complements the macroscopic evaluation of nodule numbers by using a stereoscopic microscope and image analysis at the microscopic level, was used. The greatest reduction in the number of metastatic nodules (52%) was obtained with diosmin; similarly, the percentages of implantation, growth index, and invasion index (79.40, 67.44, and 45.23%, respectively), were all compared with those of the ethanol group, considered to be an effective control group. Rutin- and tangeretin-treated groups also showed reductions of the same index compared with the ethanol group. It would seem that structural factors would better explain these results and the antimetastatic activity of each flavonoid and the respective metabolites.

[Experimental model for treating pulmonary metastatic melanoma using grape-seed extract, red wine and ethanol]. / Modelo experimental de tratamiento con extracto de semillas de uva, vino tinto y etanol del melanoma metastásico pulmonar.

INTRODUCTION: Melanoma is one of the neoplasias that most frequently metastasize, especially in the lung, where represents a challenge in oncology since current treatment is ineffective, and mortality is high. MATERIAL AND METHODS: Swiss mice (n = 52) were inoculated with 0.5 x 106 B16F10 cell lines and, later, given an oral administration of grape-seed extract, red wine or ethanol. Metastatic nodules on the lung surface were counted and, after processing for microscopy, five sections were selected for image analysis and the invasion index was calculated. RESULTS: Macroscopic analysis showed that grape-seed extract and red wine reduced the number of metastatic nodules by 26.07 and 20.81%, respectively, compared with a control group treated with ethanol. Microscopically, the reduction in the invasion index was 31.65 for grape-seed extract and 17.57% for red wine. CONCLUSION: Ethanol administration significantly increased pulmonary metastasis while grape-seed extract and red wine led to their reduction.

Influence of murine renal sexual dimorphism on amiloride-induced hyperkalemia.

The present study investigated the influence of androgens in potassium homeostasis both under normal conditions and when potassium balance was affected by the mild diuretic amiloride. Our results indicate the existence of a clear sexual dimorphism in renal and plasma potassium content in CD1 mice, adult males having a higher plasma potassium concentration than females or 40-day-old male mice (4.6 +/- 1.0 vs. 3.9 +/- 0.9 mEq/l). Plasma K(+) concentration was increased by testosterone treatment and decreased after orchiectomy, while the opposite occurred in the case of renal potassium concentration. Amiloride was well tolerated by CD1 female mice, but induced severe hyperkalemia in male mice, where it was also associated with a high rate of mortality. Testosterone treatment increased the toxicity of the diuretic in both CD1 female and male mice. Histopathological analysis of kidneys from CD1 male mice treated with amiloride revealed alterations at the proximal and distal tubule level. These results strongly suggest that androgens may produce adverse effects in renal function.

In-vivo behavior of Si-hydroxyapatite/polycaprolactone/DMB scaffolds fabricated by 3D printing.

Scaffolds made of polycaprolactone and nanocrystalline silicon-substituted hydroxyapatite have been fabricated by 3D printing rapid prototyping technique. To asses that the scaffolds fulfill the requirements to be considered for bone grafting applications, they were implanted in New Zealand rabbits. Histological and radiological studies have demonstrated that the scaffolds implanted in bone exhibited an excellent osteointegration without the interposition of fibrous tissue between bone and implants and without immune response after 4 months of implantation. In addition, we have evaluated the possibility of improving the scaffolds efficiency by incorporating demineralized bone matrix during the preparation by 3D printing. When demineralized bone matrix (DBM) is incorporated, the efficacy of the scaffolds is enhanced, as new bone formation occurs not only in the peripheral portions of the scaffolds but also within its pores after 4 months of implantation. This enhanced performance can be explained in terms of the osteoinductive properties of the DBM in the scaffolds, which have been assessed through the new bone tissue formation when the scaffolds are ectopically implanted.

Microsatellite pathologic score does not efficiently identify high microsatellite instability in colorectal serrated adenocarcinoma.

The microsatellite pathologic score has been proposed as a valuable tool to estimate the probability of a colorectal cancer having high microsatellite instability; however, this score has not been tested in serrated adenocarcinoma. Our aim was to evaluate microsatellite pathologic score in serrated adenocarcinoma, conventional carcinoma, and colorectal cancer with high microsatellite instability histologic features. Eighty-nine serrated adenocarcinoma and 81 matched conventional carcinomas were tested with microsatellite pathologic score, and the results were compared with those of 24 high microsatellite instability histologic features. Validation was performed by microsatellite instability analysis. Although all colorectal cancers with high microsatellite instability histologic features rendered a more than 5.5 score, the microsatellite pathologic score performance was of lower rank in high microsatellite instability serrated adenocarcinoma because none of the cases scored above 5.5 (>77% probability of being high microsatellite instability). High microsatellite instability serrated adenocarcinoma shows pathologic features different from those observed in high microsatellite instability histologic features such as adverse prognostic histologic features at the invasive front. We describe a serrated adenocarcinoma subtype showing high microsatellite instability and some, but not all, high microsatellite instability histologic features that would not be detected if the microsatellite pathologic score cutoff is set at the highest rank. To increase microsatellite pathologic score sensitivity in serrated adenocarcinoma, we propose to set up a 2.1 cutoff score when faced by a right-sided colorectal cancer with serrated features.

