Browsing by Subject "Histomorphometry"
- PublicationOpen AccessChanges in nitrosative stress biomarkers in swine intestine following dietary intervention with verbascoside(2013) Giancamillo, Alessia Di; Rossi, Raffaella; Vitari, Francesca; Carollo, Valentina; Deponti, Daniela; Corino, Carlo; Domeneghini, CinziaIn farm animals, oxidative stress can be involved in several intestinal pathological disorders, and many antioxidant molecules, especially those of plant origin, can counteract free radicals, thus stabilizing the gut environment and enhancing health. The aim of the study was to investigate whether the use of verbascoside (VB), a polyphenol plant compound, in pig feeding could modulate oxidative and/or nitrosative stress in the gut. Eighteen male piglets (Dalland) were assigned to two groups, which were fed with either a control diet (CON) or a diet supplemented with 5 mg/kg of verbascoside (VB) for 166 days. At slaughter, duodenum and jejunum specimens were collected. Immunohistochemistry and Western blot analyses were performed on the samples to evaluate free radical adducts, including acrolein (ACR), 8-hydroxydeoxyguanosine (8-OHdg) and nitrotyrosine (NT). A KRL test was also used to assess the total blood antioxidant activity, and no difference was observed. Immunohistochemistry and Western blot showed that dietary treatment decreased the levels of nitrotyrosine in enteroendocrine cell populations (P<0.05). Characterization of the enteroendocrine cell typology was then performed, and serotonin-immunoreactive cells were revealed to be directly involved in decreasing the nitrosative stress status. This preliminary study demonstrates the important role of dietary VB in decreasing stress biomarkers in swine gut, thus highlighting a possible intervention aimed at building a large prospective for antioxidant dietary supplementation in food animal species.
- PublicationOpen AccessDelayed remodeling in the early period of fracture healing in spontaneously diabetic BB/OK rats depending on the diabetic metabolic state(Murcia : F. Hernández, 2004) Follak, N.; Klöting, I.; Wolf, E.; Merk, H.Several clinical series, analyzing fracture healing in patients with insulin-dependent type 1 diabetes (IDDM) demonstrated significant incidence of delayed union, non-union, and pseudarthrosis. The purpose of this study was to examine the detailed histomorphometry and histology of bone formation and remodeling during fracture healing depending on the diabetic metabolic state in spontaneously diabetic BB/O(ttawa)K(arlsburg) rats, a rat strain that represents a close homology to IDDM in man. A standardized fracture model was chosen and based on blood-glucose values at the time of surgery (mg%), postoperative blood-glucose course (mg%) and postoperative insulin requirements (IU/kg), 100 spontaneously diabetic BB/OK rats were divided into groups with well-compensated (n=50, 167±77 mg%; 244±68 mg%; 1.8±1.9 IU/kg) or poorly compensated (n=50, 380±89 mg%; 415±80 mg%; 6.0±1.0 IU/kg) metabolic state. Fifty LEW.1A rats served as the normoglycemic controls (97±15 mg%). Ten animals from each group were killed 1, 2, 3, 4 and 6 weeks after fracture and specimens were processed undecalcified for quantitative histomorphometry and for qualitative light microscopy. In terms of bone histomorphometry, within the first four weeks after fracture, severe mineralization disorders occurred exclusively in the rats with poorly compensated diabetic metabolic states with a significantly decrease of all fluorochrome-based parameters of mineralization, apposition, formation and timing of mineralization in comparison to the spontaneously diabetic rats with wellcompensated metabolic states and to the control rats. This was confirmed histologically. Early fracture healing in the spontaneously diabetic BB/OK rats is delayed exclusively in poorly compensated diabetic metabolic states, and 6 weeks after fracture, histomorphometrically significant deficits in the measured and dynamically calculated parameters remain. This study suggests that strictly controlled insulin treatment resulting in well-compensated diabetic metabolic states will ameliorate the impaired early mineralization and cell differentiation disorders of IDDM fracture healing.