Plasma rich in growth factors and bone formation: a radiological and histomorphometric study in New Zealand rabbits.

A radiographic and histomorphometric study was conducted on the influence of autologous plasma rich in growth factors (PRGF) upon bone healing in surgically created defects in rabbits. Radiographically, bone regeneration was significantly greater with the use of PRGF after one month (p = 0.005), though no differences were recorded after the second month. In the histomorphometric analysis one month after surgery, the defects filled with autologous bone plus PRGF showed a greater percentage of neoformed bone (35.01 +/- 5.31) than the control defects (22.90 +/- 12.23), though the differences were not significant. Two months after surgery, the defects filled with autologous bone showed greater regeneration (46.04 +/- 10.36%) than the control defects (30.59 +/- 5.69%), though the differences were not significant. The application of PRGF in the bone defects produced in New Zealand rabbits exerted a limited effect on local bone formation.

In vitro and in vivo effect of IFNalpha on B16F10 melanoma in two models: subcutaneous (C57BL6J mice) and lung metastasis (Swiss mice).

Interferon alpha tends to be the only agent used to treat melanoma. The objective of this study was to assess the effect of interferon alpha on the growth of the B16F10 melanoma, both in vitro and in vivo. We study the in vitro effect of interferon alpha (250,000, 500,000 and 1,000,000 IU/ml) on the B16F10 melanoma cell line (at 24, 48 and 72 h) and the in vivo effect in a subcutaneous (1x10(6) cells; 300,000 IU) and a pulmonary metastatic model (5x10(5) cells/lateral vein of the tail; 300,000, 600,000 and 1,200,000 IU). Necropsy included a morphological and immunohistochemical study (subcutaneous model), while the number of superficial lung metastases, implantation percentage and growth and invasion indices were calculated in the latter model. In vitro, interferon alpha decreased cell survival in a time- and dose-dependent manner; 250,000 IU/ml: 77% (24h), 80% (48 h) and 92% (72 h); 500,000 IU/ml: 62% (24h), 32% (48 h), 20% (72 h); 1,000,000 IU/ml: 41% (24h), 16% (48 h), 10% (72 h). In the subcutaneous model, it reduced tumor weights (77.74%) and cell proliferation (70.8%), and increased necrotic areas (8%) and inflammatory infiltrates (34.46%). Metastatic model: 300,000 IU reduced pleural nodules by 38.79%, implantation by 59.42%, growth by 43.48%, invasion by 25.06%; the corresponding figures for 600,000 and 1,200,000 IU were 38.79, 59.42, 43.48, 25.06%, and 65.55, 84.98, 56.52, 66.19%, respectively. Interferon alpha inhibited cell proliferation in all the models and had immunomodulatory (subcutaneous model) and antimetastatic (pulmonary metastatic model) effects in vivo.

Oral and facial piercings: a case series and review of the literature.

INTRODUCTION: Piercing is popular among young people, who view this practice as a sign of marginality, beauty, or group identity. This study is performed on healthy individuals with oral and facial piercings. MATERIALS AND METHODS: Seventy oral and facial piercings were evaluated (17 in the tongue, 13 in the lower lip, 18 in the nostril, 7 in the eyebrow, and 15 in the ear). A specifically designed protocol was used to assess possible complications (inflammatory reactions, pain, dental alterations). Nonparametric tests were used for the statistical analysis of the results. RESULTS: The general complications recorded comprised pain (60% of cases), inflammatory reactions (34.3%), bleeding (24%), dental fractures or fissures (20%), and gingival damage (26.7%). CONCLUSION: Tongue piercing is associated with pain, inflammatory reactions, and dental problems.

Plasma rich in growth factors and bone formation: a radiological and histomorphometric study in New Zealand rabbits

A radiographic and histomorphometric study was conducted on the influence of autologous plasma rich in growth factors (PRGF) upon bone healing in surgically created defects in rabbits. Radiographically, bone regeneration was significantly greater with the use of PRGF after one month (p = 0.005), though no differences were recorded after the second month. In the histomorphometric analysis one month after surgery, the defects filled with autologous bone plus PRGF showed a greater percentage of neoformed bone (35.01 ± 5.31) than the control defects (22.90 ± 12.23), though the differences were not significant. Two months after surgery, the defects filled with autologous bone showed greater regeneration (46.04 ± 10.36 percent) than the control defects (30.59 ± 5.69 percent), though the differences were not significant. The application of PRGF in the bone defects produced in New Zealand rabbits exerted a limited effect on local bone formation.