- PublicationOpen AccessDemineralized bone matrix coating Si-Ca-P Ceramic does not improve the osseointegration of the scaffold(MDPI, 2018-09-01) Parrilla-Almansa, Andrés; García Carrillo, Nuria; Ros-Tárraga, Patricia; Martínez, Carlos Manuel; Martínez Martínez, Francisco; Meseguer Olmo, Luis; De Aza, Piedad N.; Cirugía, Pediatría y Obstetricia y GinecologíaThe aim of this study was to manufacture and evaluate the effect of a biphasic calcium silicophosphate (CSP) scaffold ceramic, coated with a natural demineralized bone matrix (DBM), to evaluate the efficiency of this novel ceramic material in bone regeneration. The DBM-coated CSP ceramic was made by coating a CSP scaffold with gel DBM, produced by the partial sintering of different-sized porous granules. These scaffolds were used to reconstruct defects in rabbit tibiae, where CSP scaffolds acted as the control material. Micro-CT and histological analyses were performed to evaluate new bone formation at 1, 3, and 5 months post-surgery. The present research results showed a correlation among the data obtained by micro-CT and the histomorphological results, the gradual disintegration of the biomaterial, and the presence of free scaffold fragments dispersed inside the medullary cavity occupied by hematopoietic bone marrow over the 5-month study period. No difference was found between the DBM-coated and uncoated implants. The new bone tissue inside the implants increased with implantation time. Slightly less new bone formation was observed in the DBM-coated samples, but it was not statistically significant. Both the DBM-coated and the CSP scaffolds gave excellent bone tissue responses and good osteoconductivity.
- PublicationOpen AccessEffect of N-Ethyl-N-Butyl-Nitrosamine on the esophageal mucosa of the rat. Histometric investigation of early tumor stages(Murcia : F. Hernández, 1987) Sons, H.U.; Borchard, F.; Kaup, F.G.80 male Wistar rats received an oral ad libitum application of N-ethyl-N-butyl-nitrosamine in a concentration of 0.18 g per litre of drinking water. The changes induced in the esophakeal mucosa and determined at three intervals (up to 48 days, up to 91 days, and up to 112 days after commencement of carcinogen exposure) were compared by microscopy with the results from a control group of 10 male Wistar rats of the same age. Several histomorphometric parameters were investigated with the aid of a Leitz ocular micrometer. The earliest localized changes found were an increase in the thickness of the epithelium and the horny layer, and an elongation of the papillary bodies and a widening of the parabasal cellular layer. Later - with a substantial increase in the rate of mitosis in all layers of the epithelium, there was a significant thickening of the non-papillomatous and papillomatous epithelium, an enlargement of the nuclei, especialli in the middle and upper layers of the epithelium and a thickening of the horny layer, parts of the latter being parakeratotic. The papillomatous changes corresponded in some cases to moderate epithelia1 dysplasias. As expected, no fully-developed invasive carcinomas occurred in the early period investigated. The histometric data permit the conclusion to be drawn that the lesions described are demonstrable not only at the exophytic-papillomatous epithelium but also in multifocally localized form at the flat, nonpapillomatous mucosa, and that they can definetely be regarded as the expression of an incipient field cancerification
- PublicationOpen AccessHistomorphometric study of femoral heads in hip osteoarthritis and osteoporosis(Murcia : F. Hernández, 2007) De Pedro, J.A.; Martin, A.P.; Blanco, J.F.; Salvado, M.; Perez, M.A.; Cardoso, A.; Collía, F.; Ellis, S.S.; Domínguez, J.During the period from 2000 to 2003, ninety eight samples of femoral heads were collected. In these pieces, two zones were analyzed: a high-load zone (the hard core of the head) and a low-load zone (the round ligamentum teres zone). As control group, 6 femoral heads (3 of women and 3 of men), proceeding from autopsy in peoples without pathological antecedents and youngs, were studied. After the samples had been embedded in methylmethacrylate and stained, they were subjected to an histomorphometric study. By means of histomorphometry, trabecular bone volume (TBV) and osteoid substance (OSV) was determined. Statistically significant differences were found as for peripheral osteoid volume (low-load zone) ( p=0.036) and trabecular bone volume, both peripheral and central. Both volumes decreased in osteoporotic samples and in those from women (p=0.000), in comparison with control group. Regarding the relationship between the high-load and low-load zone, significant data were obtained. The high-load zone had a greater trabecular bone volume than the low-load zone, regardless of the pathology and sex, but this increase was more pronounced in the arthrosic samples and in those from men. Additionally, this trabecular bone volume in the high-load zone decreased with increasing age of the donor (p=0.037), when the control group is compared. In sum, we observed a reduction in the formation of TBV and OSV in osteoporosis but also a decrease in the arthrosic, in samples from older subjects, in women, and in the low-load zone of the samples, when the control group is compared. These data suggest the coexistence of both pathologies, which is more pronounced in older subjects and women.
- PublicationOpen AccessLow-intensity pulsed ultrasound increased blood vessel size during fracture healing in patients with a delayed-union of the osteotomized fibula(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Korstjens, Clara M.; Rutten, Sjoerd; Nolte, Peter A.; van Duin, Marion A.; Klein Nulend, JennekeDisturbed vascularity leads to impaired fracture healing. Since low-intensity pulsed ultrasound (LIPUS) increases new bone formation in delayedunions, we investigated whether LIPUS increases blood supply in delayed-unions of the osteotomized fibula, and if LIPUS-increased bone formation is correlated to increased blood supply. Blood vessel parameters were analysed using histology, immunohistochemistry, and histomorphometric analysis as well as their correlation with bone formation and resorption parameters. Fibular biopsies of thirteen patients with a delayed-union of the osteotomized fibula treated for 2-4 months with or without LIPUS originating from a randomized prospective double-blind placebo-controlled clinical trial were studied. In histological sections of the fibular biopsies parameters of blood vessel formation were measured and were related to histomorphometric bone characteristics of newly formed bone of the same samples analysed in our previously published study on the effects of LIPUS on bone healing at the tissue level in delayed-unions. LIPUS-treated delayed-unions and sham-treated delayed-unions as well as healed delayedunions and failed-to-heal delayed-unions were compared. The volume density of blood vessels was increased in LIPUS-treated delayed-unions compared to sham-treated controls. LIPUS did not change blood vessel number, but significantly increased blood vessel size. Healed delayed-unions as well as LIPUS-treated and sham-treated delayed-unions showed significant correlations between blood vessel size and osteoid volume. LIPUS increases blood vessel size, essential for fracture healing, in bone from patients with a delayedunion of the osteotomized fibula. The increased osteoid volume in delayed-unions can largely be explained by increased blood supply and perfusion.
- PublicationOpen AccessMicroscopic study of chronic Charcot arthropathy foot bones contributes to understanding pathogenesis - a preliminary report.(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Dharmadas, Salini; Kumar, Harish; Pillay, Minnie; Jojo, Annie; Tessy, P.J.; Mangalanandan, Thacho Sukumaran; Vivek, Lakshmanan; Praveen, Valiyaparambil Pavithran; Bal, ArunIntroduction. Charcot arthropathy (CA) is non-infective, chronic destructive condition affecting the pes architecture of long standing diabetic patients with neuropathy. Even though several theories have emerged to disclose its pathogenesis, inflammatory cytokine induced osteoclastogenesis stands as the chief culprit. Studies on micro-architecture of foot bones of acute stage CA patients, describes mainly destructive phase of bone remodelling. Increased osteoclast cell activity is reported in all studies communicated. No study has to the best of our knowledge detailed the microscopic structure of chronic stage CA foot bones. Aim. To study the microscopic structure of foot bones in patients with chronic CA. Materials and methods. Foot bones were collected from the feet of chronic CA patients (six in number) who underwent corrective foot surgery in the department of Podiatric Surgery of a tertiary care hospital. Control samples were collected from the feet of age matched non-diabetic controls (2 in number). The samples were fixed in formalin, decalcified in 10% nitric acid, processed, sectioned and stained with haematoxylin and eosin. Histopathology and histomorphometry analysis were performed by two different pathologists. Results. Trabeculae of chronic CA foot bones exhibited mainly a lamellar architecture, with reduced number of osteocytes and plenty of empty lacunae. Trabecular connectivity was lost and trabeculae showed considerable thinning. Trabecular osteoids lined by active osteoblast cells was a remarkable observation. Bone area was also considerably reduced in chronic CA foot bones. Conclusion. Chronic stage CA foot bones presented features of both healing and fragile bone. The compromised bone quality may be due to thin and fragmented trabecular structure and reduced cellularity.